inflammation in IRIS. Experiments are in progress attemptingto delineate a possible role for CD73 dysfunction in theimmuno-pathology of IRIS.

doi:10.1016/j.clim.2008.03.359

Su.9. Effects of Different Forms of FHA on IL-10 andIL-12 Secretion by Human Dendritic CellsViolette Dirix,1 Virginie Verscheure,1 Nathalie Mielcarek,2

Anne-Sophie Debrie,2 Camille Locht,2 Françoise Mascart. 11ULB, Brussels, Belgium; 2Institut Pasteur de Lille, Lille,France

Bordetella pertussis filamentous haemagglutinin (FHA)is a protective antigen included in all pertussis vaccines. B.pertussis infection is associated with an interferon-gamma(IFN-γ) secretion by the circulating lymphocytes in responseto B.pertussis antigens, including FHA, and this cytokinesecretion may be one of the “correlates of protection”against B.pertussis. We have shown that this IFN-γ responseappears only in approximately half of B.pertussis-vaccinatedinfants having received the first three vaccine doses. Thepoor IFN-γ response in the other half of the infants wasrelated to ex-vivo secretion of IL-10 by their monocytes.Since FHA has been shown to induce IL-10 secretion by mousedendritic cells (DCs), we investigated the ability of differentforms of FHA to induce IL-10 secretion by human DCs.Incubation of human DCs with FHA also resulted in thesecretion of significant amounts of IL-10 in more than half ofthe subjects tested. A modified form of FHA, in which oneamino acid has been replaced within the RGD sequenceresponsible for the interaction of FHA with monocytes, alsoinduced IL-10 secretion by DCs. In contrast, a truncatedform, consisting of the 80-kDa N-terminal moiety of FHA,previously shown to be protective in mouse challengemodels, did not induce IL-10 in any of the DCs preparationstested, although it induced IL-12p70. These different formsof FHA will now be tested in vitro for their ability to induceIFN-γ secretion by human lymphocytes from immunizedsubjects in mixed lymphocyte cultures comprising FHA-loaded DCs and allogenic lymphocytes.

doi:10.1016/j.clim.2008.03.360

Su.10. Single Nucleotide Polymorphism onthe -383TNFR1 in Patients with HCV Associated withthe Development of Diabetes Mellitus Type 2Jorge Segura Ortega,1 Laura Moreno-Luna,2 MargaritaMontoya-Buelna,1 Cecilia Guillen-Vargas,1 MaryFafutis-Morris. 1 1Universidad de Guadalajara,Guadalajara, Mexico; 2Antiguo Hospital Civil deGuadalajara, Guadalajara, Mexico

Hepatitis C virus infection (HCV) is a major public healthproblem. HCV infected patients develop DM2 in a highpercentage. The mechanisms remain to be unknown. TNFαhas been identified as an insulin resistance mediator andthis is induced by HCV. We examine the TNFαR1 in 4

different groups: patients with HCV, patients with DM2,patients with HCV and DM and a control group. The aim ofthis study was to detect the -383 TNFαR1 polymorphism.We used the PCR-RFLPs technique. We measure the solubleTNFαR1 levels with ELISA assay. The statistic analysis wasperformed with and SPSS v. 10.0 and “t “Student test.Results: The population was found in Hardy Weinberg'sequilibrium. The frequency of the -383 TNFR1 genotypes C/C, A/C and A/A in DM2 patients was: 0,100, 0%, in HCVpatients was 4.76, 90.48 y 4.76%, in patients DM2+VHC 0.100 y 0% and in the control group was: 0, 47.06 y 52.94%respectively. In the three groups DM2, HCV and DM2 + HCVpatients presented increased serum levels in the solubleTNFR1 the difference was p=0.001 when the three groupswere compared to the control group. Conclusion: The A/Cgenotype was the most frequent of the three study groups.The A/A genotype was the most frequent at the controlgroup with a 52.94% frequency, compared to 4.76% of theHCV patients and in any of the patients of the other groups.This genotype could have a relationship with a decreasedrisk to develop DM2 in Mexican mestizo people.

