title: Complement-mediated enhancement of HIV-1 neutralisation by anti-HLA antibodies derived from polytransfused patients.
abstract: We have recently shown that 'alloimmune sera' derived from polytransfused patients (PTP sera) are able to recognise and neutralise HIV in vitro. In this study we try to identify the protein(s), which are recognised by the PTP sera and elucidate mechanisms responsible for the neutralising capacity of these sera. The PTP sera allowed immunoprecipitation (IP) of HLA class II molecules on HIV-infected cells. To detect a potential cross-reactivity of alloreactive antibodies (Ab) with the HIV envelope protein gp160 or its subunits gp120/gp41 and HLA proteins, ELISA and FACS analyses were performed. The lack of reactivity of the PTP sera against rsgp160 in ELISA or FACS analysis indicated that recognition of cells was independent of HIV infection. To clarify whether interaction of the PTP sera with target cells has any effect on the infection process, virus neutralisation assays were performed. Inhibition of HIV infection was observed only when virus was pre-incubated with the PTP sera. Complement enhanced neutralisation of HIV-1 significantly. This enhancement was not due to complement-mediated lysis, because pre-incubation of the target cells with PTP sera did not inhibit HIV replication. Therefore, the neutralising effect of the Ab was due to blocking of the viral attachment/fusion process and not to negative signalling after infection. Since steric hindrance is possible only when HLA and gp120/gp41 are in close vicinity, isolation of rafts and IP assays were performed. These experiments revealed that gp120 and MHC class II molecules are indeed co-localised. The close physical association of gp120/gp41 and HLA strongly supports a mechanism for neutralisation of HIV by anti-HLA-Ab based on steric hindrance.
authors.affiliation: Institute of Hygiene and Social Medicine, Innsbruck, Austria.
authors.forename: Doris
authors.initials: D
authors.lastname: Wilfingseder
authors.forename: Martin
authors.initials: M
authors.lastname: Spruth
authors.forename: Christoph G
authors.initials: CG
authors.lastname: Ammann
authors.forename: Susanne
authors.initials: S
authors.lastname: Döpper
authors.forename: Cornelia
authors.initials: C
authors.lastname: Speth
authors.forename: Manfred P
authors.initials: MP
authors.lastname: Dierich
authors.forename: Heribert
authors.initials: H
authors.lastname: Stoiber
mesh_terms: D000937:Antigen-Antibody Reactions / Q000502:physiology; D015703:Antigens, CD / Q000276:immunology; D003167:Complement Activation / Q000276:immunology; D003165:Complement System Proteins / Q000276:immunology*; D003429:Cross Reactions / Q000276:immunology; D015483:HIV Antibodies / Q000097:blood / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D019302:HIV Envelope Protein gp160 / Q000276:immunology; D015658:HIV Infections / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006680:HLA Antigens / Q000097:blood / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007518:Isoantibodies / Q000276:immunology; D009500:Neutralization Tests; D011233:Precipitin Tests
chemical_list: D015703:Antigens, CD; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D006680:HLA Antigens; D000949:Histocompatibility Antigens Class II; D007074:Immunoglobulin G; D007518:Isoantibodies; D003165:Complement System Proteins