RESUMO
A major objective of current HIV-1 vaccination strategies is the induction of HIV-1-specific CD8+ MHC class I-restricted CTL responses, which are suggested to play a pivotal role in viral clearance and protection against HIV-1 disease progression. However, the marked genetic diversity of HIV-1 and existence of distinct viral subtypes or clades could potentially hinder the development of a universally efficacious HIV-1 vaccine. In this study we examined HIV-1 intraclade (B(LAI) versus B(MN)) Env gp160-specific CTL reactivity in recently HIV-1 clade B-infected individuals. We further evaluated the extent of interclade CTL cross-recognition of the divergent A and C Env gp160 subtypes, that are highly prevalent in the global pandemic. Freshly isolated PBMCs were stimulated in vitro with autologous PBMCs infected with recombinant vaccinia vectors expressing HIV-1 env, gag, pol, and nef genes derived from HIV-1 clade B. All 13 of the 19 HIV-1-seropositive subjects who elicited significant clade B Env gp160LAI CD8+ CTL responses also demonstrated comparable levels of CTL cross-reactivity against clade C92BR025 Env gp160. Nine of these individuals also showed extensive interclade CTL cross-recognition of clade A92UG037 Env gp160. Two HLA class I B7 donors had nondetectable intraclade CTL response against B Env gp160MN, while generating significant intraclade B(LAI) and interclade (A and C) Env gp160 CTL cross-reactivity. These observations serve to underscore the central importance of the HLA background of individuals in determining the pattern of immune reactivity to natural HIV-1 infection and presumably vaccines. Five donors studied also demonstrated broad CTL cross-reactivity against clade A92UG037 Gag p55, Pol, and/or Nef antigens. In conclusion, this present study indicates that there is a considerable degree of CD8+ CTL cross-recognition of the highly divergent HIV-1 Env gp160 subtypes during early phases of HIV-1 infection. Such findings suggest that HIV-1 vaccines based on a single clade that can induce extensive cross-clade immunity may demonstrate utility in diverse geographical regions.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Proteína gp160 do Envelope de HIV/imunologia , Infecções por HIV/imunologia , HIV-1 , Linfócitos T Citotóxicos/imunologia , Clonagem Molecular , Reações Cruzadas , Feminino , Produtos do Gene gag/genética , Produtos do Gene gag/imunologia , Produtos do Gene nef/genética , Produtos do Gene nef/imunologia , Produtos do Gene pol/genética , Produtos do Gene pol/imunologia , Antígenos HIV/imunologia , Soropositividade para HIV , Teste de Histocompatibilidade , Humanos , Masculino , Precursores de Proteínas/imunologia , Vaccinia virus/genética , Carga Viral , Produtos do Gene nef do Vírus da Imunodeficiência HumanaRESUMO
Purified human Beta-2 microglobulin (beta 2m) and a human beta 2m fluorochrome conjugate were used in exchange reactions to demonstrate that beta 2m associates with a teleostean cell surface protein. beta 2m exchange among brown bullhead, channel catfish, fathead minnow, and rainbow trout cells lines was detected by using either radioimmunoassay or flow cytometry. Evidence that beta 2m binds specifically with the surface of teleostean cells and possibly associates with an expressed class I MHC homologue is provided. Moreover, following exchange on brown bullhead cells, a coprecipitated protein of 45 kDa was observed following subsequent immunoprecipitation with the human beta 2m specific antibody B1.1G6. Given that beta 2m is a peripheral protein which has been shown to exchange with MHC expressing cells from different species, co-precipitation results suggest that the 45 kDa protein may represent a class I MHC homologue.
