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Hindawi Publishing CorporationClinical and Developmental Immunology Volume 983090983088983089983091 Article ID 983097983096983089983092983094983096 983097 pageshttpdxdoiorg983089983088983089983089983093983093983090983088983089983091983097983096983089983092983094983096
Clinical Study Cysticerci Drive Dendritic Cells to Promote In Vitro andIn Vivo Tregs Differentiation
Laura Adalid-Peralta12 Asiel Arce-Sillas2 Gladis Fragoso3 Graciela Caacuterdenas1 Marcos
Rosetti3 Didier Casanova-Hernaacutendez2 Claudia Rangel-Escarentildeo4 Laura Uribe-Figueroa5
Agnes Fleury123 and Edda Sciutto23
983089 Instituto Nacional de Neurolog ıa y Neurocirug ıa Insurgentes Sur 983091983096983095983095 Col La Fama 983089983092983090983094983097 M exico DF Mexico983090
Unidad Perif erica para el Estudio de Neuroin1047298amacion del Instituto de Investigaciones Biomedicas de la UNAM en el InstitutoNacional de Neurolog ıa Neurocirug ıa Insurgentes Sur 983091983096983095983095 Col La Fama 983089983092983090983094983097 M exico DF Mexico
983091 Instituto de Investigaciones Biomedicas Universidad Nacional Aut onoma de M exico 983088983092983093983089983088 M exico DF Mexico983092 Departamento de Genomica Computacional Instituto Nacional de Medicina Genomica INMEGEN Perif erico Sur 983092983096983088983097 Arenal Tepepan 983089983092983094983089983088 M exico DF Mexico
983093 Unidad de Genotipi1047297cacion y Analisis de Expresion Affymetrix INMEGEN Perif erico Sur 983092983096983088983097 Arenal Tepepan 983089983092983094983089983088 M exicoDF Mexico
Correspondence should be addressed to Laura Adalid-Peralta lauraadalidgmailcom
Received 983089 March 983090983088983089983091 Accepted 983090983092 April 983090983088983089983091
Academic Editor Arnon Nagler
Copyright copy 983090983088983089983091 Laura Adalid-Peralta et al Tis is an open access article distributed under the Creative Commons Attribution
License which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited
Regulatory cells (regs) play a crucial role in immune homeostasis reg induction is a strategy that parasites have evolvedto modulate the hostrsquos in1047298ammatory environment acilitating their establishment and permanence In human Taenia soliumneurocysticercosis (NC) the concurrence o increased peripheral and central reg levels and their capacity to inhibit cellactivation and prolieration support their role in controlling neuroin1047298ammation Tis study is aimed at identi1047297ng possiblemechanisms o reg induction in human NC Monocyte-derived dendritic cells (DC) rom healthy human donors cocultivated
with autologous CD983092+ naıve cells either in the presence or absence o cysticerci promoted CD983090983093highFoxp983091+ reg differentiation Anincreased reg induction was observed when cysticerci were present Moreover an augmentation o suppressive-related molecules(SLAMF983089 B983095-H983089 andCD983090983088983093) was ound in parasite-inducedDC differentiation Increasedregs and a higherinvivo DC expressiono the regulatory molecules SLAMF983089 and CD983090983088983093 in NC patients were also ound SLAMF983089 gene was downregulated in NC patientswith extraparenchymal cysticerci exhibiting higher in1047298ammation levels than patients with parenchymal parasites Our 1047297ndingssuggest that cysticerci may modulate DC to avor a suppressive environment which may help parasite establishment minimizing
the excessive in1047298ammation which may lead to tissue damage
1 Introduction
Natural (thymic) and inducible regulatory cells (regs)play a pivotal role in maintaining the immune system home-ostasis While natural regs are produced in the thymus atany time inducible regs acquire a regulatory unction inthe context o a given inection or a neoplastic process [983089]A variety o inducible reg subpopulations mediating theirimmune suppressive effects by different mechanisms havebeen reported [983090ndash983092] Disregarding their effectiveness in
controlling in1047298ammation it is conceivable that regs couldpromote a more permissive environment or parasite estab-lishment [983093 983094] Te increased reg levels ound in many different protozoa and cestode inections are consistent withthis possibility [983095 983096]
Te role o regs in neurocysticercosis (NC) a para-sitic disease caused by the establishment o Taenia soliummetacestode in the central nervous system (CNS) beginsto be explored Increased central and peripheral reg levelswere observed in severe NC patients Tese regs seem to
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983090 Clinical and Developmental Immunology
participate in the control o the in1047298ammatory response sincea negative correlation between the percentage o peripheralregs and activated CD983096+ and CD4+ cells along with adepressed cell prolierative response was observed [983097]However the mechanisms underlying reg induction in NCare still unknown
Dendritic cells play a prominent role in reg inductionin the beginning o the immune response to pathogenseither by promoting the conversion o naıve cells to regsubpopulationsor by expanding the population o preexistingreg cells [983089983088] Mature DCs direct conventional CD983092+ cells tobecome either speci1047297c helper or regulatory cell subsetsdepending on the affinity o their CR to the antigen thestrength o the costimulatory signals provided by antigen-presenting cells (APCs) and the cytokine milieu In additionthe absence o in1047298ammation arrests dendritic cells into animmature or semimature state (iDC) which promotes cell tolerance by conversion o naıve cells into regs [983089983089]iDCs are characterized by expressing MHC II by a highphagocytosis capacity and by low CD983096983088CD983096983094 expressioniDCs also produce IL-983089983088 but neither IL-983089983090 nor NF [983089983088983089983090] Moreover it has been proposed that preexisting regscan educate iDCs to become tolerogenic promoting reggeneration [983089983088 983089983091] Helminths as well as other pathogensmay modulate the immune tolerance properties o DCs [983089983092ndash983089983095]
Tis study was designed to evaluate the capacity o cysticerci to modulate dendritic cells driving them to reginduction both in vitro and in vivo
2 Materials and Methods
983090983089 Parasites Taenia solium metacestodes were obtainedrom naturally inected pigs coming rom villages o Guer-rero an endemic region in Mexico Pigs were euthanizedaccording to ethical animal handling regulations in MexicoCysticerci wereindividually harvested rom muscle tissue andmaintained in RPMI 983089983094983092983088 medium containing 983089983088 etal cal serum and 983089 antibiotics (Invitrogen NY USA) Cysticercio Taenia crassiceps ORF strain were also used in this study to evaluate their potential to induce iDCs and thereoreregs T crassiceps cysticerci were obtained rom inectedBALBcAnN emale mice Parasites were harvested rom theperitoneal cavity o stock emale mice afer 983089983088 weeks o inection
983090983090 Cell Puri1047297cationand DC Generation in thePresenceof LiveCysticerci Human cells were isolated rom buffy coats romblood o healthy donors (Banco de Sangre del Centro MedicoNacional Siglo XXI Mexico DF)
A RosetteSep Human CD983092+ Cell Enrichment Cocktailkit was used or CD983092+ cell puri1047297cation (StemCell Vancou-
ver Canada) Puri1047297ed CD983092+ lymphocytes including gt983097983088o CD983091+ CD983092+ cells were kept rozen at ndash983096983088∘C until used
Monocyte-derived DCs (MDDCs) were isolated rommononuclear cells rom healthy donors using a Rosette-Sep Human Monocyte Enrichment Cocktail kit (StemCellVancouver Canada) MDDCs were generated as previously
reported [983089983096] with only minor modi1047297cations Brie1047298y mono-cytes were plated in Petri dishes (P983089983088983088) at 983093 times 9830899830885 cellsmLin 983089983088 mL o RPMI 983089983094983092983088 medium (Invitrogen NY USA)containing 983089983088 human AB serum (HSAB) and 983089 antibiotics983090983088 ngmL IL-983092 (eBiosciences CA USA) and 983089983088983088 ngmLgranulocyte-macrophage colony-stimulating actor (eBio-
sciences San Diego CA USA) or 983096 days One hal o complete medium including cytokines was replaced at day 983092
o evaluate the role o cysticerci during DC differenti-ation monocytes were cultured as described above eitherwith or without vesicular Taenia crassiceps (983092983088 cysts) orTaenia solium (983093 cysts) cysticerci per dish Te effect o both cestodes on DC differentiation was tested consideringthat murine cysticerci may eventually provide a controlledparasite source o puri1047297ed immune-modulatory componentsDexamethasone at 983089983088minus7 M was used as a positive control
983090983091 In Vitro Treg Induction with Cysticercal Antigens Para-
site-induced MDDCs were seeded at 983090983093 times 9830899830884
cellmL in983089 mL RPMI 983089983094983092983088 medium (Invitrogen NY USA) containing983089983088 human AB serum and 983089 antibiotics (Invitrogen NYUSA) MDDCs were maintained in culture or 983090983092 h Dur-ing this period dexamethasone at 983089983088minus7 M which maintainsa tolerogenic DC phenotype was used as control [983089983097]Parasite-induced DCs were pulsed using 983091 1038389gmL o totalantigens rom T crassiceps cysticerci T crassiceps antigenswere employed considering that T solium and T crassicepscysticerci share in common more than 983097983096 o the expressedantigens [983090983088] 983089983090983093 times 9830899830885 autologous CD983092+ lymphocyteswere cocultured or 983095 days with MDDCs in RPMI 983089983094983092983088medium (Invitrogen NY USA) containing 983089983088 human AB
serum and 983089 antibiotics (Invitrogen NY USA) Tenthe phenotype o regulatory cells was evaluated by 1047298ow cytometry as described below Supernatants o these cultureswere used to evaluate cytokine pro1047297le
983090983092 Cell Phenotype Te ollowing combination o antibodieswasused to characterizethe phenotype o MDDCs(a) MHC-II FIC (mouse IgG983090a k) SLAMF983089 PE (mouse IgG983089 k) CD983089983089cPerCP-eFluor 983095983089983088 (mouse IgG983089 k) and B983095H983089 APC (mouseIgG983089 k) (b) CD983096983091 FIC CD983096983088 PE CD983089983089c PerCP-eFluor 983095983089983088(mouse IgG983089 k) and CD983096983094 APC (mouse IgG983089 k) (c) CD983090983088983093FIC (mouse IgG983090b k) CD983092983088 PE (mouse IgG983089 k) CD983089983089cPerCP-eFluor 983095983089983088 (mouse IgG983089 k) and IL983091 APC (mouse
IgG983089 k) Inducible regulatory cell phenotype was assessedusing CD983089983090983095 FIC (mouse IgG983089 k) Foxp983091 PE (Rat IgG983090a k)CD983092 PerCp (mouse IgG983089 k) and CD983090983093 APC (mouse IgG983089 k)Regulatory cells were intracellularlystained orFoxp983091usingthe eBiosciences kit (most antibodies used or cell stainingwere purchased rom eBiosciences CA USA) All antibodieswere titrated or optimal detection o positive populationspriorto its use considering the manuacturerrsquos recommendedconcentrations
For cytometry analyses cells were 1047297rst gated according tolymphocyte orward and sidelight-scattering propertiesregnumbers were de1047297ned as the raction o Foxp983091+ cells among
CD983092+ CD983090983093high cells
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Clinical and Developmental Immunology 983091
983090983093 Cytokines Cytokine levels in supernatants were mea-sured using the cytometric bead array Cytokine Kit II(BD Biosciences Pharmingen CA USA) according to themanuacturerrsquos instructions in an FACSCalibur cytometerData were analyzed with the BD Cytometric Bead Array sofware (BD Biosciences Pharmingen CA USA) Flow
cytometer was calibrated using BD FACSComp (BD Bio-sciences Pharmingen CA USA) and BD CaliBRIE beads(BD Biosciences Pharmingen CA USA) Assay sensitivitieswere set as ollows IL-983090 (983090983094 pgmL) IL-983094 (983091983088 pgmL) IL-983089983088 (983090983096 pgmL) NF- (983090983096 pgmL) and IFN-1103925 (983095983089 pgmL)GF-907317 were measured using the humanmouse GF-beta983089 ELISA Ready-SE-Go (eBiosciences CA USA) ELISAwas perormed according to the manuacturerrsquos instructionsdetection limit was 983094983088 pgmL All samples were run induplicate
983090983094 DC and Treg Phenotype in NC Patients Blood samplesrom 983089983091 patients who attended at the Instituto Nacional deNeurologıa y Neurocirugıa and Centro Medico NacionalSiglo XXI in Mexico City with con1047297rmed NC diagnosis wereincluded in this study All samples were taken beore any treatment was administered Eight male (age mean 983093983094983090983093 plusmn983091983096 years) and 1047297ve emale (age mean 983091983095 plusmn 983089983094 years) patientswere included in this study
In all cases NC was diagnosed based on clinical manies-tations (seizures ocal de1047297cit and intracranial hypertension)and radiological studies such as MRI and computed tomog-raphy (C)
Vesicular parasites were observed in most cases (983089983090983089983091patients) A solitary cysticercus was seen in 983097 patients andmultiple cysticerci in 983090 patients the rest o the patients
had calci1047297ed and colloidal orms In seven patients par-asite was located at the subarachnoid space o the base(SAB) while the rest o the patients showed parasites atthe cerebral parenchyma (P) Patients with SAB cysticerciexhibited in1047298ammatory traits in cerebral spinal 1047298uid (glucose983091983088983095 plusmn 983090983096 mgdL proteins 983092983090983089 plusmn 983092983088983095 mgdL and 983095983088983095 plusmn
983097983094983097 cellmm3) In contrast patients with parenchymal cys-ticerci showed no in1047298ammation signs Blood samples rom 983093healthy subjects were included as controls
983090983095 Microarrays DNA microarrays were used to comparethe gene expression pro1047297le o mononuclear peripheral cells
(PBMCs) rom a cohort o 983096 NC patients 983092983088 plusmn 983089983092 years oldPBMC rom all patients were obtained 983096 days afer cysticidaltreatment with albendazole 983091983088 mg per kg and concomi-tant corticosteroid administration Tese 983096 (983092 emales and983092 males) patients showed cysticerci in different locationsCysticerci were establishedin the subarachnoid basal space orin the ventricles (SABIV) in 983093 patients and were establishedin the cerebral parenchyma (P) in 983091 patients When requiredor patientsrsquo ollowup CSF was obtained by lumbar punctureat the National Institute o Neurology and Neurosurgery o
Mexico City A mean o 983095983092 cellmm3 (range 983089ndash983090983094983089) was ound
in SABIV-NC patients and a mean o 983094983094 cellmm3 (range983088ndash983089983096) cells was ound in P-NC patients PBMCs were isolated
and then cultured by 983095983090 h with T solium cysticercal antigensobtained as previously described [983090983089]
Recovered cells were placed in RIzol (Gibco NY USA)or RNA puri1047297cation using the RNeasy mini kit (QIAGENX USA) ollowing the manuacturerrsquos instructions Teamount o obtained RNA was estimated in an Agilent 983090983089983088983088
Bioanalyzer (Agilent echnologies Waldbronn Deutsch-land) Comparisons were done with respect to parasitelocation (SABIV-NC versus P-NC) using the R-languageplatorm under the terms o the Free Sofware FoundationsGNU General License (see httpcranr-projectorg) Datawere standardized and normalized All genes were analyzedusing the Reactome sofware (see httpwwwreactomeorg)to 1047297nd a possible biological pathway involved
983090983096 Ethical Considerations Te present study ul1047297lled allregulations or research with human subjects as requiredby the Mexican law and international regulations It alsocomplied with all ethical aspects considered in the General
Rules o Health or Clinical Investigation Ethics Commit-tee at Instituto Nacional de Neurologıa y NeurocirugıaMexico approved the protocol Written inormed consent wasobtained rom all participants Patients were inormed thatsamples obtained would be used or this work
983090983097 Statistical Analysis Data were processed in InStat(GraphPad Sofware Inc CA USA) Variables were describ-ed using mean plusmn SD Differences between groups were cal-culated with Studentsrsquo -test values less than 983088983088983093 wereconsidered signi1047297cant
3 Results
983091983089 Effect of Cysticerci on Dendritic Cell Phenotype Te semi-mature DC phenotype is associated with tolerance and reginduction [983089983090] o evaluate the effect o cysticerci on dendriticcell differentiation the phenotype o DC generated romCD983089983092+ monocytes eitherin thepresence or absence o vesicu-lar cysticerci afer 983096-day culture was studied Dexamethasone(983089983088minus7 M) was employed as a positive control (Figure 983089) Teexpression o CD983096983091+ as well as the costimulatory (CD983096983088CD983096983094 CD983092983088 HLA-DR) and regulatory molecules (SLAMF983089B983095-H983089 CD983090983088983093 and IL983091) on CD983089983089c+ was measured Asshown in Figure 983089 a higher expression o SLAMF983089 B983095-
H983089 and CD983090983088983093 was observed in cells differentiated in thepresence o cysticerci Te expression o HLA-DR CD983096983088CD983096983091 CD983096983094 CD983092983088 and IL983091 molecules did not differbetween control and parasite-driven differentiated DC cellsOn the other hand CD983096983088 CD983092983088 and SLAMF983089 were dimin-ished in dexamethasone-treated cells with respect to control(Figure 983089) HLA-DR and IL983091 expression did not differ romcontrol at any tested condition (data not shown) Altogetherthese results indicate that parasite promotes the expression o molecules related to a DC tolerogenic phenotype
983091983090 Taenia Solium and T crassiceps Cysticerci Promote In VitroRegulatory T Cells Differentiation We investigated whether
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983092 Clinical and Developmental Immunology
Medium Dex
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4053 3155 5606
1351 1393 1839
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484 028 129
5380 2739 8256
7455 9987 9933
4733 5947 6413
DC differentiated in the presence of
C D 8 0
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T crassiceps cysticerci
F983145983143983157983154983141 983089 Parasite effect on DC differentiation and activation Monocytes were differentiated to DC in presence o IL-983092 and GM-CSF plusmedium or dexamethasone or Taenia crassicepscysticerci On day 983096 the DC phenotype wasstudiedRepresentative data o three independentexperiments are shown Expression o CD983096983088 CD983096983091 CD983096983094 CD983092983088 SLAMF983089 B983095-H983089 andCD983090983088983093 gated on CD983089983089c+ cells was analyzed by FACSTe isotype controls are shown in white histograms
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Clinical and Developmental Immunology 983093
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208 29 426 344
FOXP3
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F983145983143983157983154983141 983090 In vitro differentiation o dendritic cells in the presence o Taenia solium or T crassiceps cysticerci promotes reg induction (a)Increased percentage in reg cell induction Percentage was calculated as ollows (percentage o regs inducedpercentage o basal regs)times 983089983088983088 Different letters indicate signi1047297cant differences between groups at lt 005 (b) Representative histograms showing Foxp983091 inductionwithin CD983092+ CD983090983093high cells Data are representative o three independent experiments
differentiation in vitro in the presence o live cysticerci stim-
ulated the differentiation to CD983092+ CD983090983093high Foxp983091+ regsHuman DCs either differentiated in the presence or absenceo cysticerci were pulsed or not with cysticercal proteins andused to stimulate autologous CD983092+ lymphocytes Seven
days later the expression o CD983090983093high and Foxp983091 withinCD983092+ lymphocytes was measured As a positive controlo reg induction differentiated DCs were treated withdexamethasone (983089983088minus7 M) Asshown in Figure 983090 cocultivationwith cysticerci signi1047297cantly increased the percentage o CD983092+
