Generation and Characterization of a Defective HIV-1 Virus as an Immunogen for a Therapeutic Vaccine 英文参考文献.docVIP

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Generation and Characterization of a Defective HIV-1 Virus as an Immunogen for a Therapeutic Vaccine 英文参考文献.doc

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Generation and Characterization of a Defective HIV-1 Virus as an Immunogen for a Therapeutic Vaccine 英文参考文献

GenerationandCharacterizationofaDefectiveHIV-1 VirusasanImmunogenforaTherapeuticVaccine CarmenAlvarez-Ferna′ ′ndez1.,AlbertoCrespoGuardo1.,JavierGarc?′a-Pe′rez2,FelipeGarc?′a1, JuliaBlanco3,LauraEscriba`-Garc?′a1,JoseMariaGatell1,JoseAlcam?′2,MontserratPlana1, SonsolesSa′nchez-Palomino1* 1InstitutdInvestigationsBiome`diquesAugustPiiSunyer(IDIBAPS)-AIDSResearchGroup,HospitalClinic,CatalonianCenterforHIVVaccines(HIVACAT)andUniversityof Barcelona, Barcelona, Spain, 2AIDS Immunopathology Unit. National Center of Microbiology, Instituto de Salud Carlos III, Madrid, Spain, 3Institut de Recerca de la SindromedeInmunodeficenciaAdquirida,IrsiCaixa,Badalona,Spain-HIVACAT Abstract Background: The generation of new immunogens able to elicit strong specific immune responses remains a major challengeintheattemptstoobtainaprophylacticortherapeuticvaccineagainstHIV/AIDS.Wedesignedandconstructed a defective recombinant virus based on the HIV-1 genome generating infective but non-replicative virions able to elicit broadandstrongcellularimmuneresponsesinHIV-1seropositiveindividuals. Results:Viralparticlesweregeneratedthroughtransienttransfectioninproducercells(293-T)ofafulllengthHIV-1DNA carrying a deletion of 892 base pairs (bp) in the pol gene encompassing the sequence that codes for the reverse transcriptase (NL4-3/DRT clone). The viral particles generated were able to enter target cells, but due to the absence of reversetranscriptasenoreplicationwasdetected.TheimmunogeniccapacityoftheseparticleswasassessedbyELISPOTto determine c-interferon production in a cohort of 69 chronic asymptomatic HIV-1 seropositive individuals. Surprisingly, defectiveparticlesproducedfromNL4-3/DRTtriggeredstrongercellularresponsesthanwild-typeHIV-1virusesinactivated withAldrithiol-2(AT-2)andinalargerproportionofindividuals(55%versus23%seropositiveindividualstested).Electron microscopy showed that NL4-3/DRT virions display immature morphology. Interestingly, wild-type viruses treated with Ampren