This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.To generate strong cytotoxic T lymphocytes (CTL) responses, in the absence of other immune responses, using several different vaccine approaches.There are still 3 vaccinees remaining from the previous experiment. Two of them have continued to maintain viral loads to constant levels for 3+ years. One animal previously had very low viral loads, but over the past year the viral load has gradually crept up and is now at 30,000 virus particles/ml plasma. We continue to monitor these animals.We started the next experiment in this series, vaccination with all proteins except Env. The previous vaccination experiment utilized Gag, Tat, Rev and Nef. We have now added Pol, Vif, Vpr and Vpx. In April 2007, 8 Mamu-A*02+ animals were vaccinated four times at four week intervals with DNA encoding all eight proteins. In October, 2007, the same animals were boosted with Ad5 encoding all eight proteins. We observed very robust responses to vaccination. While each animal did not make responses to all proteins, overall all proteins were responded to and each animal made responses to a subset of proteins. About half of the responses could be attributed to epitopes that can bind to Mamu-A*02, the remainder are presented by other MHC alleles. We are currently using viral suppression assays to assess the efficacy of the generated responses. In April 2008, we will begin low dose challenges of these animals.This research uses WNPRC Immunology and Virology Services, WNPRC Genetics Services (MHC typing).
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