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983094 Clinical and Developmental Immunology
983137983138983148983141 983089 Levels o cytokines in supernatants rom in vitro DC differentiation and in vitro regulatory cell induction
IFN-1103925 NF- IL-983089983088 IL-983094 IL-983090 GF-907317
DC nonstimulated 983088983091983097 plusmn 983088983096983096 983091983097983096 plusmn 983091983096983090 983091983089983095 plusmn 983089983095983097 gt983093983088983088983088 983088983091983092 plusmn 983088983095983095 983092983088983094983093983088 plusmn 983089983097983089983094983097daggerControl medium 983089983088 HSAB 983091983093983093 plusmn 983088983093983089 983093983095983096 plusmn 983089983095983097 983093983089983090 plusmn 983089983093983088 gt983093983088983088983088 983089983093983097 plusmn 983089983092983089 NDb
DC differentiation in the presence o
Dexamethasone 983088 983091983089983090 plusmn 983088983092983095 983091983088983096983092 plusmn 983093983096983090a
gt983093983088983088983088 983088 983093983092983093983089983095 plusmn 983090983096983097983091983095Taenia crassiceps 983088983095983096 plusmn 983089983095983093 983090983097983096 plusmn 983089983095983093 983091983095983095 plusmn 983089983095983094 gt983093983088983088983088 983088983095983090 plusmn 983088983097983096 983091983092983089983093983088 plusmn 983090983091983096983089983097
Taenia solium 983088 983092983093983096 plusmn 983092983092983092 983095983091983089 plusmn 983092983096983096 gt983093983088983088983088 983088 983092983091983094983088983088 plusmn 983090983092983091983097983093
reg induction rom naıve CD983092+ cells
DC nonstimulated nonpulsed (Agminus) 983088983095983094 plusmn 983089983091983089 983089983090983089 plusmn 983089983088983095 983088983097983097 plusmn 983088983097983091 983090983093983096 plusmn 983088983095983088 983088983092983093 plusmn 983088983095983096 983089983090983092983094983088 plusmn 983093983091983093
DC nonstimulated pulsed (Ag+) 983088983095983096 plusmn 983089983095983093 983090983088983094 plusmn 983088983096983096 983089983090983090 plusmn 983089983089983090 983090983095983096 plusmn 983088983094983088 983088983092983091 plusmn 983088983097983093 983097983091983088983095 plusmn 983089983094983097983096
Dexamethasone nonpulsed (Agminus) 983088983094983091 plusmn 983089983090983094 983089983094983096 plusmn 983088983091983092 983089983088983089 plusmn 983089983089983096 983090983097983095 plusmn 983090983088983090 983088983091983097 plusmn 983088983095983096 983097983093983092983097 plusmn 983089983097983094983094
Dexamethasone pulsed (Ag+) 983089983088983088 plusmn 983089983095983092 983090983092983097 plusmn 983088983091983094 983090983091983090 plusmn 983089983091983094 983090983095983096 plusmn 983089983089983088 983089983091983096 plusmn 983089983090983095 983097983096983094983091 plusmn 983090983094983091983097
T crassiceps nonpulsed (Agminus) 983089983090983091 plusmn 983090983089983091 983090983088983097 plusmn 983088983095983097 983089983088983090 plusmn 983089983095983094 983096983089983093 plusmn 983089983090983094 983089983091983089 plusmn 983089983090983089 983097983097983090983090 plusmn 983091983089983094983092
T crassiceps pulsed (Ag+) 983089983090983094 plusmn 983089983096983089 983090983089983096 plusmn 983088983091983096 983089983092983092 plusmn 983088983096983089 983090983097983091983092 plusmn 983092983094983095983093 983088983092983091 plusmn 983088983097983093 983089983089983097983096983088 plusmn 983092983090983093983094
T solium nonpulsed (Agminus) 983092983093983088 plusmn 983090983094983089 983090983091983093 plusmn 983088983094983096 983089983092983096 plusmn 983090983089983088 983093983094983097983097 plusmn 983093983090983089 983089983090983089983092 plusmn 983089983089983095983097 983089983089983089983088983088 plusmn 983091983093983092
T solium pulsed (Ag+) 983089983095983091 plusmn 983090983092983093 983090983090983096 plusmn 983088983094983096 983089983097983093 plusmn 983088983088983097 983089983089983094983096 plusmn 983094983088983090 983094983094983089 plusmn 983094983089983094 983095983096983093983088 plusmn 983090983089983090983089
Control cell 983089983093983092 plusmn 983089983092983089 983089983097983088 plusmn 983088983092983096 983089983095983095 plusmn 983088983097983097 983090983088983089 plusmn 983088983092983093 983088983096983096 plusmn 983089983090983088 983089983088983089983088983088 plusmn 983089983094983093983096aSigni1047297cantly different ( lt 005) compared to nonstimulated DC b
Human AB sera rom healthy donors were used in DC differentiation experimentsdaggerLevels o cytokines in the control medium supplemented with 983089983088 HSAB are shown
lymphocytes coexpressing the CD983090983093high and Foxp983091 ractionwith respect to non-stimulated DC rom 983089983089983093 plusmn 983089983091983095 to 983090983089983092983094 plusmn983095983094983091when cocultured with T crassiceps ( = 004)andto983091983090983089983090plusmn 983089983089983093 ( = 005) when cocultured with T solium cysticerciWhen DCs were pulsed with parasite antigens regcells werealso signi1047297cantly increased rom 983089983088983089983090 plusmn 983091983091 to 983090983095983091 plusmn 983096983094983093( = 001) with T crassiceps-differentiated DC cells and to983091983091983097983091 plusmn 983091983096983094 ( = 00002) with T solium-differentiated DCcells (Figure 983090)
983091983091 Cytokine Pro1047297le Te levels o induced cytokines weremeasured in supernatants rom in vitro DC differentiationafer 983096 days o culture and rom in vitro reg inductionafer 983095 days o culture During DC differentiation only IL-983089983088 was signi1047297cantly increased when cells were differentiatedin presence o dexamethasone (able 983089) No difference wasobserved in the other tested conditions
983091983092 Dendritic Cells and Regulatory T Cells in NC PatientsTe in vivo effect o parasite products on the phenotype o peripheral DC rom NC patients was studied Te percentage
o DC andthe expression o regulatory(SLAMF983089 CD983090983088983093 andIL983091) and costimulatory molecules (HLA-DR CD983096983094 andCD983092983088) in CD983089983089c cells in 983089983091 NC patients and 983093 healthy subjectswas measured As able 983090 shows SLAMF983089 and CD983090983088983093 aresigni1047297cantly increased in DC rom NC patients rom 983091983096 to983097983095 and rom 983092983091 to 983090983089983093 respectively ( lt 005) Moreoverperipheral regs are also increased in NC patients rom 983092983091983093to 983089983092983090983090
983091983093 SLAMF983089 Is Downregulated in PBMC from Patients withSABIV Parasites As able 983091 shows only our immune-related genes o the 983091983090983091983090983090 included in the array were ounddownregulated in severe SABIV-NC patients with respect to
983137983138983148983141 983090 Phenotype o peripheral dendritic and regulatory cells inNC patients and healthy subjects
Phenotype NC patients Healthy subjects P
Dendritic cells
SLAM+CD983089983089C+ 983097983094983093 plusmn 983093983094983094 983091983096983088 plusmn 983089983096983096 983088983088983092983093
CD983090983088983093+CD983089983089C+ 983090983089983093983093 plusmn 983089983091983094983092 983092983090983095 plusmn 983091983089983090 983088983088983089983092
IL983091+CD983089983089C+ 983091983093983088983089 plusmn 983090983095983094983089 983091983089983092983096 plusmn 983090983096983095983088 983088983096983089983091
HLA-DR +
CD983089983089C+
983095983089983092983095 plusmn 983089983097983096983095 983094983097983088983093 plusmn 983089983090983097983094 983088983096983088983093CD983096983094+CD983089983089C+ 983091983097983095983088 plusmn 983090983093983096983095 983093983092983090983089 plusmn983089983090983092983093 983088983090983093983088
CD983092983088+CD983089983089C+ 983095983097983090 plusmn 983095983097983096 983090983096983089 plusmn 983089983092983097 983088983089983096983090
cells
CD983092+CD983090983093 High
FoxP983091+CD983089983090983095minuslow 983089983092983090983090 plusmn 983097983088983096 983092983091983093 plusmn 983092983090983094 983088983088983089983097983092
CD983092+ lymphocytes 983091983088983096983093 plusmn 983089983089983092983097 983092983091983093983097 plusmn 983093983094983088 983088983088983095
P-NC patients Among them 1047297gures the SLAMF983089 gene whoseexpressed protein was also ound in dendritic cells rom non-treated NC patients (able 983090) Te other downregulated genes
ound in these patients were MOR NFKB983090 and IL983089983090RB983090(able 983091) In these severe SABIV-NC patients GB983090 andIL983090983092 genes were ound to be up-regulated
4 Discussion
regs play a pivotal role in modulating the host environmentso parasites may 1047297nd more appropriate conditions or theirestablishment and development [983095] It is also possible thatregs may avor the parasite persistence even when a speci1047297ctreatment is used to promote the destruction o the parasite[983090983090] During Taenia solium inection an increase in thelevels o regulatory cells in blood and CSF in NC patients
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 79
Clinical and Developmental Immunology 983095
983137983138983148983141 983091 Genes differentially expressed in NC patients with respectto the parasite location afer 983096 days o treatment
NC caused by cysticerci localized in thesubarachnoid base or intraventricular versus
parenchymal
Genes Log
Downregulated
MOR minus983088983094983097 983088983097983093
NFKB983090 minus983088983097983091 983088983095983096
SLAMF983089 minus983089983088983093 983090983089983096
IL983089983090RB983090 minus983088983096983089 983089983089983097
Upregulated IL983090983092 983088983096983090 983088983097983094
GFB983090 983089983088983094 983090983089983091
Telog old change(LogFC)describeshowmucha quantitychangesromaninitial to a 1047297nal value in a determinate gene B value representsthe possibility that the gene is differentially expressed
has been observed probably promoted by the parasite orcontrolling the central in1047298ammatory environment and thusavoring its survival [983097] Increased regs in NC patients may result rom expanded natural regs or may be induced by theparasite Te latter possibility was explored herein On theother hand DCs may drive cell differentiation to regulatory or effector cells depending on their activation status In thisstudy in vivo and in vitro lines o evidence point to theeffect o cysticercal components on the activation o dendriticcells and their impact on promoting cell differentiation
to CD983090983093high Foxp983091+ CD983092 cells Indeed co-cultivation o dendritic cells with either T solium or T crassiceps cysticerci
promotes a status that avors the differentiation o peripheral cells to CD983092+ regs Comparable results were ound whenperipheral cells rom non-treated NC patients were studied
Te similar effects induced by the presence both o Tsolium or T crassiceps cysticerci merit some comments It iswell known that both cestodes share multiple antigens [983090983091]a act that has been exploited to use the murine cysticercalantigens or diagnosis [983090983092 983090983093]
Parasite-differentiated dendritic cells show no differencewith respect to control in the maturation marker (CD983096983091)neither in the costimulatory molecules HLA-DR CD983096983088CD983096983094 or CD983092983088 these 1047297ndings are compatible with an imma-ture dendritic phenotype In contrast an overexpression o
SLAMF983089 B983095-H983089 and CD983090983088983093 was observed Te latter twomolecules are related to a tolerogenic DC phenotype asreported in many studies [983089983090 983090983094] It is important to notethat the expression o SLAMF983089 B983095-H983089 and CD983090983088983093 wasound accompanied by reg induction in this study as wellHowever their participation in reg induction remains to beelucidatedTis may be particularly relevant since no increasein the two main regulatory cytokines IL983089983088 and GF907317 wasobserved in the supernatants recovered during in vitro reginduction (able 983089) An increased expression o SLAMF983089 and
CD983090983088983093 in dendritic cells and in CD983090983093high Foxp983091+ CD983092 cells was observed in NC patients Both in vitro and invivo 1047297ndings reinorce the relevance o SLAMF983089 and CD983090983088983093
dendritic cells de1047297ning their critical role in cell immunity regulation [983090983095 983090983096]
It is also worth noticing the downexpression o SLAMF983089in the group o 983096 SABIV-NC patients with respect tothose patients harboring cysticerci in the parenchyma Tisapparent discordance with the in vitro results rom non-
treated NC patients can be traced to differences promotedby in vivo or in vitro conditions which may differentially modulate the expression o the SLAM-associated protein(SAP) in lymphocytes Indeed the expression o this adaptorSAP protein promotes the in1047298ammatory response while itsabsence promotes a regulatory environment [983090983096] SLAM-amily receptors presenting cells ligands carry out importantimmunomodulatory unctions by one side they regulatelymphocyte interactions and adhesion [983090983096 983090983097] as well as atolerogenic immune pro1047297le [983089983089 983089983090] Although there is noprevious inormation about the unctions o SLAM receptorsduring NC the results shown in this study point to their pos-sible participation in modulating the in1047298ammatory responsepromoted by cysticidal andor corticosteroid treatmentsChanges ound in MOR IL983089983090RB983090 NFKB983090 and GF-907317 genes match with an immunoregulatory environmentMOR [983091983088] IL983089983090RB983090 [983091983089] and NFKB983090 [983091983090 983091983091] genes codingor proteins that promote an in1047298ammatory environmentare ound downregulated whilst GF-907317 which promotes aregulatory immune response [983095] was up-regulated
5 Conclusions
Overall the results shown in this study reinorce our previous1047297ndings on the relevance o regs in controlling the extento the in1047298ammatory response in NC patients and added in
vivo and in vitro lines o evidence that cysticerci may drivea particular dendritic cell phenotype that induces regulatory cells even though the mechanisms that underlined thisphenomenon remain to be elucidated Additionally whilethe clinical relevance o the promotion o this regulatory environment needs to be evaluated it is easible to proposethat it could avor parasite survival
Acknowledgments
Tis study was supported by CONACY (CB-983090983088983088983096-983088983089983089983088983088983095983088983096 CB-983090983088983089983089-983088983089 983089983094983095983090983095983096 and 983096983094983093983090983095) and by DGAPAIN983090983089983091983097983089983089 Mexico Juan Francisco Rodrıguez prooread the
English version o this paper Te authors thank to MariselaHernandez or her technical assistance and Carlos Castel-lanos Barba or his technical assistance in 1047298ow cytometry
References
[983089] J F Heiber and L Geiger ldquoContext and location depen-dence o adaptive Foxp983091+ regulatory cell ormation duringimmunopathological conditionsrdquo Cellular Immunology vol983090983095983097 no 983089 pp 983094983088ndash983094983093 983090983088983089983090
[983090] I Apostolou A Sarukhan L Klein and H von BoehmerldquoOrigin o regulatory cellswith known speci1047297cityor antigenrdquoNature Immunology vol 983091 no 983096 pp 983095983093983094ndash983095983094983091 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 89
983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 29
983090 Clinical and Developmental Immunology
participate in the control o the in1047298ammatory response sincea negative correlation between the percentage o peripheralregs and activated CD983096+ and CD4+ cells along with adepressed cell prolierative response was observed [983097]However the mechanisms underlying reg induction in NCare still unknown
Dendritic cells play a prominent role in reg inductionin the beginning o the immune response to pathogenseither by promoting the conversion o naıve cells to regsubpopulationsor by expanding the population o preexistingreg cells [983089983088] Mature DCs direct conventional CD983092+ cells tobecome either speci1047297c helper or regulatory cell subsetsdepending on the affinity o their CR to the antigen thestrength o the costimulatory signals provided by antigen-presenting cells (APCs) and the cytokine milieu In additionthe absence o in1047298ammation arrests dendritic cells into animmature or semimature state (iDC) which promotes cell tolerance by conversion o naıve cells into regs [983089983089]iDCs are characterized by expressing MHC II by a highphagocytosis capacity and by low CD983096983088CD983096983094 expressioniDCs also produce IL-983089983088 but neither IL-983089983090 nor NF [983089983088983089983090] Moreover it has been proposed that preexisting regscan educate iDCs to become tolerogenic promoting reggeneration [983089983088 983089983091] Helminths as well as other pathogensmay modulate the immune tolerance properties o DCs [983089983092ndash983089983095]
Tis study was designed to evaluate the capacity o cysticerci to modulate dendritic cells driving them to reginduction both in vitro and in vivo
2 Materials and Methods
983090983089 Parasites Taenia solium metacestodes were obtainedrom naturally inected pigs coming rom villages o Guer-rero an endemic region in Mexico Pigs were euthanizedaccording to ethical animal handling regulations in MexicoCysticerci wereindividually harvested rom muscle tissue andmaintained in RPMI 983089983094983092983088 medium containing 983089983088 etal cal serum and 983089 antibiotics (Invitrogen NY USA) Cysticercio Taenia crassiceps ORF strain were also used in this study to evaluate their potential to induce iDCs and thereoreregs T crassiceps cysticerci were obtained rom inectedBALBcAnN emale mice Parasites were harvested rom theperitoneal cavity o stock emale mice afer 983089983088 weeks o inection
983090983090 Cell Puri1047297cationand DC Generation in thePresenceof LiveCysticerci Human cells were isolated rom buffy coats romblood o healthy donors (Banco de Sangre del Centro MedicoNacional Siglo XXI Mexico DF)
A RosetteSep Human CD983092+ Cell Enrichment Cocktailkit was used or CD983092+ cell puri1047297cation (StemCell Vancou-
ver Canada) Puri1047297ed CD983092+ lymphocytes including gt983097983088o CD983091+ CD983092+ cells were kept rozen at ndash983096983088∘C until used
Monocyte-derived DCs (MDDCs) were isolated rommononuclear cells rom healthy donors using a Rosette-Sep Human Monocyte Enrichment Cocktail kit (StemCellVancouver Canada) MDDCs were generated as previously
reported [983089983096] with only minor modi1047297cations Brie1047298y mono-cytes were plated in Petri dishes (P983089983088983088) at 983093 times 9830899830885 cellsmLin 983089983088 mL o RPMI 983089983094983092983088 medium (Invitrogen NY USA)containing 983089983088 human AB serum (HSAB) and 983089 antibiotics983090983088 ngmL IL-983092 (eBiosciences CA USA) and 983089983088983088 ngmLgranulocyte-macrophage colony-stimulating actor (eBio-
sciences San Diego CA USA) or 983096 days One hal o complete medium including cytokines was replaced at day 983092
o evaluate the role o cysticerci during DC differenti-ation monocytes were cultured as described above eitherwith or without vesicular Taenia crassiceps (983092983088 cysts) orTaenia solium (983093 cysts) cysticerci per dish Te effect o both cestodes on DC differentiation was tested consideringthat murine cysticerci may eventually provide a controlledparasite source o puri1047297ed immune-modulatory componentsDexamethasone at 983089983088minus7 M was used as a positive control
983090983091 In Vitro Treg Induction with Cysticercal Antigens Para-