doi:10.1016/j.clim.2008.03.361

Su.11. Selective Blocking of Toll Like Receptor-9Enhances Expression of Toll Like Receptor-3Pathway Factors During Innate Immune Response inMonocytes Exposed to Human CytomegalovirusChristopher Harrison, Hooi Yew. Children's Mercy Hospital,Kansas City, MO

We previously described the expression of intracellularsignaling factors for Toll Like Receptor (TLR)-2, TLR-3, andTLR-9 pathways after exposure of THP-1 monocyte cells tohuman cytomegalovirus (HCMV). These studies indicatedpotential redundancy of functional response. We usedmonoclonal antibody (Moab) to block TLR-3 and TLR-9 inorder to delineate contributions of each to innate immuneresponses to HCMV. We exposed 500,000 THP-1 monocyticcells to moderately virulent Toledo strain HCMV at a multi-plicity of infection of 10:1, and used unexposed THP-1 cellsas controls. Extracted RNA was assayed by real time PCR forrelative mRNA expression of key factors in the TLR-2 path-way (IL-12), the TLR-3 pathway (TRIF, IRF-3, and type 1interferon(IFNb)) and TLR-9 pathway (NF-kB and TNF-a).Incubation of HCMV exposed THP-1 cells with Moab to TLR-3: 1) Markedly reduced the expected enhancement of mRNAexpression for TLR-3, TRIF, IRF-3 and IFNb at 1 hour. 2) Hadno detectable effect on TLR-2 or TLR-9 pathway factors.Incubation of HCMV exposed THP-1 cells with TLR-9 Moab:1) Yielded a comparable reduction in the expectedenhancement of mRNA expression for TLR9, NF-kB, andTNFa at 1 hour. 2) Had no effect on TLR-2 pathway factors'mRNA. 3) However it exaggerated the expected enhance-ment of mRNA expression for TLR-3 pathway factors.Cytokine release into supernatants appeared to followmRNA expression. These data suggest a compensatoryresponse in the TLR-3 pathway when the TLR-9 pathway isblocked. This interplay of TLR-9 and TLR-3 may be simplecompensation by an independent pathway of endosomal

S127Abstracts

origin or it may indicate that TLR-3 has an upstreamfunction in facilitating TLR-9 access/response to the CPGDNA of HCMV.

doi:10.1016/j.clim.2008.03.362

Su.12. Effect of Short Term Antiretroviral DrugIntervention on CD4+ Cells and Immunoglobulinsin HIV Sero-positive SubjectsMartin Ifeanyichukwu. Nnamdi Azikiwe University, NnewiCampus, Nigeria

Aim: The Aimstudy was designed to assess the effect ofshort-term antiretroviral therapy (ART) on IgA, IgG and IgMand CD4+T cell count in HIV seropositive subjects. Method:20 confirmed HIV seropositive subjects, aged between 15-65 years were recruited for the study. They were on triplecombinations therapy consisting zidovudine, lamivudineand nevirapine. 20 HIV seronegative subjects were used ascontrol. Blood sample was collected from the participantsfor the determination of the above parameters in bothgroups. Result: The CD4+T cell counts show no significantdifference between pre-ART and 2 months post-ARD(PN0.05) but was significantly higher by 4 months post-ART compared with the pre-ART (pb0.05). IgG and IgMserum levels showed significantly high values by 2 and4 months post- ART compared with the pre-ART value(pb0.05 in each case). However, the serum IgA level by4 months post-ART showed no significant differencecompared with pre-ARD value (pN0.05). Meanwhile therewere no significant differences in CD4 count, IgM, IgA, andIgG levels between 4 months post-ART values comparedwith the corresponding values in HIV seronegative controlsubjects. Discussion and conclusion: The present studyshowed an improvement in the blood concentration ofCD4 cell by 4 months post-ART administration, whichsuggests possible recovery of cellular immunity. Theinsignificant difference in IgA concentration within thestudy period possibly suggests non progressive mucosal orsub mucosal infections. Similarly the raised IgM and IgGconcentrations within the study period may be anindication of existing infections and signifies possiblepotentials towards short term recovery. This shows thatwith the use of these drugs prognosis seem good for theshort term.

