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Objectives:

HIV-1-specific CD8 T cells are considered to be critical in anti-HIV responses. It is important to quantify these cells and to determine their antigenic targets. Here quantification of interferon (IFN)-g secreting, virus-specific cells was achieved with an enzyme linked immuno spot (ELISPOT) assay.

Methods:

Peripheral blood mononuclear cells (PBMC) were infected with recombinant vaccinia vectors expressing HIV-1 genes (gag, pol, env or nef) and added to wells precoated with anti-IFN-g monoclonal antibodies. Spot forming cells (SFC), ie antigen-specific T cells were detected 24 h later by the addition of biotinylated anti-IFN-g monoclonal antibodies, followed by avidin-bound biotinylated horseradish peroxidase.

Results:

In a cohort of 19 patients, of whom 15 were on highly active antiretroviral therapy, 18 had primed T cells directed against one or more HIV-1 antigens (P< 0.0001). Pol-specific T cells …