site-induced MDDCs were seeded at 983090983093 times 9830899830884
cellmL in983089 mL RPMI 983089983094983092983088 medium (Invitrogen NY USA) containing983089983088 human AB serum and 983089 antibiotics (Invitrogen NYUSA) MDDCs were maintained in culture or 983090983092 h Dur-ing this period dexamethasone at 983089983088minus7 M which maintainsa tolerogenic DC phenotype was used as control [983089983097]Parasite-induced DCs were pulsed using 983091 1038389gmL o totalantigens rom T crassiceps cysticerci T crassiceps antigenswere employed considering that T solium and T crassicepscysticerci share in common more than 983097983096 o the expressedantigens [983090983088] 983089983090983093 times 9830899830885 autologous CD983092+ lymphocyteswere cocultured or 983095 days with MDDCs in RPMI 983089983094983092983088medium (Invitrogen NY USA) containing 983089983088 human AB
serum and 983089 antibiotics (Invitrogen NY USA) Tenthe phenotype o regulatory cells was evaluated by 1047298ow cytometry as described below Supernatants o these cultureswere used to evaluate cytokine pro1047297le
983090983092 Cell Phenotype Te ollowing combination o antibodieswasused to characterizethe phenotype o MDDCs(a) MHC-II FIC (mouse IgG983090a k) SLAMF983089 PE (mouse IgG983089 k) CD983089983089cPerCP-eFluor 983095983089983088 (mouse IgG983089 k) and B983095H983089 APC (mouseIgG983089 k) (b) CD983096983091 FIC CD983096983088 PE CD983089983089c PerCP-eFluor 983095983089983088(mouse IgG983089 k) and CD983096983094 APC (mouse IgG983089 k) (c) CD983090983088983093FIC (mouse IgG983090b k) CD983092983088 PE (mouse IgG983089 k) CD983089983089cPerCP-eFluor 983095983089983088 (mouse IgG983089 k) and IL983091 APC (mouse
IgG983089 k) Inducible regulatory cell phenotype was assessedusing CD983089983090983095 FIC (mouse IgG983089 k) Foxp983091 PE (Rat IgG983090a k)CD983092 PerCp (mouse IgG983089 k) and CD983090983093 APC (mouse IgG983089 k)Regulatory cells were intracellularlystained orFoxp983091usingthe eBiosciences kit (most antibodies used or cell stainingwere purchased rom eBiosciences CA USA) All antibodieswere titrated or optimal detection o positive populationspriorto its use considering the manuacturerrsquos recommendedconcentrations
For cytometry analyses cells were 1047297rst gated according tolymphocyte orward and sidelight-scattering propertiesregnumbers were de1047297ned as the raction o Foxp983091+ cells among
CD983092+ CD983090983093high cells
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Clinical and Developmental Immunology 983091
983090983093 Cytokines Cytokine levels in supernatants were mea-sured using the cytometric bead array Cytokine Kit II(BD Biosciences Pharmingen CA USA) according to themanuacturerrsquos instructions in an FACSCalibur cytometerData were analyzed with the BD Cytometric Bead Array sofware (BD Biosciences Pharmingen CA USA) Flow
cytometer was calibrated using BD FACSComp (BD Bio-sciences Pharmingen CA USA) and BD CaliBRIE beads(BD Biosciences Pharmingen CA USA) Assay sensitivitieswere set as ollows IL-983090 (983090983094 pgmL) IL-983094 (983091983088 pgmL) IL-983089983088 (983090983096 pgmL) NF- (983090983096 pgmL) and IFN-1103925 (983095983089 pgmL)GF-907317 were measured using the humanmouse GF-beta983089 ELISA Ready-SE-Go (eBiosciences CA USA) ELISAwas perormed according to the manuacturerrsquos instructionsdetection limit was 983094983088 pgmL All samples were run induplicate
983090983094 DC and Treg Phenotype in NC Patients Blood samplesrom 983089983091 patients who attended at the Instituto Nacional deNeurologıa y Neurocirugıa and Centro Medico NacionalSiglo XXI in Mexico City with con1047297rmed NC diagnosis wereincluded in this study All samples were taken beore any treatment was administered Eight male (age mean 983093983094983090983093 plusmn983091983096 years) and 1047297ve emale (age mean 983091983095 plusmn 983089983094 years) patientswere included in this study
In all cases NC was diagnosed based on clinical manies-tations (seizures ocal de1047297cit and intracranial hypertension)and radiological studies such as MRI and computed tomog-raphy (C)
Vesicular parasites were observed in most cases (983089983090983089983091patients) A solitary cysticercus was seen in 983097 patients andmultiple cysticerci in 983090 patients the rest o the patients
had calci1047297ed and colloidal orms In seven patients par-asite was located at the subarachnoid space o the base(SAB) while the rest o the patients showed parasites atthe cerebral parenchyma (P) Patients with SAB cysticerciexhibited in1047298ammatory traits in cerebral spinal 1047298uid (glucose983091983088983095 plusmn 983090983096 mgdL proteins 983092983090983089 plusmn 983092983088983095 mgdL and 983095983088983095 plusmn
983097983094983097 cellmm3) In contrast patients with parenchymal cys-ticerci showed no in1047298ammation signs Blood samples rom 983093healthy subjects were included as controls
983090983095 Microarrays DNA microarrays were used to comparethe gene expression pro1047297le o mononuclear peripheral cells
(PBMCs) rom a cohort o 983096 NC patients 983092983088 plusmn 983089983092 years oldPBMC rom all patients were obtained 983096 days afer cysticidaltreatment with albendazole 983091983088 mg per kg and concomi-tant corticosteroid administration Tese 983096 (983092 emales and983092 males) patients showed cysticerci in different locationsCysticerci were establishedin the subarachnoid basal space orin the ventricles (SABIV) in 983093 patients and were establishedin the cerebral parenchyma (P) in 983091 patients When requiredor patientsrsquo ollowup CSF was obtained by lumbar punctureat the National Institute o Neurology and Neurosurgery o
Mexico City A mean o 983095983092 cellmm3 (range 983089ndash983090983094983089) was ound
in SABIV-NC patients and a mean o 983094983094 cellmm3 (range983088ndash983089983096) cells was ound in P-NC patients PBMCs were isolated
and then cultured by 983095983090 h with T solium cysticercal antigensobtained as previously described [983090983089]
Recovered cells were placed in RIzol (Gibco NY USA)or RNA puri1047297cation using the RNeasy mini kit (QIAGENX USA) ollowing the manuacturerrsquos instructions Teamount o obtained RNA was estimated in an Agilent 983090983089983088983088
Bioanalyzer (Agilent echnologies Waldbronn Deutsch-land) Comparisons were done with respect to parasitelocation (SABIV-NC versus P-NC) using the R-languageplatorm under the terms o the Free Sofware FoundationsGNU General License (see httpcranr-projectorg) Datawere standardized and normalized All genes were analyzedusing the Reactome sofware (see httpwwwreactomeorg)to 1047297nd a possible biological pathway involved
983090983096 Ethical Considerations Te present study ul1047297lled allregulations or research with human subjects as requiredby the Mexican law and international regulations It alsocomplied with all ethical aspects considered in the General
Rules o Health or Clinical Investigation Ethics Commit-tee at Instituto Nacional de Neurologıa y NeurocirugıaMexico approved the protocol Written inormed consent wasobtained rom all participants Patients were inormed thatsamples obtained would be used or this work
983090983097 Statistical Analysis Data were processed in InStat(GraphPad Sofware Inc CA USA) Variables were describ-ed using mean plusmn SD Differences between groups were cal-culated with Studentsrsquo -test values less than 983088983088983093 wereconsidered signi1047297cant
3 Results
983091983089 Effect of Cysticerci on Dendritic Cell Phenotype Te semi-mature DC phenotype is associated with tolerance and reginduction [983089983090] o evaluate the effect o cysticerci on dendriticcell differentiation the phenotype o DC generated romCD983089983092+ monocytes eitherin thepresence or absence o vesicu-lar cysticerci afer 983096-day culture was studied Dexamethasone(983089983088minus7 M) was employed as a positive control (Figure 983089) Teexpression o CD983096983091+ as well as the costimulatory (CD983096983088CD983096983094 CD983092983088 HLA-DR) and regulatory molecules (SLAMF983089B983095-H983089 CD983090983088983093 and IL983091) on CD983089983089c+ was measured Asshown in Figure 983089 a higher expression o SLAMF983089 B983095-
H983089 and CD983090983088983093 was observed in cells differentiated in thepresence o cysticerci Te expression o HLA-DR CD983096983088CD983096983091 CD983096983094 CD983092983088 and IL983091 molecules did not differbetween control and parasite-driven differentiated DC cellsOn the other hand CD983096983088 CD983092983088 and SLAMF983089 were dimin-ished in dexamethasone-treated cells with respect to control(Figure 983089) HLA-DR and IL983091 expression did not differ romcontrol at any tested condition (data not shown) Altogetherthese results indicate that parasite promotes the expression o molecules related to a DC tolerogenic phenotype
983091983090 Taenia Solium and T crassiceps Cysticerci Promote In VitroRegulatory T Cells Differentiation We investigated whether
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983092 Clinical and Developmental Immunology
Medium Dex
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4053 3155 5606
1351 1393 1839
9944 9477 9951
484 028 129
5380 2739 8256
7455 9987 9933
4733 5947 6413
DC differentiated in the presence of
C D 8 0
C D 8 3
C D 8 6
C D 4 0
S L A M
B 7 - H 1
C D 2 0 5
100
101
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T crassiceps cysticerci
F983145983143983157983154983141 983089 Parasite effect on DC differentiation and activation Monocytes were differentiated to DC in presence o IL-983092 and GM-CSF plusmedium or dexamethasone or Taenia crassicepscysticerci On day 983096 the DC phenotype wasstudiedRepresentative data o three independentexperiments are shown Expression o CD983096983088 CD983096983091 CD983096983094 CD983092983088 SLAMF983089 B983095-H983089 andCD983090983088983093 gated on CD983089983089c+ cells was analyzed by FACSTe isotype controls are shown in white histograms
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Clinical and Developmental Immunology 983093
Pulse
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DCnonstimulated
Dex
P e r c e n t a g e i n c r e a s e o
f r e g u
l a t o r y T c e
l l
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Ag
+
Agminus
T crassiceps T solium
(a)
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C o u n t
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568 448 378 254
208 29 426 344
FOXP3
DC differentiated in the presence of
DC nonstimulated Dex T crassiceps T solium
Pulse
Ag+
Agminus
(b)
F983145983143983157983154983141 983090 In vitro differentiation o dendritic cells in the presence o Taenia solium or T crassiceps cysticerci promotes reg induction (a)Increased percentage in reg cell induction Percentage was calculated as ollows (percentage o regs inducedpercentage o basal regs)times 983089983088983088 Different letters indicate signi1047297cant differences between groups at lt 005 (b) Representative histograms showing Foxp983091 inductionwithin CD983092+ CD983090983093high cells Data are representative o three independent experiments
differentiation in vitro in the presence o live cysticerci stim-
ulated the differentiation to CD983092+ CD983090983093high Foxp983091+ regsHuman DCs either differentiated in the presence or absenceo cysticerci were pulsed or not with cysticercal proteins andused to stimulate autologous CD983092+ lymphocytes Seven
days later the expression o CD983090983093high and Foxp983091 withinCD983092+ lymphocytes was measured As a positive controlo reg induction differentiated DCs were treated withdexamethasone (983089983088minus7 M) Asshown in Figure 983090 cocultivationwith cysticerci signi1047297cantly increased the percentage o CD983092+
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983094 Clinical and Developmental Immunology
983137983138983148983141 983089 Levels o cytokines in supernatants rom in vitro DC differentiation and in vitro regulatory cell induction
IFN-1103925 NF- IL-983089983088 IL-983094 IL-983090 GF-907317
DC nonstimulated 983088983091983097 plusmn 983088983096983096 983091983097983096 plusmn 983091983096983090 983091983089983095 plusmn 983089983095983097 gt983093983088983088983088 983088983091983092 plusmn 983088983095983095 983092983088983094983093983088 plusmn 983089983097983089983094983097daggerControl medium 983089983088 HSAB 983091983093983093 plusmn 983088983093983089 983093983095983096 plusmn 983089983095983097 983093983089983090 plusmn 983089983093983088 gt983093983088983088983088 983089983093983097 plusmn 983089983092983089 NDb
DC differentiation in the presence o
Dexamethasone 983088 983091983089983090 plusmn 983088983092983095 983091983088983096983092 plusmn 983093983096983090a
gt983093983088983088983088 983088 983093983092983093983089983095 plusmn 983090983096983097983091983095Taenia crassiceps 983088983095983096 plusmn 983089983095983093 983090983097983096 plusmn 983089983095983093 983091983095983095 plusmn 983089983095983094 gt983093983088983088983088 983088983095983090 plusmn 983088983097983096 983091983092983089983093983088 plusmn 983090983091983096983089983097
Taenia solium 983088 983092983093983096 plusmn 983092983092983092 983095983091983089 plusmn 983092983096983096 gt983093983088983088983088 983088 983092983091983094983088983088 plusmn 983090983092983091983097983093
reg induction rom naıve CD983092+ cells
DC nonstimulated nonpulsed (Agminus) 983088983095983094 plusmn 983089983091983089 983089983090983089 plusmn 983089983088983095 983088983097983097 plusmn 983088983097983091 983090983093983096 plusmn 983088983095983088 983088983092983093 plusmn 983088983095983096 983089983090983092983094983088 plusmn 983093983091983093
DC nonstimulated pulsed (Ag+) 983088983095983096 plusmn 983089983095983093 983090983088983094 plusmn 983088983096983096 983089983090983090 plusmn 983089983089983090 983090983095983096 plusmn 983088983094983088 983088983092983091 plusmn 983088983097983093 983097983091983088983095 plusmn 983089983094983097983096
Dexamethasone nonpulsed (Agminus) 983088983094983091 plusmn 983089983090983094 983089983094983096 plusmn 983088983091983092 983089983088983089 plusmn 983089983089983096 983090983097983095 plusmn 983090983088983090 983088983091983097 plusmn 983088983095983096 983097983093983092983097 plusmn 983089983097983094983094
Dexamethasone pulsed (Ag+) 983089983088983088 plusmn 983089983095983092 983090983092983097 plusmn 983088983091983094 983090983091983090 plusmn 983089983091983094 983090983095983096 plusmn 983089983089983088 983089983091983096 plusmn 983089983090983095 983097983096983094983091 plusmn 983090983094983091983097
T crassiceps nonpulsed (Agminus) 983089983090983091 plusmn 983090983089983091 983090983088983097 plusmn 983088983095983097 983089983088983090 plusmn 983089983095983094 983096983089983093 plusmn 983089983090983094 983089983091983089 plusmn 983089983090983089 983097983097983090983090 plusmn 983091983089983094983092
T crassiceps pulsed (Ag+) 983089983090983094 plusmn 983089983096983089 983090983089983096 plusmn 983088983091983096 983089983092983092 plusmn 983088983096983089 983090983097983091983092 plusmn 983092983094983095983093 983088983092983091 plusmn 983088983097983093 983089983089983097983096983088 plusmn 983092983090983093983094
T solium nonpulsed (Agminus) 983092983093983088 plusmn 983090983094983089 983090983091983093 plusmn 983088983094983096 983089983092983096 plusmn 983090983089983088 983093983094983097983097 plusmn 983093983090983089 983089983090983089983092 plusmn 983089983089983095983097 983089983089983089983088983088 plusmn 983091983093983092
T solium pulsed (Ag+) 983089983095983091 plusmn 983090983092983093 983090983090983096 plusmn 983088983094983096 983089983097983093 plusmn 983088983088983097 983089983089983094983096 plusmn 983094983088983090 983094983094983089 plusmn 983094983089983094 983095983096983093983088 plusmn 983090983089983090983089
Control cell 983089983093983092 plusmn 983089983092983089 983089983097983088 plusmn 983088983092983096 983089983095983095 plusmn 983088983097983097 983090983088983089 plusmn 983088983092983093 983088983096983096 plusmn 983089983090983088 983089983088983089983088983088 plusmn 983089983094983093983096aSigni1047297cantly different ( lt 005) compared to nonstimulated DC b
Human AB sera rom healthy donors were used in DC differentiation experimentsdaggerLevels o cytokines in the control medium supplemented with 983089983088 HSAB are shown
lymphocytes coexpressing the CD983090983093high and Foxp983091 ractionwith respect to non-stimulated DC rom 983089983089983093 plusmn 983089983091983095 to 983090983089983092983094 plusmn983095983094983091when cocultured with T crassiceps ( = 004)andto983091983090983089983090plusmn 983089983089983093 ( = 005) when cocultured with T solium cysticerciWhen DCs were pulsed with parasite antigens regcells werealso signi1047297cantly increased rom 983089983088983089983090 plusmn 983091983091 to 983090983095983091 plusmn 983096983094983093( = 001) with T crassiceps-differentiated DC cells and to983091983091983097983091 plusmn 983091983096983094 ( = 00002) with T solium-differentiated DCcells (Figure 983090)
983091983091 Cytokine Pro1047297le Te levels o induced cytokines weremeasured in supernatants rom in vitro DC differentiationafer 983096 days o culture and rom in vitro reg inductionafer 983095 days o culture During DC differentiation only IL-983089983088 was signi1047297cantly increased when cells were differentiatedin presence o dexamethasone (able 983089) No difference wasobserved in the other tested conditions
983091983092 Dendritic Cells and Regulatory T Cells in NC PatientsTe in vivo effect o parasite products on the phenotype o peripheral DC rom NC patients was studied Te percentage
o DC andthe expression o regulatory(SLAMF983089 CD983090983088983093 andIL983091) and costimulatory molecules (HLA-DR CD983096983094 andCD983092983088) in CD983089983089c cells in 983089983091 NC patients and 983093 healthy subjectswas measured As able 983090 shows SLAMF983089 and CD983090983088983093 aresigni1047297cantly increased in DC rom NC patients rom 983091983096 to983097983095 and rom 983092983091 to 983090983089983093 respectively ( lt 005) Moreoverperipheral regs are also increased in NC patients rom 983092983091983093to 983089983092983090983090
983091983093 SLAMF983089 Is Downregulated in PBMC from Patients withSABIV Parasites As able 983091 shows only our immune-related genes o the 983091983090983091983090983090 included in the array were ounddownregulated in severe SABIV-NC patients with respect to
983137983138983148983141 983090 Phenotype o peripheral dendritic and regulatory cells inNC patients and healthy subjects
Phenotype NC patients Healthy subjects P
Dendritic cells
SLAM+CD983089983089C+ 983097983094983093 plusmn 983093983094983094 983091983096983088 plusmn 983089983096983096 983088983088983092983093
CD983090983088983093+CD983089983089C+ 983090983089983093983093 plusmn 983089983091983094983092 983092983090983095 plusmn 983091983089983090 983088983088983089983092
IL983091+CD983089983089C+ 983091983093983088983089 plusmn 983090983095983094983089 983091983089983092983096 plusmn 983090983096983095983088 983088983096983089983091
HLA-DR +
CD983089983089C+
983095983089983092983095 plusmn 983089983097983096983095 983094983097983088983093 plusmn 983089983090983097983094 983088983096983088983093CD983096983094+CD983089983089C+ 983091983097983095983088 plusmn 983090983093983096983095 983093983092983090983089 plusmn983089983090983092983093 983088983090983093983088
CD983092983088+CD983089983089C+ 983095983097983090 plusmn 983095983097983096 983090983096983089 plusmn 983089983092983097 983088983089983096983090
cells
CD983092+CD983090983093 High
FoxP983091+CD983089983090983095minuslow 983089983092983090983090 plusmn 983097983088983096 983092983091983093 plusmn 983092983090983094 983088983088983089983097983092
CD983092+ lymphocytes 983091983088983096983093 plusmn 983089983089983092983097 983092983091983093983097 plusmn 983093983094983088 983088983088983095