doi:10.1016/j.clim.2008.03.363

Su.13. NCF1 Gene and Pseudogene Pattern:Association with Parasitic Infectionand AutoimmunityBernhard Greve,1 Peter Hoffmann,1 Reinhard Vonthein,2

Jürgen Kun,2 Bertrand Lell,2 Marcin Mycko,3 Krysztof Selmaj,3

Klaus Berger,4 Robert Weissert,1 Peter Kremsner.2 1HertieInstitute for Clinical Brain Research, Tübingen, Germany;2University of Tübingen, Tübingen, Germany; 3University ofLodz, Lodz, Poland; 4University of Münster, Münster, Germany

Neutrophil cytosolic factor 1, p47phox (NCF1) is acomponent of the NADPH oxidase complex mediatingformation of reactive oxygen radicals (ROI) which play animportant role in host defense and autoimmunity. Anindividual genomic pattern of NCF1 and its two types ofpseudogenes (reflected by the ΔGT/GTGT ratio) mayinfluence the individual capacity to produce ROI. Weexamined the association of ΔGT/GTGT ratios withclinical parameters and individual ROI production in 152Gabonese children with Plasmodium falciparum malaria.We found ΔGT/GTGT ratios weakly associated with ROIproduction from whole blood in children with severemalaria. Multiple sclerosis (MS) patients (n=461) fromGermany and Poland and control persons (n=375) did notdiffer in their ΔGT/GTGT ratios, but we detected anassociation with the age-of-onset among patients withMS. We conclude that the genomic pattern of NCF1 andits two types of pseudogenes might influence ROIproduction but only marginally influence susceptibilityto and outcome of malaria and MS.

doi:10.1016/j.clim.2008.03.364

Su.14. Immunostimulatory Activity ofFullerene DerivativesAlexander Babakhin,1 Alla Garmanova,1 Anna Petrukhina,1

Valentina Romanova,2 Sergey Andreev,1 LawrenceDuBuske.3 1Institute of Immunology, Moscow, RussianFederation; 2Nesmeyanov Institute of OrganoelementCompounds, Moscow, Russian Federation; 3ImmunologyResearch Institute of New England, Gardner, MA

Introduction: The immune system does not recognize ful-lerene C60 spheroids as foreign structures. However, in someexperiments the water-soluble C60-aminocaproic acid (C60-Acp) mixed with ovalbumin (OVA) augmented anti-OVA IgGresponses. This study assesses the potential immunostimu-latory effects of fullerenes covalently bound to peptides andproteins as antigens. Methods: Proteins including bovineserum albumen (BSA), Keyhole Limpet Hemocyanine (KLH)and OVA, or peptides, including fragments of protein E7 fromHuman Papilloma Virus types 16 and 31 which mimic B celland T cell epitopes, were conjugated to water-soluble C60-Acp or C60-maleimide/L-prolinate Na. The immunizationstudies were performed as intra-peritoneal injections intomice using three successive injections. Sera were assayed byELISA to assess for IgG anti-fullerene carrier and anti-antigenantibodies. Results: The IgG assays demonstrated no specificanti-C60 IgG production. Augmentation of anti-peptide oranti-protein antibody production was not observed, theimmunogenicity of unmodified and C60-modified antigensbeing essentially identical. Conclusions: Fullerene com-pounds do not stimulate or suppress IgG-related immuneresponses. The excellent membrane penetration andabsence of immune toxicity make fullerene compoundspotential candidates as novel carriers for drug or vaccinedelivery.

doi:10.1016/j.clim.2008.03.365

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