P-NC patients Among them 1047297gures the SLAMF983089 gene whoseexpressed protein was also ound in dendritic cells rom non-treated NC patients (able 983090) Te other downregulated genes
ound in these patients were MOR NFKB983090 and IL983089983090RB983090(able 983091) In these severe SABIV-NC patients GB983090 andIL983090983092 genes were ound to be up-regulated
4 Discussion
regs play a pivotal role in modulating the host environmentso parasites may 1047297nd more appropriate conditions or theirestablishment and development [983095] It is also possible thatregs may avor the parasite persistence even when a speci1047297ctreatment is used to promote the destruction o the parasite[983090983090] During Taenia solium inection an increase in thelevels o regulatory cells in blood and CSF in NC patients
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Clinical and Developmental Immunology 983095
983137983138983148983141 983091 Genes differentially expressed in NC patients with respectto the parasite location afer 983096 days o treatment
NC caused by cysticerci localized in thesubarachnoid base or intraventricular versus
parenchymal
Genes Log
Downregulated
MOR minus983088983094983097 983088983097983093
NFKB983090 minus983088983097983091 983088983095983096
SLAMF983089 minus983089983088983093 983090983089983096
IL983089983090RB983090 minus983088983096983089 983089983089983097
Upregulated IL983090983092 983088983096983090 983088983097983094
GFB983090 983089983088983094 983090983089983091
Telog old change(LogFC)describeshowmucha quantitychangesromaninitial to a 1047297nal value in a determinate gene B value representsthe possibility that the gene is differentially expressed
has been observed probably promoted by the parasite orcontrolling the central in1047298ammatory environment and thusavoring its survival [983097] Increased regs in NC patients may result rom expanded natural regs or may be induced by theparasite Te latter possibility was explored herein On theother hand DCs may drive cell differentiation to regulatory or effector cells depending on their activation status In thisstudy in vivo and in vitro lines o evidence point to theeffect o cysticercal components on the activation o dendriticcells and their impact on promoting cell differentiation
to CD983090983093high Foxp983091+ CD983092 cells Indeed co-cultivation o dendritic cells with either T solium or T crassiceps cysticerci
promotes a status that avors the differentiation o peripheral cells to CD983092+ regs Comparable results were ound whenperipheral cells rom non-treated NC patients were studied
Te similar effects induced by the presence both o Tsolium or T crassiceps cysticerci merit some comments It iswell known that both cestodes share multiple antigens [983090983091]a act that has been exploited to use the murine cysticercalantigens or diagnosis [983090983092 983090983093]
Parasite-differentiated dendritic cells show no differencewith respect to control in the maturation marker (CD983096983091)neither in the costimulatory molecules HLA-DR CD983096983088CD983096983094 or CD983092983088 these 1047297ndings are compatible with an imma-ture dendritic phenotype In contrast an overexpression o
SLAMF983089 B983095-H983089 and CD983090983088983093 was observed Te latter twomolecules are related to a tolerogenic DC phenotype asreported in many studies [983089983090 983090983094] It is important to notethat the expression o SLAMF983089 B983095-H983089 and CD983090983088983093 wasound accompanied by reg induction in this study as wellHowever their participation in reg induction remains to beelucidatedTis may be particularly relevant since no increasein the two main regulatory cytokines IL983089983088 and GF907317 wasobserved in the supernatants recovered during in vitro reginduction (able 983089) An increased expression o SLAMF983089 and
CD983090983088983093 in dendritic cells and in CD983090983093high Foxp983091+ CD983092 cells was observed in NC patients Both in vitro and invivo 1047297ndings reinorce the relevance o SLAMF983089 and CD983090983088983093
dendritic cells de1047297ning their critical role in cell immunity regulation [983090983095 983090983096]
It is also worth noticing the downexpression o SLAMF983089in the group o 983096 SABIV-NC patients with respect tothose patients harboring cysticerci in the parenchyma Tisapparent discordance with the in vitro results rom non-
treated NC patients can be traced to differences promotedby in vivo or in vitro conditions which may differentially modulate the expression o the SLAM-associated protein(SAP) in lymphocytes Indeed the expression o this adaptorSAP protein promotes the in1047298ammatory response while itsabsence promotes a regulatory environment [983090983096] SLAM-amily receptors presenting cells ligands carry out importantimmunomodulatory unctions by one side they regulatelymphocyte interactions and adhesion [983090983096 983090983097] as well as atolerogenic immune pro1047297le [983089983089 983089983090] Although there is noprevious inormation about the unctions o SLAM receptorsduring NC the results shown in this study point to their pos-sible participation in modulating the in1047298ammatory responsepromoted by cysticidal andor corticosteroid treatmentsChanges ound in MOR IL983089983090RB983090 NFKB983090 and GF-907317 genes match with an immunoregulatory environmentMOR [983091983088] IL983089983090RB983090 [983091983089] and NFKB983090 [983091983090 983091983091] genes codingor proteins that promote an in1047298ammatory environmentare ound downregulated whilst GF-907317 which promotes aregulatory immune response [983095] was up-regulated
5 Conclusions
Overall the results shown in this study reinorce our previous1047297ndings on the relevance o regs in controlling the extento the in1047298ammatory response in NC patients and added in
vivo and in vitro lines o evidence that cysticerci may drivea particular dendritic cell phenotype that induces regulatory cells even though the mechanisms that underlined thisphenomenon remain to be elucidated Additionally whilethe clinical relevance o the promotion o this regulatory environment needs to be evaluated it is easible to proposethat it could avor parasite survival
Acknowledgments
Tis study was supported by CONACY (CB-983090983088983088983096-983088983089983089983088983088983095983088983096 CB-983090983088983089983089-983088983089 983089983094983095983090983095983096 and 983096983094983093983090983095) and by DGAPAIN983090983089983091983097983089983089 Mexico Juan Francisco Rodrıguez prooread the
English version o this paper Te authors thank to MariselaHernandez or her technical assistance and Carlos Castel-lanos Barba or his technical assistance in 1047298ow cytometry
References
[983089] J F Heiber and L Geiger ldquoContext and location depen-dence o adaptive Foxp983091+ regulatory cell ormation duringimmunopathological conditionsrdquo Cellular Immunology vol983090983095983097 no 983089 pp 983094983088ndash983094983093 983090983088983089983090
[983090] I Apostolou A Sarukhan L Klein and H von BoehmerldquoOrigin o regulatory cellswith known speci1047297cityor antigenrdquoNature Immunology vol 983091 no 983096 pp 983095983093983094ndash983095983094983091 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 89
983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 39
Clinical and Developmental Immunology 983091
983090983093 Cytokines Cytokine levels in supernatants were mea-sured using the cytometric bead array Cytokine Kit II(BD Biosciences Pharmingen CA USA) according to themanuacturerrsquos instructions in an FACSCalibur cytometerData were analyzed with the BD Cytometric Bead Array sofware (BD Biosciences Pharmingen CA USA) Flow
cytometer was calibrated using BD FACSComp (BD Bio-sciences Pharmingen CA USA) and BD CaliBRIE beads(BD Biosciences Pharmingen CA USA) Assay sensitivitieswere set as ollows IL-983090 (983090983094 pgmL) IL-983094 (983091983088 pgmL) IL-983089983088 (983090983096 pgmL) NF- (983090983096 pgmL) and IFN-1103925 (983095983089 pgmL)GF-907317 were measured using the humanmouse GF-beta983089 ELISA Ready-SE-Go (eBiosciences CA USA) ELISAwas perormed according to the manuacturerrsquos instructionsdetection limit was 983094983088 pgmL All samples were run induplicate
983090983094 DC and Treg Phenotype in NC Patients Blood samplesrom 983089983091 patients who attended at the Instituto Nacional deNeurologıa y Neurocirugıa and Centro Medico NacionalSiglo XXI in Mexico City with con1047297rmed NC diagnosis wereincluded in this study All samples were taken beore any treatment was administered Eight male (age mean 983093983094983090983093 plusmn983091983096 years) and 1047297ve emale (age mean 983091983095 plusmn 983089983094 years) patientswere included in this study
In all cases NC was diagnosed based on clinical manies-tations (seizures ocal de1047297cit and intracranial hypertension)and radiological studies such as MRI and computed tomog-raphy (C)
Vesicular parasites were observed in most cases (983089983090983089983091patients) A solitary cysticercus was seen in 983097 patients andmultiple cysticerci in 983090 patients the rest o the patients
had calci1047297ed and colloidal orms In seven patients par-asite was located at the subarachnoid space o the base(SAB) while the rest o the patients showed parasites atthe cerebral parenchyma (P) Patients with SAB cysticerciexhibited in1047298ammatory traits in cerebral spinal 1047298uid (glucose983091983088983095 plusmn 983090983096 mgdL proteins 983092983090983089 plusmn 983092983088983095 mgdL and 983095983088983095 plusmn
983097983094983097 cellmm3) In contrast patients with parenchymal cys-ticerci showed no in1047298ammation signs Blood samples rom 983093healthy subjects were included as controls
983090983095 Microarrays DNA microarrays were used to comparethe gene expression pro1047297le o mononuclear peripheral cells
(PBMCs) rom a cohort o 983096 NC patients 983092983088 plusmn 983089983092 years oldPBMC rom all patients were obtained 983096 days afer cysticidaltreatment with albendazole 983091983088 mg per kg and concomi-tant corticosteroid administration Tese 983096 (983092 emales and983092 males) patients showed cysticerci in different locationsCysticerci were establishedin the subarachnoid basal space orin the ventricles (SABIV) in 983093 patients and were establishedin the cerebral parenchyma (P) in 983091 patients When requiredor patientsrsquo ollowup CSF was obtained by lumbar punctureat the National Institute o Neurology and Neurosurgery o
Mexico City A mean o 983095983092 cellmm3 (range 983089ndash983090983094983089) was ound
in SABIV-NC patients and a mean o 983094983094 cellmm3 (range983088ndash983089983096) cells was ound in P-NC patients PBMCs were isolated
and then cultured by 983095983090 h with T solium cysticercal antigensobtained as previously described [983090983089]
Recovered cells were placed in RIzol (Gibco NY USA)or RNA puri1047297cation using the RNeasy mini kit (QIAGENX USA) ollowing the manuacturerrsquos instructions Teamount o obtained RNA was estimated in an Agilent 983090983089983088983088
Bioanalyzer (Agilent echnologies Waldbronn Deutsch-land) Comparisons were done with respect to parasitelocation (SABIV-NC versus P-NC) using the R-languageplatorm under the terms o the Free Sofware FoundationsGNU General License (see httpcranr-projectorg) Datawere standardized and normalized All genes were analyzedusing the Reactome sofware (see httpwwwreactomeorg)to 1047297nd a possible biological pathway involved
983090983096 Ethical Considerations Te present study ul1047297lled allregulations or research with human subjects as requiredby the Mexican law and international regulations It alsocomplied with all ethical aspects considered in the General
Rules o Health or Clinical Investigation Ethics Commit-tee at Instituto Nacional de Neurologıa y NeurocirugıaMexico approved the protocol Written inormed consent wasobtained rom all participants Patients were inormed thatsamples obtained would be used or this work
983090983097 Statistical Analysis Data were processed in InStat(GraphPad Sofware Inc CA USA) Variables were describ-ed using mean plusmn SD Differences between groups were cal-culated with Studentsrsquo -test values less than 983088983088983093 wereconsidered signi1047297cant
3 Results
983091983089 Effect of Cysticerci on Dendritic Cell Phenotype Te semi-mature DC phenotype is associated with tolerance and reginduction [983089983090] o evaluate the effect o cysticerci on dendriticcell differentiation the phenotype o DC generated romCD983089983092+ monocytes eitherin thepresence or absence o vesicu-lar cysticerci afer 983096-day culture was studied Dexamethasone(983089983088minus7 M) was employed as a positive control (Figure 983089) Teexpression o CD983096983091+ as well as the costimulatory (CD983096983088CD983096983094 CD983092983088 HLA-DR) and regulatory molecules (SLAMF983089B983095-H983089 CD983090983088983093 and IL983091) on CD983089983089c+ was measured Asshown in Figure 983089 a higher expression o SLAMF983089 B983095-
H983089 and CD983090983088983093 was observed in cells differentiated in thepresence o cysticerci Te expression o HLA-DR CD983096983088CD983096983091 CD983096983094 CD983092983088 and IL983091 molecules did not differbetween control and parasite-driven differentiated DC cellsOn the other hand CD983096983088 CD983092983088 and SLAMF983089 were dimin-ished in dexamethasone-treated cells with respect to control(Figure 983089) HLA-DR and IL983091 expression did not differ romcontrol at any tested condition (data not shown) Altogetherthese results indicate that parasite promotes the expression o molecules related to a DC tolerogenic phenotype
983091983090 Taenia Solium and T crassiceps Cysticerci Promote In VitroRegulatory T Cells Differentiation We investigated whether
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 49
983092 Clinical and Developmental Immunology
Medium Dex
0
50
99
149
198
0
50
99
149
198
0
50
99
149
198
0
62
124
185
247
0
53
107
160
213
0
45
91
136
181
0
52
105
157
209
0
52
103
155
206
0
59
118
176
235
0
62
124
185
247
0
53
107
160
213
0
45
91
136
181
0
52
105
157
209
0
52
103
155
206
0
59
118
176
235
0
62
124
185
247
0
53
107
160
213
0
45
91
136
181
0
52
105
157
209
0
52
103
155
206
0
59
118
176
235
4053 3155 5606
1351 1393 1839
9944 9477 9951
484 028 129
5380 2739 8256
7455 9987 9933
4733 5947 6413
DC differentiated in the presence of
C D 8 0
C D 8 3
C D 8 6
C D 4 0
S L A M
B 7 - H 1
C D 2 0 5
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
T crassiceps cysticerci
F983145983143983157983154983141 983089 Parasite effect on DC differentiation and activation Monocytes were differentiated to DC in presence o IL-983092 and GM-CSF plusmedium or dexamethasone or Taenia crassicepscysticerci On day 983096 the DC phenotype wasstudiedRepresentative data o three independentexperiments are shown Expression o CD983096983088 CD983096983091 CD983096983094 CD983092983088 SLAMF983089 B983095-H983089 andCD983090983088983093 gated on CD983089983089c+ cells was analyzed by FACSTe isotype controls are shown in white histograms
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 59
Clinical and Developmental Immunology 983093
Pulse
a
bb
bc
ab
bcc
a
0
50
100
150
200
250
300
350
400
450
500
DCnonstimulated
Dex
P e r c e n t a g e i n c r e a s e o
f r e g u
l a t o r y T c e
l l
DC differentiated in the presence of
Ag
+
Agminus
T crassiceps T solium
(a)
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
C o u n t
C o u n t
19
13
6
0
568 448 378 254
208 29 426 344
FOXP3
DC differentiated in the presence of
DC nonstimulated Dex T crassiceps T solium
Pulse
Ag+
Agminus
(b)
F983145983143983157983154983141 983090 In vitro differentiation o dendritic cells in the presence o Taenia solium or T crassiceps cysticerci promotes reg induction (a)Increased percentage in reg cell induction Percentage was calculated as ollows (percentage o regs inducedpercentage o basal regs)times 983089983088983088 Different letters indicate signi1047297cant differences between groups at lt 005 (b) Representative histograms showing Foxp983091 inductionwithin CD983092+ CD983090983093high cells Data are representative o three independent experiments
differentiation in vitro in the presence o live cysticerci stim-
ulated the differentiation to CD983092+ CD983090983093high Foxp983091+ regsHuman DCs either differentiated in the presence or absenceo cysticerci were pulsed or not with cysticercal proteins andused to stimulate autologous CD983092+ lymphocytes Seven
days later the expression o CD983090983093high and Foxp983091 withinCD983092+ lymphocytes was measured As a positive controlo reg induction differentiated DCs were treated withdexamethasone (983089983088minus7 M) Asshown in Figure 983090 cocultivationwith cysticerci signi1047297cantly increased the percentage o CD983092+
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 69
983094 Clinical and Developmental Immunology
983137983138983148983141 983089 Levels o cytokines in supernatants rom in vitro DC differentiation and in vitro regulatory cell induction
IFN-1103925 NF- IL-983089983088 IL-983094 IL-983090 GF-907317
DC nonstimulated 983088983091983097 plusmn 983088983096983096 983091983097983096 plusmn 983091983096983090 983091983089983095 plusmn 983089983095983097 gt983093983088983088983088 983088983091983092 plusmn 983088983095983095 983092983088983094983093983088 plusmn 983089983097983089983094983097daggerControl medium 983089983088 HSAB 983091983093983093 plusmn 983088983093983089 983093983095983096 plusmn 983089983095983097 983093983089983090 plusmn 983089983093983088 gt983093983088983088983088 983089983093983097 plusmn 983089983092983089 NDb
DC differentiation in the presence o
Dexamethasone 983088 983091983089983090 plusmn 983088983092983095 983091983088983096983092 plusmn 983093983096983090a
gt983093983088983088983088 983088 983093983092983093983089983095 plusmn 983090983096983097983091983095Taenia crassiceps 983088983095983096 plusmn 983089983095983093 983090983097983096 plusmn 983089983095983093 983091983095983095 plusmn 983089983095983094 gt983093983088983088983088 983088983095983090 plusmn 983088983097983096 983091983092983089983093983088 plusmn 983090983091983096983089983097
Taenia solium 983088 983092983093983096 plusmn 983092983092983092 983095983091983089 plusmn 983092983096983096 gt983093983088983088983088 983088 983092983091983094983088983088 plusmn 983090983092983091983097983093
reg induction rom naıve CD983092+ cells
DC nonstimulated nonpulsed (Agminus) 983088983095983094 plusmn 983089983091983089 983089983090983089 plusmn 983089983088983095 983088983097983097 plusmn 983088983097983091 983090983093983096 plusmn 983088983095983088 983088983092983093 plusmn 983088983095983096 983089983090983092983094983088 plusmn 983093983091983093
DC nonstimulated pulsed (Ag+) 983088983095983096 plusmn 983089983095983093 983090983088983094 plusmn 983088983096983096 983089983090983090 plusmn 983089983089983090 983090983095983096 plusmn 983088983094983088 983088983092983091 plusmn 983088983097983093 983097983091983088983095 plusmn 983089983094983097983096
Dexamethasone nonpulsed (Agminus) 983088983094983091 plusmn 983089983090983094 983089983094983096 plusmn 983088983091983092 983089983088983089 plusmn 983089983089983096 983090983097983095 plusmn 983090983088983090 983088983091983097 plusmn 983088983095983096 983097983093983092983097 plusmn 983089983097983094983094
Dexamethasone pulsed (Ag+) 983089983088983088 plusmn 983089983095983092 983090983092983097 plusmn 983088983091983094 983090983091983090 plusmn 983089983091983094 983090983095983096 plusmn 983089983089983088 983089983091983096 plusmn 983089983090983095 983097983096983094983091 plusmn 983090983094983091983097
T crassiceps nonpulsed (Agminus) 983089983090983091 plusmn 983090983089983091 983090983088983097 plusmn 983088983095983097 983089983088983090 plusmn 983089983095983094 983096983089983093 plusmn 983089983090983094 983089983091983089 plusmn 983089983090983089 983097983097983090983090 plusmn 983091983089983094983092
T crassiceps pulsed (Ag+) 983089983090983094 plusmn 983089983096983089 983090983089983096 plusmn 983088983091983096 983089983092983092 plusmn 983088983096983089 983090983097983091983092 plusmn 983092983094983095983093 983088983092983091 plusmn 983088983097983093 983089983089983097983096983088 plusmn 983092983090983093983094
T solium nonpulsed (Agminus) 983092983093983088 plusmn 983090983094983089 983090983091983093 plusmn 983088983094983096 983089983092983096 plusmn 983090983089983088 983093983094983097983097 plusmn 983093983090983089 983089983090983089983092 plusmn 983089983089983095983097 983089983089983089983088983088 plusmn 983091983093983092
T solium pulsed (Ag+) 983089983095983091 plusmn 983090983092983093 983090983090983096 plusmn 983088983094983096 983089983097983093 plusmn 983088983088983097 983089983089983094983096 plusmn 983094983088983090 983094983094983089 plusmn 983094983089983094 983095983096983093983088 plusmn 983090983089983090983089
Control cell 983089983093983092 plusmn 983089983092983089 983089983097983088 plusmn 983088983092983096 983089983095983095 plusmn 983088983097983097 983090983088983089 plusmn 983088983092983093 983088983096983096 plusmn 983089983090983088 983089983088983089983088983088 plusmn 983089983094983093983096aSigni1047297cantly different ( lt 005) compared to nonstimulated DC b
Human AB sera rom healthy donors were used in DC differentiation experimentsdaggerLevels o cytokines in the control medium supplemented with 983089983088 HSAB are shown
lymphocytes coexpressing the CD983090983093high and Foxp983091 ractionwith respect to non-stimulated DC rom 983089983089983093 plusmn 983089983091983095 to 983090983089983092983094 plusmn983095983094983091when cocultured with T crassiceps ( = 004)andto983091983090983089983090plusmn 983089983089983093 ( = 005) when cocultured with T solium cysticerciWhen DCs were pulsed with parasite antigens regcells werealso signi1047297cantly increased rom 983089983088983089983090 plusmn 983091983091 to 983090983095983091 plusmn 983096983094983093( = 001) with T crassiceps-differentiated DC cells and to983091983091983097983091 plusmn 983091983096983094 ( = 00002) with T solium-differentiated DCcells (Figure 983090)
983091983091 Cytokine Pro1047297le Te levels o induced cytokines weremeasured in supernatants rom in vitro DC differentiationafer 983096 days o culture and rom in vitro reg inductionafer 983095 days o culture During DC differentiation only IL-983089983088 was signi1047297cantly increased when cells were differentiatedin presence o dexamethasone (able 983089) No difference wasobserved in the other tested conditions
983091983092 Dendritic Cells and Regulatory T Cells in NC PatientsTe in vivo effect o parasite products on the phenotype o peripheral DC rom NC patients was studied Te percentage
o DC andthe expression o regulatory(SLAMF983089 CD983090983088983093 andIL983091) and costimulatory molecules (HLA-DR CD983096983094 andCD983092983088) in CD983089983089c cells in 983089983091 NC patients and 983093 healthy subjectswas measured As able 983090 shows SLAMF983089 and CD983090983088983093 aresigni1047297cantly increased in DC rom NC patients rom 983091983096 to983097983095 and rom 983092983091 to 983090983089983093 respectively ( lt 005) Moreoverperipheral regs are also increased in NC patients rom 983092983091983093to 983089983092983090983090
983091983093 SLAMF983089 Is Downregulated in PBMC from Patients withSABIV Parasites As able 983091 shows only our immune-related genes o the 983091983090983091983090983090 included in the array were ounddownregulated in severe SABIV-NC patients with respect to
983137983138983148983141 983090 Phenotype o peripheral dendritic and regulatory cells inNC patients and healthy subjects
Phenotype NC patients Healthy subjects P
Dendritic cells
SLAM+CD983089983089C+ 983097983094983093 plusmn 983093983094983094 983091983096983088 plusmn 983089983096983096 983088983088983092983093
CD983090983088983093+CD983089983089C+ 983090983089983093983093 plusmn 983089983091983094983092 983092983090983095 plusmn 983091983089983090 983088983088983089983092
IL983091+CD983089983089C+ 983091983093983088983089 plusmn 983090983095983094983089 983091983089983092983096 plusmn 983090983096983095983088 983088983096983089983091
HLA-DR +
CD983089983089C+
983095983089983092983095 plusmn 983089983097983096983095 983094983097983088983093 plusmn 983089983090983097983094 983088983096983088983093CD983096983094+CD983089983089C+ 983091983097983095983088 plusmn 983090983093983096983095 983093983092983090983089 plusmn983089983090983092983093 983088983090983093983088
CD983092983088+CD983089983089C+ 983095983097983090 plusmn 983095983097983096 983090983096983089 plusmn 983089983092983097 983088983089983096983090
cells
CD983092+CD983090983093 High
FoxP983091+CD983089983090983095minuslow 983089983092983090983090 plusmn 983097983088983096 983092983091983093 plusmn 983092983090983094 983088983088983089983097983092
CD983092+ lymphocytes 983091983088983096983093 plusmn 983089983089983092983097 983092983091983093983097 plusmn 983093983094983088 983088983088983095
P-NC patients Among them 1047297gures the SLAMF983089 gene whoseexpressed protein was also ound in dendritic cells rom non-treated NC patients (able 983090) Te other downregulated genes
ound in these patients were MOR NFKB983090 and IL983089983090RB983090(able 983091) In these severe SABIV-NC patients GB983090 andIL983090983092 genes were ound to be up-regulated
4 Discussion
regs play a pivotal role in modulating the host environmentso parasites may 1047297nd more appropriate conditions or theirestablishment and development [983095] It is also possible thatregs may avor the parasite persistence even when a speci1047297ctreatment is used to promote the destruction o the parasite[983090983090] During Taenia solium inection an increase in thelevels o regulatory cells in blood and CSF in NC patients
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 79
Clinical and Developmental Immunology 983095
983137983138983148983141 983091 Genes differentially expressed in NC patients with respectto the parasite location afer 983096 days o treatment
NC caused by cysticerci localized in thesubarachnoid base or intraventricular versus
parenchymal
Genes Log
Downregulated
MOR minus983088983094983097 983088983097983093
NFKB983090 minus983088983097983091 983088983095983096
SLAMF983089 minus983089983088983093 983090983089983096
IL983089983090RB983090 minus983088983096983089 983089983089983097
Upregulated IL983090983092 983088983096983090 983088983097983094
GFB983090 983089983088983094 983090983089983091
Telog old change(LogFC)describeshowmucha quantitychangesromaninitial to a 1047297nal value in a determinate gene B value representsthe possibility that the gene is differentially expressed
has been observed probably promoted by the parasite orcontrolling the central in1047298ammatory environment and thusavoring its survival [983097] Increased regs in NC patients may result rom expanded natural regs or may be induced by theparasite Te latter possibility was explored herein On theother hand DCs may drive cell differentiation to regulatory or effector cells depending on their activation status In thisstudy in vivo and in vitro lines o evidence point to theeffect o cysticercal components on the activation o dendriticcells and their impact on promoting cell differentiation
to CD983090983093high Foxp983091+ CD983092 cells Indeed co-cultivation o dendritic cells with either T solium or T crassiceps cysticerci
promotes a status that avors the differentiation o peripheral cells to CD983092+ regs Comparable results were ound whenperipheral cells rom non-treated NC patients were studied
Te similar effects induced by the presence both o Tsolium or T crassiceps cysticerci merit some comments It iswell known that both cestodes share multiple antigens [983090983091]a act that has been exploited to use the murine cysticercalantigens or diagnosis [983090983092 983090983093]
Parasite-differentiated dendritic cells show no differencewith respect to control in the maturation marker (CD983096983091)neither in the costimulatory molecules HLA-DR CD983096983088CD983096983094 or CD983092983088 these 1047297ndings are compatible with an imma-ture dendritic phenotype In contrast an overexpression o
SLAMF983089 B983095-H983089 and CD983090983088983093 was observed Te latter twomolecules are related to a tolerogenic DC phenotype asreported in many studies [983089983090 983090983094] It is important to notethat the expression o SLAMF983089 B983095-H983089 and CD983090983088983093 wasound accompanied by reg induction in this study as wellHowever their participation in reg induction remains to beelucidatedTis may be particularly relevant since no increasein the two main regulatory cytokines IL983089983088 and GF907317 wasobserved in the supernatants recovered during in vitro reginduction (able 983089) An increased expression o SLAMF983089 and
CD983090983088983093 in dendritic cells and in CD983090983093high Foxp983091+ CD983092 cells was observed in NC patients Both in vitro and invivo 1047297ndings reinorce the relevance o SLAMF983089 and CD983090983088983093
dendritic cells de1047297ning their critical role in cell immunity regulation [983090983095 983090983096]
It is also worth noticing the downexpression o SLAMF983089in the group o 983096 SABIV-NC patients with respect tothose patients harboring cysticerci in the parenchyma Tisapparent discordance with the in vitro results rom non-
treated NC patients can be traced to differences promotedby in vivo or in vitro conditions which may differentially modulate the expression o the SLAM-associated protein(SAP) in lymphocytes Indeed the expression o this adaptorSAP protein promotes the in1047298ammatory response while itsabsence promotes a regulatory environment [983090983096] SLAM-amily receptors presenting cells ligands carry out importantimmunomodulatory unctions by one side they regulatelymphocyte interactions and adhesion [983090983096 983090983097] as well as atolerogenic immune pro1047297le [983089983089 983089983090] Although there is noprevious inormation about the unctions o SLAM receptorsduring NC the results shown in this study point to their pos-sible participation in modulating the in1047298ammatory responsepromoted by cysticidal andor corticosteroid treatmentsChanges ound in MOR IL983089983090RB983090 NFKB983090 and GF-907317 genes match with an immunoregulatory environmentMOR [983091983088] IL983089983090RB983090 [983091983089] and NFKB983090 [983091983090 983091983091] genes codingor proteins that promote an in1047298ammatory environmentare ound downregulated whilst GF-907317 which promotes aregulatory immune response [983095] was up-regulated
5 Conclusions
Overall the results shown in this study reinorce our previous1047297ndings on the relevance o regs in controlling the extento the in1047298ammatory response in NC patients and added in
vivo and in vitro lines o evidence that cysticerci may drivea particular dendritic cell phenotype that induces regulatory cells even though the mechanisms that underlined thisphenomenon remain to be elucidated Additionally whilethe clinical relevance o the promotion o this regulatory environment needs to be evaluated it is easible to proposethat it could avor parasite survival
Acknowledgments
Tis study was supported by CONACY (CB-983090983088983088983096-983088983089983089983088983088983095983088983096 CB-983090983088983089983089-983088983089 983089983094983095983090983095983096 and 983096983094983093983090983095) and by DGAPAIN983090983089983091983097983089983089 Mexico Juan Francisco Rodrıguez prooread the
English version o this paper Te authors thank to MariselaHernandez or her technical assistance and Carlos Castel-lanos Barba or his technical assistance in 1047298ow cytometry
References
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[983090] I Apostolou A Sarukhan L Klein and H von BoehmerldquoOrigin o regulatory cellswith known speci1047297cityor antigenrdquoNature Immunology vol 983091 no 983096 pp 983095983093983094ndash983095983094983091 983090983088983088983090
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983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 49
983092 Clinical and Developmental Immunology
Medium Dex
0
50
99
149
198
0
50
99
149
198
0
50
99
149
198
0
62
124
185
247
0
53
107
160
213
0
45
91
136
181
0
52
105
157
209
0
52
103
155
206
0
59
118
176
235
0
62
124
185
247
0
53
107
160
213
0
45
91
136
181
0
52
105
157
209
0
52
103
155
206
0
59
118
176
235
0
62
124
185
247
0
53
107
160
213
0
45
91
136
181
0
52
105
157
209
0
52
103
155
206
0
59
118
176
235
4053 3155 5606
1351 1393 1839
9944 9477 9951
484 028 129
5380 2739 8256
7455 9987 9933
4733 5947 6413
DC differentiated in the presence of
C D 8 0
C D 8 3
C D 8 6
C D 4 0
S L A M
B 7 - H 1
C D 2 0 5
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
T crassiceps cysticerci
F983145983143983157983154983141 983089 Parasite effect on DC differentiation and activation Monocytes were differentiated to DC in presence o IL-983092 and GM-CSF plusmedium or dexamethasone or Taenia crassicepscysticerci On day 983096 the DC phenotype wasstudiedRepresentative data o three independentexperiments are shown Expression o CD983096983088 CD983096983091 CD983096983094 CD983092983088 SLAMF983089 B983095-H983089 andCD983090983088983093 gated on CD983089983089c+ cells was analyzed by FACSTe isotype controls are shown in white histograms
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 59
Clinical and Developmental Immunology 983093
Pulse
a
bb
bc
ab
bcc
a
0
50
100
150
200
250
300
350
400
450
500
DCnonstimulated
Dex
P e r c e n t a g e i n c r e a s e o
f r e g u
l a t o r y T c e
l l
DC differentiated in the presence of
Ag
+
Agminus
T crassiceps T solium
(a)
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
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100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
C o u n t
C o u n t
19
13
6
0
568 448 378 254
208 29 426 344
FOXP3
DC differentiated in the presence of
DC nonstimulated Dex T crassiceps T solium
Pulse
Ag+
Agminus
(b)
F983145983143983157983154983141 983090 In vitro differentiation o dendritic cells in the presence o Taenia solium or T crassiceps cysticerci promotes reg induction (a)Increased percentage in reg cell induction Percentage was calculated as ollows (percentage o regs inducedpercentage o basal regs)times 983089983088983088 Different letters indicate signi1047297cant differences between groups at lt 005 (b) Representative histograms showing Foxp983091 inductionwithin CD983092+ CD983090983093high cells Data are representative o three independent experiments
differentiation in vitro in the presence o live cysticerci stim-
ulated the differentiation to CD983092+ CD983090983093high Foxp983091+ regsHuman DCs either differentiated in the presence or absenceo cysticerci were pulsed or not with cysticercal proteins andused to stimulate autologous CD983092+ lymphocytes Seven
days later the expression o CD983090983093high and Foxp983091 withinCD983092+ lymphocytes was measured As a positive controlo reg induction differentiated DCs were treated withdexamethasone (983089983088minus7 M) Asshown in Figure 983090 cocultivationwith cysticerci signi1047297cantly increased the percentage o CD983092+
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 69
983094 Clinical and Developmental Immunology
983137983138983148983141 983089 Levels o cytokines in supernatants rom in vitro DC differentiation and in vitro regulatory cell induction
IFN-1103925 NF- IL-983089983088 IL-983094 IL-983090 GF-907317
DC nonstimulated 983088983091983097 plusmn 983088983096983096 983091983097983096 plusmn 983091983096983090 983091983089983095 plusmn 983089983095983097 gt983093983088983088983088 983088983091983092 plusmn 983088983095983095 983092983088983094983093983088 plusmn 983089983097983089983094983097daggerControl medium 983089983088 HSAB 983091983093983093 plusmn 983088983093983089 983093983095983096 plusmn 983089983095983097 983093983089983090 plusmn 983089983093983088 gt983093983088983088983088 983089983093983097 plusmn 983089983092983089 NDb
DC differentiation in the presence o
Dexamethasone 983088 983091983089983090 plusmn 983088983092983095 983091983088983096983092 plusmn 983093983096983090a
gt983093983088983088983088 983088 983093983092983093983089983095 plusmn 983090983096983097983091983095Taenia crassiceps 983088983095983096 plusmn 983089983095983093 983090983097983096 plusmn 983089983095983093 983091983095983095 plusmn 983089983095983094 gt983093983088983088983088 983088983095983090 plusmn 983088983097983096 983091983092983089983093983088 plusmn 983090983091983096983089983097
Taenia solium 983088 983092983093983096 plusmn 983092983092983092 983095983091983089 plusmn 983092983096983096 gt983093983088983088983088 983088 983092983091983094983088983088 plusmn 983090983092983091983097983093
reg induction rom naıve CD983092+ cells
DC nonstimulated nonpulsed (Agminus) 983088983095983094 plusmn 983089983091983089 983089983090983089 plusmn 983089983088983095 983088983097983097 plusmn 983088983097983091 983090983093983096 plusmn 983088983095983088 983088983092983093 plusmn 983088983095983096 983089983090983092983094983088 plusmn 983093983091983093
DC nonstimulated pulsed (Ag+) 983088983095983096 plusmn 983089983095983093 983090983088983094 plusmn 983088983096983096 983089983090983090 plusmn 983089983089983090 983090983095983096 plusmn 983088983094983088 983088983092983091 plusmn 983088983097983093 983097983091983088983095 plusmn 983089983094983097983096
Dexamethasone nonpulsed (Agminus) 983088983094983091 plusmn 983089983090983094 983089983094983096 plusmn 983088983091983092 983089983088983089 plusmn 983089983089983096 983090983097983095 plusmn 983090983088983090 983088983091983097 plusmn 983088983095983096 983097983093983092983097 plusmn 983089983097983094983094
Dexamethasone pulsed (Ag+) 983089983088983088 plusmn 983089983095983092 983090983092983097 plusmn 983088983091983094 983090983091983090 plusmn 983089983091983094 983090983095983096 plusmn 983089983089983088 983089983091983096 plusmn 983089983090983095 983097983096983094983091 plusmn 983090983094983091983097
T crassiceps nonpulsed (Agminus) 983089983090983091 plusmn 983090983089983091 983090983088983097 plusmn 983088983095983097 983089983088983090 plusmn 983089983095983094 983096983089983093 plusmn 983089983090983094 983089983091983089 plusmn 983089983090983089 983097983097983090983090 plusmn 983091983089983094983092
T crassiceps pulsed (Ag+) 983089983090983094 plusmn 983089983096983089 983090983089983096 plusmn 983088983091983096 983089983092983092 plusmn 983088983096983089 983090983097983091983092 plusmn 983092983094983095983093 983088983092983091 plusmn 983088983097983093 983089983089983097983096983088 plusmn 983092983090983093983094
T solium nonpulsed (Agminus) 983092983093983088 plusmn 983090983094983089 983090983091983093 plusmn 983088983094983096 983089983092983096 plusmn 983090983089983088 983093983094983097983097 plusmn 983093983090983089 983089983090983089983092 plusmn 983089983089983095983097 983089983089983089983088983088 plusmn 983091983093983092
T solium pulsed (Ag+) 983089983095983091 plusmn 983090983092983093 983090983090983096 plusmn 983088983094983096 983089983097983093 plusmn 983088983088983097 983089983089983094983096 plusmn 983094983088983090 983094983094983089 plusmn 983094983089983094 983095983096983093983088 plusmn 983090983089983090983089
Control cell 983089983093983092 plusmn 983089983092983089 983089983097983088 plusmn 983088983092983096 983089983095983095 plusmn 983088983097983097 983090983088983089 plusmn 983088983092983093 983088983096983096 plusmn 983089983090983088 983089983088983089983088983088 plusmn 983089983094983093983096aSigni1047297cantly different ( lt 005) compared to nonstimulated DC b
Human AB sera rom healthy donors were used in DC differentiation experimentsdaggerLevels o cytokines in the control medium supplemented with 983089983088 HSAB are shown
lymphocytes coexpressing the CD983090983093high and Foxp983091 ractionwith respect to non-stimulated DC rom 983089983089983093 plusmn 983089983091983095 to 983090983089983092983094 plusmn983095983094983091when cocultured with T crassiceps ( = 004)andto983091983090983089983090plusmn 983089983089983093 ( = 005) when cocultured with T solium cysticerciWhen DCs were pulsed with parasite antigens regcells werealso signi1047297cantly increased rom 983089983088983089983090 plusmn 983091983091 to 983090983095983091 plusmn 983096983094983093( = 001) with T crassiceps-differentiated DC cells and to983091983091983097983091 plusmn 983091983096983094 ( = 00002) with T solium-differentiated DCcells (Figure 983090)
983091983091 Cytokine Pro1047297le Te levels o induced cytokines weremeasured in supernatants rom in vitro DC differentiationafer 983096 days o culture and rom in vitro reg inductionafer 983095 days o culture During DC differentiation only IL-983089983088 was signi1047297cantly increased when cells were differentiatedin presence o dexamethasone (able 983089) No difference wasobserved in the other tested conditions
983091983092 Dendritic Cells and Regulatory T Cells in NC PatientsTe in vivo effect o parasite products on the phenotype o peripheral DC rom NC patients was studied Te percentage
o DC andthe expression o regulatory(SLAMF983089 CD983090983088983093 andIL983091) and costimulatory molecules (HLA-DR CD983096983094 andCD983092983088) in CD983089983089c cells in 983089983091 NC patients and 983093 healthy subjectswas measured As able 983090 shows SLAMF983089 and CD983090983088983093 aresigni1047297cantly increased in DC rom NC patients rom 983091983096 to983097983095 and rom 983092983091 to 983090983089983093 respectively ( lt 005) Moreoverperipheral regs are also increased in NC patients rom 983092983091983093to 983089983092983090983090
983091983093 SLAMF983089 Is Downregulated in PBMC from Patients withSABIV Parasites As able 983091 shows only our immune-related genes o the 983091983090983091983090983090 included in the array were ounddownregulated in severe SABIV-NC patients with respect to
983137983138983148983141 983090 Phenotype o peripheral dendritic and regulatory cells inNC patients and healthy subjects
Phenotype NC patients Healthy subjects P
Dendritic cells
SLAM+CD983089983089C+ 983097983094983093 plusmn 983093983094983094 983091983096983088 plusmn 983089983096983096 983088983088983092983093
CD983090983088983093+CD983089983089C+ 983090983089983093983093 plusmn 983089983091983094983092 983092983090983095 plusmn 983091983089983090 983088983088983089983092
IL983091+CD983089983089C+ 983091983093983088983089 plusmn 983090983095983094983089 983091983089983092983096 plusmn 983090983096983095983088 983088983096983089983091
HLA-DR +
CD983089983089C+
983095983089983092983095 plusmn 983089983097983096983095 983094983097983088983093 plusmn 983089983090983097983094 983088983096983088983093CD983096983094+CD983089983089C+ 983091983097983095983088 plusmn 983090983093983096983095 983093983092983090983089 plusmn983089983090983092983093 983088983090983093983088
CD983092983088+CD983089983089C+ 983095983097983090 plusmn 983095983097983096 983090983096983089 plusmn 983089983092983097 983088983089983096983090
cells
CD983092+CD983090983093 High
FoxP983091+CD983089983090983095minuslow 983089983092983090983090 plusmn 983097983088983096 983092983091983093 plusmn 983092983090983094 983088983088983089983097983092
CD983092+ lymphocytes 983091983088983096983093 plusmn 983089983089983092983097 983092983091983093983097 plusmn 983093983094983088 983088983088983095
P-NC patients Among them 1047297gures the SLAMF983089 gene whoseexpressed protein was also ound in dendritic cells rom non-treated NC patients (able 983090) Te other downregulated genes
ound in these patients were MOR NFKB983090 and IL983089983090RB983090(able 983091) In these severe SABIV-NC patients GB983090 andIL983090983092 genes were ound to be up-regulated
4 Discussion
regs play a pivotal role in modulating the host environmentso parasites may 1047297nd more appropriate conditions or theirestablishment and development [983095] It is also possible thatregs may avor the parasite persistence even when a speci1047297ctreatment is used to promote the destruction o the parasite[983090983090] During Taenia solium inection an increase in thelevels o regulatory cells in blood and CSF in NC patients
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 79
Clinical and Developmental Immunology 983095
983137983138983148983141 983091 Genes differentially expressed in NC patients with respectto the parasite location afer 983096 days o treatment
NC caused by cysticerci localized in thesubarachnoid base or intraventricular versus
parenchymal
Genes Log
Downregulated
MOR minus983088983094983097 983088983097983093
NFKB983090 minus983088983097983091 983088983095983096
SLAMF983089 minus983089983088983093 983090983089983096
IL983089983090RB983090 minus983088983096983089 983089983089983097
Upregulated IL983090983092 983088983096983090 983088983097983094
GFB983090 983089983088983094 983090983089983091
Telog old change(LogFC)describeshowmucha quantitychangesromaninitial to a 1047297nal value in a determinate gene B value representsthe possibility that the gene is differentially expressed
has been observed probably promoted by the parasite orcontrolling the central in1047298ammatory environment and thusavoring its survival [983097] Increased regs in NC patients may result rom expanded natural regs or may be induced by theparasite Te latter possibility was explored herein On theother hand DCs may drive cell differentiation to regulatory or effector cells depending on their activation status In thisstudy in vivo and in vitro lines o evidence point to theeffect o cysticercal components on the activation o dendriticcells and their impact on promoting cell differentiation
to CD983090983093high Foxp983091+ CD983092 cells Indeed co-cultivation o dendritic cells with either T solium or T crassiceps cysticerci
promotes a status that avors the differentiation o peripheral cells to CD983092+ regs Comparable results were ound whenperipheral cells rom non-treated NC patients were studied
Te similar effects induced by the presence both o Tsolium or T crassiceps cysticerci merit some comments It iswell known that both cestodes share multiple antigens [983090983091]a act that has been exploited to use the murine cysticercalantigens or diagnosis [983090983092 983090983093]
Parasite-differentiated dendritic cells show no differencewith respect to control in the maturation marker (CD983096983091)neither in the costimulatory molecules HLA-DR CD983096983088CD983096983094 or CD983092983088 these 1047297ndings are compatible with an imma-ture dendritic phenotype In contrast an overexpression o
SLAMF983089 B983095-H983089 and CD983090983088983093 was observed Te latter twomolecules are related to a tolerogenic DC phenotype asreported in many studies [983089983090 983090983094] It is important to notethat the expression o SLAMF983089 B983095-H983089 and CD983090983088983093 wasound accompanied by reg induction in this study as wellHowever their participation in reg induction remains to beelucidatedTis may be particularly relevant since no increasein the two main regulatory cytokines IL983089983088 and GF907317 wasobserved in the supernatants recovered during in vitro reginduction (able 983089) An increased expression o SLAMF983089 and
CD983090983088983093 in dendritic cells and in CD983090983093high Foxp983091+ CD983092 cells was observed in NC patients Both in vitro and invivo 1047297ndings reinorce the relevance o SLAMF983089 and CD983090983088983093
dendritic cells de1047297ning their critical role in cell immunity regulation [983090983095 983090983096]
It is also worth noticing the downexpression o SLAMF983089in the group o 983096 SABIV-NC patients with respect tothose patients harboring cysticerci in the parenchyma Tisapparent discordance with the in vitro results rom non-
treated NC patients can be traced to differences promotedby in vivo or in vitro conditions which may differentially modulate the expression o the SLAM-associated protein(SAP) in lymphocytes Indeed the expression o this adaptorSAP protein promotes the in1047298ammatory response while itsabsence promotes a regulatory environment [983090983096] SLAM-amily receptors presenting cells ligands carry out importantimmunomodulatory unctions by one side they regulatelymphocyte interactions and adhesion [983090983096 983090983097] as well as atolerogenic immune pro1047297le [983089983089 983089983090] Although there is noprevious inormation about the unctions o SLAM receptorsduring NC the results shown in this study point to their pos-sible participation in modulating the in1047298ammatory responsepromoted by cysticidal andor corticosteroid treatmentsChanges ound in MOR IL983089983090RB983090 NFKB983090 and GF-907317 genes match with an immunoregulatory environmentMOR [983091983088] IL983089983090RB983090 [983091983089] and NFKB983090 [983091983090 983091983091] genes codingor proteins that promote an in1047298ammatory environmentare ound downregulated whilst GF-907317 which promotes aregulatory immune response [983095] was up-regulated
5 Conclusions
Overall the results shown in this study reinorce our previous1047297ndings on the relevance o regs in controlling the extento the in1047298ammatory response in NC patients and added in
vivo and in vitro lines o evidence that cysticerci may drivea particular dendritic cell phenotype that induces regulatory cells even though the mechanisms that underlined thisphenomenon remain to be elucidated Additionally whilethe clinical relevance o the promotion o this regulatory environment needs to be evaluated it is easible to proposethat it could avor parasite survival
Acknowledgments
Tis study was supported by CONACY (CB-983090983088983088983096-983088983089983089983088983088983095983088983096 CB-983090983088983089983089-983088983089 983089983094983095983090983095983096 and 983096983094983093983090983095) and by DGAPAIN983090983089983091983097983089983089 Mexico Juan Francisco Rodrıguez prooread the
English version o this paper Te authors thank to MariselaHernandez or her technical assistance and Carlos Castel-lanos Barba or his technical assistance in 1047298ow cytometry
References
[983089] J F Heiber and L Geiger ldquoContext and location depen-dence o adaptive Foxp983091+ regulatory cell ormation duringimmunopathological conditionsrdquo Cellular Immunology vol983090983095983097 no 983089 pp 983094983088ndash983094983093 983090983088983089983090
[983090] I Apostolou A Sarukhan L Klein and H von BoehmerldquoOrigin o regulatory cellswith known speci1047297cityor antigenrdquoNature Immunology vol 983091 no 983096 pp 983095983093983094ndash983095983094983091 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 89
983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 59
Clinical and Developmental Immunology 983093
Pulse
a
bb
bc
ab
bcc
a
0
50
100
150
200
250
300
350
400
450
500
DCnonstimulated
Dex
P e r c e n t a g e i n c r e a s e o
f r e g u
l a t o r y T c e
l l
DC differentiated in the presence of
Ag
+
Agminus
T crassiceps T solium
(a)
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
100
101
102
103
104
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
19
13
6
0
25
C o u n t
C o u n t
19
13
6
0
568 448 378 254
208 29 426 344
FOXP3
DC differentiated in the presence of
DC nonstimulated Dex T crassiceps T solium
Pulse
Ag+
Agminus
(b)
F983145983143983157983154983141 983090 In vitro differentiation o dendritic cells in the presence o Taenia solium or T crassiceps cysticerci promotes reg induction (a)Increased percentage in reg cell induction Percentage was calculated as ollows (percentage o regs inducedpercentage o basal regs)times 983089983088983088 Different letters indicate signi1047297cant differences between groups at lt 005 (b) Representative histograms showing Foxp983091 inductionwithin CD983092+ CD983090983093high cells Data are representative o three independent experiments
differentiation in vitro in the presence o live cysticerci stim-
ulated the differentiation to CD983092+ CD983090983093high Foxp983091+ regsHuman DCs either differentiated in the presence or absenceo cysticerci were pulsed or not with cysticercal proteins andused to stimulate autologous CD983092+ lymphocytes Seven
days later the expression o CD983090983093high and Foxp983091 withinCD983092+ lymphocytes was measured As a positive controlo reg induction differentiated DCs were treated withdexamethasone (983089983088minus7 M) Asshown in Figure 983090 cocultivationwith cysticerci signi1047297cantly increased the percentage o CD983092+
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 69
983094 Clinical and Developmental Immunology
983137983138983148983141 983089 Levels o cytokines in supernatants rom in vitro DC differentiation and in vitro regulatory cell induction
IFN-1103925 NF- IL-983089983088 IL-983094 IL-983090 GF-907317
DC nonstimulated 983088983091983097 plusmn 983088983096983096 983091983097983096 plusmn 983091983096983090 983091983089983095 plusmn 983089983095983097 gt983093983088983088983088 983088983091983092 plusmn 983088983095983095 983092983088983094983093983088 plusmn 983089983097983089983094983097daggerControl medium 983089983088 HSAB 983091983093983093 plusmn 983088983093983089 983093983095983096 plusmn 983089983095983097 983093983089983090 plusmn 983089983093983088 gt983093983088983088983088 983089983093983097 plusmn 983089983092983089 NDb
DC differentiation in the presence o
Dexamethasone 983088 983091983089983090 plusmn 983088983092983095 983091983088983096983092 plusmn 983093983096983090a
gt983093983088983088983088 983088 983093983092983093983089983095 plusmn 983090983096983097983091983095Taenia crassiceps 983088983095983096 plusmn 983089983095983093 983090983097983096 plusmn 983089983095983093 983091983095983095 plusmn 983089983095983094 gt983093983088983088983088 983088983095983090 plusmn 983088983097983096 983091983092983089983093983088 plusmn 983090983091983096983089983097
Taenia solium 983088 983092983093983096 plusmn 983092983092983092 983095983091983089 plusmn 983092983096983096 gt983093983088983088983088 983088 983092983091983094983088983088 plusmn 983090983092983091983097983093
reg induction rom naıve CD983092+ cells
DC nonstimulated nonpulsed (Agminus) 983088983095983094 plusmn 983089983091983089 983089983090983089 plusmn 983089983088983095 983088983097983097 plusmn 983088983097983091 983090983093983096 plusmn 983088983095983088 983088983092983093 plusmn 983088983095983096 983089983090983092983094983088 plusmn 983093983091983093
DC nonstimulated pulsed (Ag+) 983088983095983096 plusmn 983089983095983093 983090983088983094 plusmn 983088983096983096 983089983090983090 plusmn 983089983089983090 983090983095983096 plusmn 983088983094983088 983088983092983091 plusmn 983088983097983093 983097983091983088983095 plusmn 983089983094983097983096
Dexamethasone nonpulsed (Agminus) 983088983094983091 plusmn 983089983090983094 983089983094983096 plusmn 983088983091983092 983089983088983089 plusmn 983089983089983096 983090983097983095 plusmn 983090983088983090 983088983091983097 plusmn 983088983095983096 983097983093983092983097 plusmn 983089983097983094983094
Dexamethasone pulsed (Ag+) 983089983088983088 plusmn 983089983095983092 983090983092983097 plusmn 983088983091983094 983090983091983090 plusmn 983089983091983094 983090983095983096 plusmn 983089983089983088 983089983091983096 plusmn 983089983090983095 983097983096983094983091 plusmn 983090983094983091983097
T crassiceps nonpulsed (Agminus) 983089983090983091 plusmn 983090983089983091 983090983088983097 plusmn 983088983095983097 983089983088983090 plusmn 983089983095983094 983096983089983093 plusmn 983089983090983094 983089983091983089 plusmn 983089983090983089 983097983097983090983090 plusmn 983091983089983094983092
T crassiceps pulsed (Ag+) 983089983090983094 plusmn 983089983096983089 983090983089983096 plusmn 983088983091983096 983089983092983092 plusmn 983088983096983089 983090983097983091983092 plusmn 983092983094983095983093 983088983092983091 plusmn 983088983097983093 983089983089983097983096983088 plusmn 983092983090983093983094
T solium nonpulsed (Agminus) 983092983093983088 plusmn 983090983094983089 983090983091983093 plusmn 983088983094983096 983089983092983096 plusmn 983090983089983088 983093983094983097983097 plusmn 983093983090983089 983089983090983089983092 plusmn 983089983089983095983097 983089983089983089983088983088 plusmn 983091983093983092
T solium pulsed (Ag+) 983089983095983091 plusmn 983090983092983093 983090983090983096 plusmn 983088983094983096 983089983097983093 plusmn 983088983088983097 983089983089983094983096 plusmn 983094983088983090 983094983094983089 plusmn 983094983089983094 983095983096983093983088 plusmn 983090983089983090983089
Control cell 983089983093983092 plusmn 983089983092983089 983089983097983088 plusmn 983088983092983096 983089983095983095 plusmn 983088983097983097 983090983088983089 plusmn 983088983092983093 983088983096983096 plusmn 983089983090983088 983089983088983089983088983088 plusmn 983089983094983093983096aSigni1047297cantly different ( lt 005) compared to nonstimulated DC b
Human AB sera rom healthy donors were used in DC differentiation experimentsdaggerLevels o cytokines in the control medium supplemented with 983089983088 HSAB are shown
lymphocytes coexpressing the CD983090983093high and Foxp983091 ractionwith respect to non-stimulated DC rom 983089983089983093 plusmn 983089983091983095 to 983090983089983092983094 plusmn983095983094983091when cocultured with T crassiceps ( = 004)andto983091983090983089983090plusmn 983089983089983093 ( = 005) when cocultured with T solium cysticerciWhen DCs were pulsed with parasite antigens regcells werealso signi1047297cantly increased rom 983089983088983089983090 plusmn 983091983091 to 983090983095983091 plusmn 983096983094983093( = 001) with T crassiceps-differentiated DC cells and to983091983091983097983091 plusmn 983091983096983094 ( = 00002) with T solium-differentiated DCcells (Figure 983090)
983091983091 Cytokine Pro1047297le Te levels o induced cytokines weremeasured in supernatants rom in vitro DC differentiationafer 983096 days o culture and rom in vitro reg inductionafer 983095 days o culture During DC differentiation only IL-983089983088 was signi1047297cantly increased when cells were differentiatedin presence o dexamethasone (able 983089) No difference wasobserved in the other tested conditions
983091983092 Dendritic Cells and Regulatory T Cells in NC PatientsTe in vivo effect o parasite products on the phenotype o peripheral DC rom NC patients was studied Te percentage
o DC andthe expression o regulatory(SLAMF983089 CD983090983088983093 andIL983091) and costimulatory molecules (HLA-DR CD983096983094 andCD983092983088) in CD983089983089c cells in 983089983091 NC patients and 983093 healthy subjectswas measured As able 983090 shows SLAMF983089 and CD983090983088983093 aresigni1047297cantly increased in DC rom NC patients rom 983091983096 to983097983095 and rom 983092983091 to 983090983089983093 respectively ( lt 005) Moreoverperipheral regs are also increased in NC patients rom 983092983091983093to 983089983092983090983090
983091983093 SLAMF983089 Is Downregulated in PBMC from Patients withSABIV Parasites As able 983091 shows only our immune-related genes o the 983091983090983091983090983090 included in the array were ounddownregulated in severe SABIV-NC patients with respect to
983137983138983148983141 983090 Phenotype o peripheral dendritic and regulatory cells inNC patients and healthy subjects
Phenotype NC patients Healthy subjects P
Dendritic cells
SLAM+CD983089983089C+ 983097983094983093 plusmn 983093983094983094 983091983096983088 plusmn 983089983096983096 983088983088983092983093
CD983090983088983093+CD983089983089C+ 983090983089983093983093 plusmn 983089983091983094983092 983092983090983095 plusmn 983091983089983090 983088983088983089983092
IL983091+CD983089983089C+ 983091983093983088983089 plusmn 983090983095983094983089 983091983089983092983096 plusmn 983090983096983095983088 983088983096983089983091
HLA-DR +
CD983089983089C+
983095983089983092983095 plusmn 983089983097983096983095 983094983097983088983093 plusmn 983089983090983097983094 983088983096983088983093CD983096983094+CD983089983089C+ 983091983097983095983088 plusmn 983090983093983096983095 983093983092983090983089 plusmn983089983090983092983093 983088983090983093983088
CD983092983088+CD983089983089C+ 983095983097983090 plusmn 983095983097983096 983090983096983089 plusmn 983089983092983097 983088983089983096983090
cells
CD983092+CD983090983093 High
FoxP983091+CD983089983090983095minuslow 983089983092983090983090 plusmn 983097983088983096 983092983091983093 plusmn 983092983090983094 983088983088983089983097983092
CD983092+ lymphocytes 983091983088983096983093 plusmn 983089983089983092983097 983092983091983093983097 plusmn 983093983094983088 983088983088983095
P-NC patients Among them 1047297gures the SLAMF983089 gene whoseexpressed protein was also ound in dendritic cells rom non-treated NC patients (able 983090) Te other downregulated genes
ound in these patients were MOR NFKB983090 and IL983089983090RB983090(able 983091) In these severe SABIV-NC patients GB983090 andIL983090983092 genes were ound to be up-regulated
4 Discussion
regs play a pivotal role in modulating the host environmentso parasites may 1047297nd more appropriate conditions or theirestablishment and development [983095] It is also possible thatregs may avor the parasite persistence even when a speci1047297ctreatment is used to promote the destruction o the parasite[983090983090] During Taenia solium inection an increase in thelevels o regulatory cells in blood and CSF in NC patients
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 79
Clinical and Developmental Immunology 983095
983137983138983148983141 983091 Genes differentially expressed in NC patients with respectto the parasite location afer 983096 days o treatment
NC caused by cysticerci localized in thesubarachnoid base or intraventricular versus
parenchymal
Genes Log
Downregulated
MOR minus983088983094983097 983088983097983093
NFKB983090 minus983088983097983091 983088983095983096
SLAMF983089 minus983089983088983093 983090983089983096
IL983089983090RB983090 minus983088983096983089 983089983089983097
Upregulated IL983090983092 983088983096983090 983088983097983094
GFB983090 983089983088983094 983090983089983091
Telog old change(LogFC)describeshowmucha quantitychangesromaninitial to a 1047297nal value in a determinate gene B value representsthe possibility that the gene is differentially expressed
has been observed probably promoted by the parasite orcontrolling the central in1047298ammatory environment and thusavoring its survival [983097] Increased regs in NC patients may result rom expanded natural regs or may be induced by theparasite Te latter possibility was explored herein On theother hand DCs may drive cell differentiation to regulatory or effector cells depending on their activation status In thisstudy in vivo and in vitro lines o evidence point to theeffect o cysticercal components on the activation o dendriticcells and their impact on promoting cell differentiation
to CD983090983093high Foxp983091+ CD983092 cells Indeed co-cultivation o dendritic cells with either T solium or T crassiceps cysticerci
promotes a status that avors the differentiation o peripheral cells to CD983092+ regs Comparable results were ound whenperipheral cells rom non-treated NC patients were studied
Te similar effects induced by the presence both o Tsolium or T crassiceps cysticerci merit some comments It iswell known that both cestodes share multiple antigens [983090983091]a act that has been exploited to use the murine cysticercalantigens or diagnosis [983090983092 983090983093]
Parasite-differentiated dendritic cells show no differencewith respect to control in the maturation marker (CD983096983091)neither in the costimulatory molecules HLA-DR CD983096983088CD983096983094 or CD983092983088 these 1047297ndings are compatible with an imma-ture dendritic phenotype In contrast an overexpression o
SLAMF983089 B983095-H983089 and CD983090983088983093 was observed Te latter twomolecules are related to a tolerogenic DC phenotype asreported in many studies [983089983090 983090983094] It is important to notethat the expression o SLAMF983089 B983095-H983089 and CD983090983088983093 wasound accompanied by reg induction in this study as wellHowever their participation in reg induction remains to beelucidatedTis may be particularly relevant since no increasein the two main regulatory cytokines IL983089983088 and GF907317 wasobserved in the supernatants recovered during in vitro reginduction (able 983089) An increased expression o SLAMF983089 and
CD983090983088983093 in dendritic cells and in CD983090983093high Foxp983091+ CD983092 cells was observed in NC patients Both in vitro and invivo 1047297ndings reinorce the relevance o SLAMF983089 and CD983090983088983093
dendritic cells de1047297ning their critical role in cell immunity regulation [983090983095 983090983096]
It is also worth noticing the downexpression o SLAMF983089in the group o 983096 SABIV-NC patients with respect tothose patients harboring cysticerci in the parenchyma Tisapparent discordance with the in vitro results rom non-
treated NC patients can be traced to differences promotedby in vivo or in vitro conditions which may differentially modulate the expression o the SLAM-associated protein(SAP) in lymphocytes Indeed the expression o this adaptorSAP protein promotes the in1047298ammatory response while itsabsence promotes a regulatory environment [983090983096] SLAM-amily receptors presenting cells ligands carry out importantimmunomodulatory unctions by one side they regulatelymphocyte interactions and adhesion [983090983096 983090983097] as well as atolerogenic immune pro1047297le [983089983089 983089983090] Although there is noprevious inormation about the unctions o SLAM receptorsduring NC the results shown in this study point to their pos-sible participation in modulating the in1047298ammatory responsepromoted by cysticidal andor corticosteroid treatmentsChanges ound in MOR IL983089983090RB983090 NFKB983090 and GF-907317 genes match with an immunoregulatory environmentMOR [983091983088] IL983089983090RB983090 [983091983089] and NFKB983090 [983091983090 983091983091] genes codingor proteins that promote an in1047298ammatory environmentare ound downregulated whilst GF-907317 which promotes aregulatory immune response [983095] was up-regulated
5 Conclusions
Overall the results shown in this study reinorce our previous1047297ndings on the relevance o regs in controlling the extento the in1047298ammatory response in NC patients and added in
vivo and in vitro lines o evidence that cysticerci may drivea particular dendritic cell phenotype that induces regulatory cells even though the mechanisms that underlined thisphenomenon remain to be elucidated Additionally whilethe clinical relevance o the promotion o this regulatory environment needs to be evaluated it is easible to proposethat it could avor parasite survival
Acknowledgments
Tis study was supported by CONACY (CB-983090983088983088983096-983088983089983089983088983088983095983088983096 CB-983090983088983089983089-983088983089 983089983094983095983090983095983096 and 983096983094983093983090983095) and by DGAPAIN983090983089983091983097983089983089 Mexico Juan Francisco Rodrıguez prooread the
English version o this paper Te authors thank to MariselaHernandez or her technical assistance and Carlos Castel-lanos Barba or his technical assistance in 1047298ow cytometry
References
[983089] J F Heiber and L Geiger ldquoContext and location depen-dence o adaptive Foxp983091+ regulatory cell ormation duringimmunopathological conditionsrdquo Cellular Immunology vol983090983095983097 no 983089 pp 983094983088ndash983094983093 983090983088983089983090
[983090] I Apostolou A Sarukhan L Klein and H von BoehmerldquoOrigin o regulatory cellswith known speci1047297cityor antigenrdquoNature Immunology vol 983091 no 983096 pp 983095983093983094ndash983095983094983091 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 89
983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 69
983094 Clinical and Developmental Immunology
983137983138983148983141 983089 Levels o cytokines in supernatants rom in vitro DC differentiation and in vitro regulatory cell induction
IFN-1103925 NF- IL-983089983088 IL-983094 IL-983090 GF-907317
DC nonstimulated 983088983091983097 plusmn 983088983096983096 983091983097983096 plusmn 983091983096983090 983091983089983095 plusmn 983089983095983097 gt983093983088983088983088 983088983091983092 plusmn 983088983095983095 983092983088983094983093983088 plusmn 983089983097983089983094983097daggerControl medium 983089983088 HSAB 983091983093983093 plusmn 983088983093983089 983093983095983096 plusmn 983089983095983097 983093983089983090 plusmn 983089983093983088 gt983093983088983088983088 983089983093983097 plusmn 983089983092983089 NDb
DC differentiation in the presence o
Dexamethasone 983088 983091983089983090 plusmn 983088983092983095 983091983088983096983092 plusmn 983093983096983090a
gt983093983088983088983088 983088 983093983092983093983089983095 plusmn 983090983096983097983091983095Taenia crassiceps 983088983095983096 plusmn 983089983095983093 983090983097983096 plusmn 983089983095983093 983091983095983095 plusmn 983089983095983094 gt983093983088983088983088 983088983095983090 plusmn 983088983097983096 983091983092983089983093983088 plusmn 983090983091983096983089983097
Taenia solium 983088 983092983093983096 plusmn 983092983092983092 983095983091983089 plusmn 983092983096983096 gt983093983088983088983088 983088 983092983091983094983088983088 plusmn 983090983092983091983097983093
reg induction rom naıve CD983092+ cells
DC nonstimulated nonpulsed (Agminus) 983088983095983094 plusmn 983089983091983089 983089983090983089 plusmn 983089983088983095 983088983097983097 plusmn 983088983097983091 983090983093983096 plusmn 983088983095983088 983088983092983093 plusmn 983088983095983096 983089983090983092983094983088 plusmn 983093983091983093
DC nonstimulated pulsed (Ag+) 983088983095983096 plusmn 983089983095983093 983090983088983094 plusmn 983088983096983096 983089983090983090 plusmn 983089983089983090 983090983095983096 plusmn 983088983094983088 983088983092983091 plusmn 983088983097983093 983097983091983088983095 plusmn 983089983094983097983096
Dexamethasone nonpulsed (Agminus) 983088983094983091 plusmn 983089983090983094 983089983094983096 plusmn 983088983091983092 983089983088983089 plusmn 983089983089983096 983090983097983095 plusmn 983090983088983090 983088983091983097 plusmn 983088983095983096 983097983093983092983097 plusmn 983089983097983094983094
Dexamethasone pulsed (Ag+) 983089983088983088 plusmn 983089983095983092 983090983092983097 plusmn 983088983091983094 983090983091983090 plusmn 983089983091983094 983090983095983096 plusmn 983089983089983088 983089983091983096 plusmn 983089983090983095 983097983096983094983091 plusmn 983090983094983091983097
T crassiceps nonpulsed (Agminus) 983089983090983091 plusmn 983090983089983091 983090983088983097 plusmn 983088983095983097 983089983088983090 plusmn 983089983095983094 983096983089983093 plusmn 983089983090983094 983089983091983089 plusmn 983089983090983089 983097983097983090983090 plusmn 983091983089983094983092
T crassiceps pulsed (Ag+) 983089983090983094 plusmn 983089983096983089 983090983089983096 plusmn 983088983091983096 983089983092983092 plusmn 983088983096983089 983090983097983091983092 plusmn 983092983094983095983093 983088983092983091 plusmn 983088983097983093 983089983089983097983096983088 plusmn 983092983090983093983094
T solium nonpulsed (Agminus) 983092983093983088 plusmn 983090983094983089 983090983091983093 plusmn 983088983094983096 983089983092983096 plusmn 983090983089983088 983093983094983097983097 plusmn 983093983090983089 983089983090983089983092 plusmn 983089983089983095983097 983089983089983089983088983088 plusmn 983091983093983092
T solium pulsed (Ag+) 983089983095983091 plusmn 983090983092983093 983090983090983096 plusmn 983088983094983096 983089983097983093 plusmn 983088983088983097 983089983089983094983096 plusmn 983094983088983090 983094983094983089 plusmn 983094983089983094 983095983096983093983088 plusmn 983090983089983090983089
Control cell 983089983093983092 plusmn 983089983092983089 983089983097983088 plusmn 983088983092983096 983089983095983095 plusmn 983088983097983097 983090983088983089 plusmn 983088983092983093 983088983096983096 plusmn 983089983090983088 983089983088983089983088983088 plusmn 983089983094983093983096aSigni1047297cantly different ( lt 005) compared to nonstimulated DC b
Human AB sera rom healthy donors were used in DC differentiation experimentsdaggerLevels o cytokines in the control medium supplemented with 983089983088 HSAB are shown
lymphocytes coexpressing the CD983090983093high and Foxp983091 ractionwith respect to non-stimulated DC rom 983089983089983093 plusmn 983089983091983095 to 983090983089983092983094 plusmn983095983094983091when cocultured with T crassiceps ( = 004)andto983091983090983089983090plusmn 983089983089983093 ( = 005) when cocultured with T solium cysticerciWhen DCs were pulsed with parasite antigens regcells werealso signi1047297cantly increased rom 983089983088983089983090 plusmn 983091983091 to 983090983095983091 plusmn 983096983094983093( = 001) with T crassiceps-differentiated DC cells and to983091983091983097983091 plusmn 983091983096983094 ( = 00002) with T solium-differentiated DCcells (Figure 983090)
983091983091 Cytokine Pro1047297le Te levels o induced cytokines weremeasured in supernatants rom in vitro DC differentiationafer 983096 days o culture and rom in vitro reg inductionafer 983095 days o culture During DC differentiation only IL-983089983088 was signi1047297cantly increased when cells were differentiatedin presence o dexamethasone (able 983089) No difference wasobserved in the other tested conditions
983091983092 Dendritic Cells and Regulatory T Cells in NC PatientsTe in vivo effect o parasite products on the phenotype o peripheral DC rom NC patients was studied Te percentage
o DC andthe expression o regulatory(SLAMF983089 CD983090983088983093 andIL983091) and costimulatory molecules (HLA-DR CD983096983094 andCD983092983088) in CD983089983089c cells in 983089983091 NC patients and 983093 healthy subjectswas measured As able 983090 shows SLAMF983089 and CD983090983088983093 aresigni1047297cantly increased in DC rom NC patients rom 983091983096 to983097983095 and rom 983092983091 to 983090983089983093 respectively ( lt 005) Moreoverperipheral regs are also increased in NC patients rom 983092983091983093to 983089983092983090983090
983091983093 SLAMF983089 Is Downregulated in PBMC from Patients withSABIV Parasites As able 983091 shows only our immune-related genes o the 983091983090983091983090983090 included in the array were ounddownregulated in severe SABIV-NC patients with respect to
983137983138983148983141 983090 Phenotype o peripheral dendritic and regulatory cells inNC patients and healthy subjects
Phenotype NC patients Healthy subjects P
Dendritic cells
SLAM+CD983089983089C+ 983097983094983093 plusmn 983093983094983094 983091983096983088 plusmn 983089983096983096 983088983088983092983093
CD983090983088983093+CD983089983089C+ 983090983089983093983093 plusmn 983089983091983094983092 983092983090983095 plusmn 983091983089983090 983088983088983089983092
IL983091+CD983089983089C+ 983091983093983088983089 plusmn 983090983095983094983089 983091983089983092983096 plusmn 983090983096983095983088 983088983096983089983091
HLA-DR +
CD983089983089C+
983095983089983092983095 plusmn 983089983097983096983095 983094983097983088983093 plusmn 983089983090983097983094 983088983096983088983093CD983096983094+CD983089983089C+ 983091983097983095983088 plusmn 983090983093983096983095 983093983092983090983089 plusmn983089983090983092983093 983088983090983093983088
CD983092983088+CD983089983089C+ 983095983097983090 plusmn 983095983097983096 983090983096983089 plusmn 983089983092983097 983088983089983096983090
cells
CD983092+CD983090983093 High
FoxP983091+CD983089983090983095minuslow 983089983092983090983090 plusmn 983097983088983096 983092983091983093 plusmn 983092983090983094 983088983088983089983097983092
CD983092+ lymphocytes 983091983088983096983093 plusmn 983089983089983092983097 983092983091983093983097 plusmn 983093983094983088 983088983088983095
P-NC patients Among them 1047297gures the SLAMF983089 gene whoseexpressed protein was also ound in dendritic cells rom non-treated NC patients (able 983090) Te other downregulated genes
ound in these patients were MOR NFKB983090 and IL983089983090RB983090(able 983091) In these severe SABIV-NC patients GB983090 andIL983090983092 genes were ound to be up-regulated
4 Discussion
regs play a pivotal role in modulating the host environmentso parasites may 1047297nd more appropriate conditions or theirestablishment and development [983095] It is also possible thatregs may avor the parasite persistence even when a speci1047297ctreatment is used to promote the destruction o the parasite[983090983090] During Taenia solium inection an increase in thelevels o regulatory cells in blood and CSF in NC patients
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 79
Clinical and Developmental Immunology 983095
983137983138983148983141 983091 Genes differentially expressed in NC patients with respectto the parasite location afer 983096 days o treatment
NC caused by cysticerci localized in thesubarachnoid base or intraventricular versus
parenchymal
Genes Log
Downregulated
MOR minus983088983094983097 983088983097983093
NFKB983090 minus983088983097983091 983088983095983096
SLAMF983089 minus983089983088983093 983090983089983096
IL983089983090RB983090 minus983088983096983089 983089983089983097
Upregulated IL983090983092 983088983096983090 983088983097983094
GFB983090 983089983088983094 983090983089983091
Telog old change(LogFC)describeshowmucha quantitychangesromaninitial to a 1047297nal value in a determinate gene B value representsthe possibility that the gene is differentially expressed
has been observed probably promoted by the parasite orcontrolling the central in1047298ammatory environment and thusavoring its survival [983097] Increased regs in NC patients may result rom expanded natural regs or may be induced by theparasite Te latter possibility was explored herein On theother hand DCs may drive cell differentiation to regulatory or effector cells depending on their activation status In thisstudy in vivo and in vitro lines o evidence point to theeffect o cysticercal components on the activation o dendriticcells and their impact on promoting cell differentiation
to CD983090983093high Foxp983091+ CD983092 cells Indeed co-cultivation o dendritic cells with either T solium or T crassiceps cysticerci
promotes a status that avors the differentiation o peripheral cells to CD983092+ regs Comparable results were ound whenperipheral cells rom non-treated NC patients were studied
Te similar effects induced by the presence both o Tsolium or T crassiceps cysticerci merit some comments It iswell known that both cestodes share multiple antigens [983090983091]a act that has been exploited to use the murine cysticercalantigens or diagnosis [983090983092 983090983093]
Parasite-differentiated dendritic cells show no differencewith respect to control in the maturation marker (CD983096983091)neither in the costimulatory molecules HLA-DR CD983096983088CD983096983094 or CD983092983088 these 1047297ndings are compatible with an imma-ture dendritic phenotype In contrast an overexpression o
SLAMF983089 B983095-H983089 and CD983090983088983093 was observed Te latter twomolecules are related to a tolerogenic DC phenotype asreported in many studies [983089983090 983090983094] It is important to notethat the expression o SLAMF983089 B983095-H983089 and CD983090983088983093 wasound accompanied by reg induction in this study as wellHowever their participation in reg induction remains to beelucidatedTis may be particularly relevant since no increasein the two main regulatory cytokines IL983089983088 and GF907317 wasobserved in the supernatants recovered during in vitro reginduction (able 983089) An increased expression o SLAMF983089 and
CD983090983088983093 in dendritic cells and in CD983090983093high Foxp983091+ CD983092 cells was observed in NC patients Both in vitro and invivo 1047297ndings reinorce the relevance o SLAMF983089 and CD983090983088983093
dendritic cells de1047297ning their critical role in cell immunity regulation [983090983095 983090983096]
It is also worth noticing the downexpression o SLAMF983089in the group o 983096 SABIV-NC patients with respect tothose patients harboring cysticerci in the parenchyma Tisapparent discordance with the in vitro results rom non-
treated NC patients can be traced to differences promotedby in vivo or in vitro conditions which may differentially modulate the expression o the SLAM-associated protein(SAP) in lymphocytes Indeed the expression o this adaptorSAP protein promotes the in1047298ammatory response while itsabsence promotes a regulatory environment [983090983096] SLAM-amily receptors presenting cells ligands carry out importantimmunomodulatory unctions by one side they regulatelymphocyte interactions and adhesion [983090983096 983090983097] as well as atolerogenic immune pro1047297le [983089983089 983089983090] Although there is noprevious inormation about the unctions o SLAM receptorsduring NC the results shown in this study point to their pos-sible participation in modulating the in1047298ammatory responsepromoted by cysticidal andor corticosteroid treatmentsChanges ound in MOR IL983089983090RB983090 NFKB983090 and GF-907317 genes match with an immunoregulatory environmentMOR [983091983088] IL983089983090RB983090 [983091983089] and NFKB983090 [983091983090 983091983091] genes codingor proteins that promote an in1047298ammatory environmentare ound downregulated whilst GF-907317 which promotes aregulatory immune response [983095] was up-regulated
5 Conclusions
Overall the results shown in this study reinorce our previous1047297ndings on the relevance o regs in controlling the extento the in1047298ammatory response in NC patients and added in
vivo and in vitro lines o evidence that cysticerci may drivea particular dendritic cell phenotype that induces regulatory cells even though the mechanisms that underlined thisphenomenon remain to be elucidated Additionally whilethe clinical relevance o the promotion o this regulatory environment needs to be evaluated it is easible to proposethat it could avor parasite survival
Acknowledgments
Tis study was supported by CONACY (CB-983090983088983088983096-983088983089983089983088983088983095983088983096 CB-983090983088983089983089-983088983089 983089983094983095983090983095983096 and 983096983094983093983090983095) and by DGAPAIN983090983089983091983097983089983089 Mexico Juan Francisco Rodrıguez prooread the
English version o this paper Te authors thank to MariselaHernandez or her technical assistance and Carlos Castel-lanos Barba or his technical assistance in 1047298ow cytometry
References
[983089] J F Heiber and L Geiger ldquoContext and location depen-dence o adaptive Foxp983091+ regulatory cell ormation duringimmunopathological conditionsrdquo Cellular Immunology vol983090983095983097 no 983089 pp 983094983088ndash983094983093 983090983088983089983090
[983090] I Apostolou A Sarukhan L Klein and H von BoehmerldquoOrigin o regulatory cellswith known speci1047297cityor antigenrdquoNature Immunology vol 983091 no 983096 pp 983095983093983094ndash983095983094983091 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 89
983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 79
Clinical and Developmental Immunology 983095
983137983138983148983141 983091 Genes differentially expressed in NC patients with respectto the parasite location afer 983096 days o treatment
NC caused by cysticerci localized in thesubarachnoid base or intraventricular versus
parenchymal
Genes Log
Downregulated
MOR minus983088983094983097 983088983097983093
NFKB983090 minus983088983097983091 983088983095983096
SLAMF983089 minus983089983088983093 983090983089983096
IL983089983090RB983090 minus983088983096983089 983089983089983097
Upregulated IL983090983092 983088983096983090 983088983097983094
GFB983090 983089983088983094 983090983089983091
Telog old change(LogFC)describeshowmucha quantitychangesromaninitial to a 1047297nal value in a determinate gene B value representsthe possibility that the gene is differentially expressed
has been observed probably promoted by the parasite orcontrolling the central in1047298ammatory environment and thusavoring its survival [983097] Increased regs in NC patients may result rom expanded natural regs or may be induced by theparasite Te latter possibility was explored herein On theother hand DCs may drive cell differentiation to regulatory or effector cells depending on their activation status In thisstudy in vivo and in vitro lines o evidence point to theeffect o cysticercal components on the activation o dendriticcells and their impact on promoting cell differentiation
to CD983090983093high Foxp983091+ CD983092 cells Indeed co-cultivation o dendritic cells with either T solium or T crassiceps cysticerci
promotes a status that avors the differentiation o peripheral cells to CD983092+ regs Comparable results were ound whenperipheral cells rom non-treated NC patients were studied
Te similar effects induced by the presence both o Tsolium or T crassiceps cysticerci merit some comments It iswell known that both cestodes share multiple antigens [983090983091]a act that has been exploited to use the murine cysticercalantigens or diagnosis [983090983092 983090983093]
Parasite-differentiated dendritic cells show no differencewith respect to control in the maturation marker (CD983096983091)neither in the costimulatory molecules HLA-DR CD983096983088CD983096983094 or CD983092983088 these 1047297ndings are compatible with an imma-ture dendritic phenotype In contrast an overexpression o
SLAMF983089 B983095-H983089 and CD983090983088983093 was observed Te latter twomolecules are related to a tolerogenic DC phenotype asreported in many studies [983089983090 983090983094] It is important to notethat the expression o SLAMF983089 B983095-H983089 and CD983090983088983093 wasound accompanied by reg induction in this study as wellHowever their participation in reg induction remains to beelucidatedTis may be particularly relevant since no increasein the two main regulatory cytokines IL983089983088 and GF907317 wasobserved in the supernatants recovered during in vitro reginduction (able 983089) An increased expression o SLAMF983089 and
CD983090983088983093 in dendritic cells and in CD983090983093high Foxp983091+ CD983092 cells was observed in NC patients Both in vitro and invivo 1047297ndings reinorce the relevance o SLAMF983089 and CD983090983088983093
dendritic cells de1047297ning their critical role in cell immunity regulation [983090983095 983090983096]
It is also worth noticing the downexpression o SLAMF983089in the group o 983096 SABIV-NC patients with respect tothose patients harboring cysticerci in the parenchyma Tisapparent discordance with the in vitro results rom non-
treated NC patients can be traced to differences promotedby in vivo or in vitro conditions which may differentially modulate the expression o the SLAM-associated protein(SAP) in lymphocytes Indeed the expression o this adaptorSAP protein promotes the in1047298ammatory response while itsabsence promotes a regulatory environment [983090983096] SLAM-amily receptors presenting cells ligands carry out importantimmunomodulatory unctions by one side they regulatelymphocyte interactions and adhesion [983090983096 983090983097] as well as atolerogenic immune pro1047297le [983089983089 983089983090] Although there is noprevious inormation about the unctions o SLAM receptorsduring NC the results shown in this study point to their pos-sible participation in modulating the in1047298ammatory responsepromoted by cysticidal andor corticosteroid treatmentsChanges ound in MOR IL983089983090RB983090 NFKB983090 and GF-907317 genes match with an immunoregulatory environmentMOR [983091983088] IL983089983090RB983090 [983091983089] and NFKB983090 [983091983090 983091983091] genes codingor proteins that promote an in1047298ammatory environmentare ound downregulated whilst GF-907317 which promotes aregulatory immune response [983095] was up-regulated
5 Conclusions
Overall the results shown in this study reinorce our previous1047297ndings on the relevance o regs in controlling the extento the in1047298ammatory response in NC patients and added in
vivo and in vitro lines o evidence that cysticerci may drivea particular dendritic cell phenotype that induces regulatory cells even though the mechanisms that underlined thisphenomenon remain to be elucidated Additionally whilethe clinical relevance o the promotion o this regulatory environment needs to be evaluated it is easible to proposethat it could avor parasite survival
Acknowledgments
Tis study was supported by CONACY (CB-983090983088983088983096-983088983089983089983088983088983095983088983096 CB-983090983088983089983089-983088983089 983089983094983095983090983095983096 and 983096983094983093983090983095) and by DGAPAIN983090983089983091983097983089983089 Mexico Juan Francisco Rodrıguez prooread the
English version o this paper Te authors thank to MariselaHernandez or her technical assistance and Carlos Castel-lanos Barba or his technical assistance in 1047298ow cytometry
References
[983089] J F Heiber and L Geiger ldquoContext and location depen-dence o adaptive Foxp983091+ regulatory cell ormation duringimmunopathological conditionsrdquo Cellular Immunology vol983090983095983097 no 983089 pp 983094983088ndash983094983093 983090983088983089983090
[983090] I Apostolou A Sarukhan L Klein and H von BoehmerldquoOrigin o regulatory cellswith known speci1047297cityor antigenrdquoNature Immunology vol 983091 no 983096 pp 983095983093983094ndash983095983094983091 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 89
983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 89
983096 Clinical and Developmental Immunology
[983091] A N Akbar L S aams M Salmon and M Vukmanovic-Stejic ldquoTe peripheral generation o CD983092+ CD983090983093+ regulatory cellsrdquo Immunology vol 983089983088983097 no 983091 pp 983091983089983097ndash983091983090983093 983090983088983088983091
[983092] L W Collison V Chaturvedi A L Henderson et al ldquoIL-983091983093-mediated induction o a potent regulatory cell populationrdquoNature Immunology vol 983089983089 no 983089983090 pp 983089983088983097983091ndash983089983089983088983089 983090983088983089983088
[983093] R M Maizels A Balic N Gomez-Escobar M Nair MD aylor and J E Allen ldquoHelminth parasitesmdashmasters o regulationrdquo Immunological Reviews vol 983090983088983089 pp 983096983097ndash983089983089983094 983090983088983088983092
[983094] R M Maizels and M Yazdanbakhsh ldquoImmune regulation by helminth parasites cellular and molecular mechanismsrdquo NatureReviews Immunology vol 983091 no 983097 pp 983095983091983091ndash983095983092983092 983090983088983088983091
[983095] L Adalid-Peralta G Fragoso A Fleury and E Sciutto ldquoMech-anisms underlying the induction o regulatory cells andits relevance in the adaptive immune response in parasiticinectionsrdquo International Journal of Biological Sciences vol 983095 no983097 pp 983089983092983089983090ndash983089983092983090983094 983090983088983089983089
[983096] R M Maizels E J Pearce DArtis M Yazdanbakhsh and AWynn ldquoRegulation o pathogenesis and immunity in helminthinectionsrdquo Journal of Experimental Medicine vol 983090983088983094 no 983089983088pp 983090983088983093983097ndash983090983088983094983094 983090983088983088983097
[983097] L Adalid-Peralta A Fleury M Garcia-Ibarra et al ldquoHumanneurocysticercosis in vivo expansion o peripheral regulatory cells and their recruitment in the central nervous systemrdquo Journal of Parasitology vol 983097983096 no 983089 pp 983089983092983090ndash983089983092983096 983090983088983089983090
[983089983088] R A Maldonado and U H von Andrian ldquoHow tolerogenicdendritic cells induce regulatory cellsrdquo Advances in Immunol-ogy vol 983089983088983096 no C pp 983089983089983089ndash983089983094983093 983090983088983089983088
[983089983089] H H Smits E C De Jong E A Wierenga and M LKapsenberg ldquoDifferent aces o regulatory DCs in homeostasisand immunityrdquo Trends in Immunology vol 983090983094 no 983091 pp 983089983090983091ndash983089983090983097 983090983088983088983093
[983089983090] S Rutella S Danese andG Leoneldquoolerogenic dendriticcellscytokine modulation comes o agerdquo Blood vol 983089983088983096 no 983093 pp
983089983092983091983093ndash983089983092983092983088 983090983088983088983094[983089983091] M Kornete and C A Piccirillo ldquoFunctional crosstalk between
dendritic cellsand Foxp983091+ regulatory cellsin the maintenanceo immune tolerancerdquo Frontiers in Immunology vol 983091 article983089983094983093 983090983088983089983090
[983089983092] P Mukherjee and V S Chauhan ldquoPlasmodium alciparum-reemerozoites and inected RBCs distinctly affect soluble CD983092983088ligand-mediated maturation o immature monocyte-deriveddendritic cellsrdquo Journal of Leukocyte Biology vol 983096983092 no 983089 pp983090983092983092ndash983090983093983092 983090983088983088983096
[983089983093] M Segura Z Su C Piccirillo and M M Stevenson ldquoImpair-ment o dendritic cell unction by excretory-secretory productsa potential mechanism or nematode-induced immunosuppres-sionrdquo European Journal of Immunology vol 983091983095 no 983095 pp 983089983096983096983095ndash
983089983097983088983092 983090983088983088983095[983089983094] D Van der Kleij E Latz J F H M Brouwers et al
ldquoA novel host-parasite lipid cross-talk Schistosomal lyso-phosphatidylserine activates toll-like receptor 983090 and affectsimmune polarizationrdquo Journal of Biological Chemistry vol 983090983095983095no 983093983088 pp 983092983096983089983090983090ndash983092983096983089983090983097 983090983088983088983090
[983089983095] C C Keller O Yamo C Ouma et al ldquoAcquisition o hemo-zoin by monocytes down-regulates interleukin-983089983090 p983092983088 (IL-983089983090p983092983088) transcripts and circulating IL-983089983090p983095983088 through an IL-983089983088-dependent mechanism in vivo and in vitro 1047297ndings in severemalarial anemiardquo Infection and Immunity vol 983095983092 no 983097 pp983093983090983092983097ndash983093983090983094983088 983090983088983088983094
[983089983096] N Cohen E Mouly H Hamdi et al ldquoGILZ expression inhuman dendritic cells redirects their maturation and prevents
antigen-speci1047297c lymphocyte responserdquo Blood vol 983089983088983095 no 983093pp 983090983088983091983095ndash983090983088983092983092 983090983088983088983094
[983089983097] G B Ferreira F S Kleijwegt E Waelkens et al ldquoDifferentialprotein pathwaysin 983089 983090983093-dihydroxyvitamin D3 and dexametha-sone modulated tolerogenic human dendritic cellsrdquo Journal of Proteome Research vol 983089983089 no 983090 pp 983097983092983089ndash983097983095983089 983090983088983089983090
[983090983088] C Larralde R M Montoya E Sciutto M L Diaz Govezen-sky and E Coltorti ldquoDeciphering western blots o tapewormantigens (Taenia solium Echinococcus granulosus and Taeniacrassiceps) reacting with sera rom neurocysticercosis andhydatid disease patientsrdquo American Journal of Tropical Medicineand Hygiene vol 983092983088 no 983091 pp 983090983096983090ndash983090983097983088 983089983097983096983097
[983090983089] A Chavarria A Fleury R J Bobes J Morales G Fragoso andE Sciutto ldquoA depressed peripheral cellular immune responseis related to symptomatic neurocysticercosisrdquo Microbes and Infection vol 983096 no 983092 pp 983089983088983096983090ndash983089983088983096983097 983090983088983088983094
[983090983090] A Verma K N Prasad S S Cheekatla K K Nyati V KPaliwal and R K Gupta ldquoImmune response in symptomaticand asymptomatic neurocysticercosisrdquo Medical Microbiology and Immunology vol 983090983088983088 no 983092 pp 983090983093983093ndash983090983094983089 983090983088983089983089
[983090983091] E Sciutto G Fragoso L rueba et al ldquoCysticercosis vac-cine cross protecting immunity with T solium antigensagainst experimental murine T crassiceps cysticercosisrdquo Para-site Immunology vol 983089983090 no 983094 pp 983094983096983095ndash983094983097983094 983089983097983097983088
[983090983092] G C Arruda A D Da Silva E M A B Quagliato MA Maretti and C L Rossi ldquoEvaluation o Taenia solium andTaenia crassiceps cysticercal antigens or the serodiagnosis o neurocysticercosisrdquo Tropical Medicine and International Health vol 983089983088 no 983089983088 pp 983089983088983088983093ndash983089983088983089983090 983090983088983088983093
[983090983093] R H S Peralta A J Vaz A Pardini et al ldquoEvaluation o anantigen rom Taenia crassiceps cysticercus or the serodiagnosiso neurocysticercosisrdquo Acta Tropica vol 983096983091 no 983090 pp 983089983093983097ndash983089983094983096983090983088983088983090
[983090983094] H Hamdi V Godot M C Maillot et al ldquoInduction o antigen-
speci1047297c regulatory lymphocytes by human dendritic cellsexpressing the glucocorticoid-induced leucine zipperrdquo Blood vol 983089983089983088 no 983089 pp 983090983089983089ndash983090983089983097 983090983088983088983095
[983090983095] Fukaya R Murakami H akagi et al ldquoConditional ablationo CD983090983088983093+ conventional dendritic cells impacts the regulationo -cell immunity and homeostasis in vivordquo Proceedings of theNational Academy of Sciences of the United States of America vol 983089983088983097 no 983090983096 pp 983089983089983090983096983096ndash983089983089983090983097983091 983090983088983089983090
[983090983096] P L Schwartzberg K L Mueller H Qi and J L CannonsldquoSLAM receptors and SAP in1047298uence lymphocyte interactionsdevelopment and unctionrdquo Nature Reviews Immunology vol983097 no 983089 pp 983091983097ndash983092983094 983090983088983088983097
[983090983097] A Veillette Z Dong and S Latour ldquoConsequence o the
SLAM-SAP signaling pathway in innate-like and conventionallymphocytesrdquo Immunity vol 983090983095 no 983093 pp 983094983097983096ndash983095983089983088 983090983088983088983095
[983091983088] J H Lee J P Lydon and C H Kim ldquoProgesterone suppressesthe mOR pathway and promotes generation o induced reg-ulatory cells with increased stabilityrdquo European Journal of Immunology vol 983092983090 no 983089983088 pp 983090983094983096983091ndash983090983094983097983094 983090983088983089983090
[983091983089] F Sinigaglia D DrsquoAmbrosio P Panina-Bordignon and LRogge ldquoRegulation o the IL-983089983090IL-983089983090R axis a critical step in -helper cell differentiation and effector unctionrdquo Immunological Reviews vol 983089983095983088 pp 983094983093ndash983095983090 983089983097983097983097
[983091983090] E F Lind C L Ahonen A Wasiuk et al ldquoDendritic cellsrequire the NF-B983090 pathway or cross-presentation o solubleantigensrdquo Journal of Immunology vol 983089983096983089 no 983089 pp 983091983093983092ndash983091983094983091983090983088983088983096
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
8122019 Inmunidad en La Neurocisticercosis
httpslidepdfcomreaderfullinmunidad-en-la-neurocisticercosis 99
Clinical and Developmental Immunology 983097
[983091983091] D Artis S Shapira N Mason et al ldquoDifferential requirementor NF-B amily members in control o helminth inectionandintestinal in1047298ammationrdquo Journal of Immunology vol 983089983094983097 no 983096pp 983092983092983096983089ndash983092983092983096983095 983090983088983088983090
