Mechanisms of HIV non-progression; robust and sustained CD4+ T-cell proliferative responses to p24 antigen correlate with control of viraemia and lack of disease progression after long-term transfusion-acquired HIV-1 infection

Dyer, Wayne B; Zaunders, John J; Yuan, Fang Fang; Wang, Bin; Learmont, Jennifer C; Geczy, Andrew F; Saksena, Nitin K; McPhee, Dale A; Gorry, Paul R; Sullivan, John S

The response of CD8+ T cells to HIV antigen was measured by IFNγ ELISPOT, using pre-coated ELISPOT kits according to the manufacture's protocol (Mabtech, Mosman, Australia). Firstly, the response to whole HIV proteins was determined in response to antigen presented by autologous B lymphoblastoid cell lines infected for 18 hours with 5 pfu/cell recombinant Vaccinia expressing the HIV-1 IIIB env, gag, pol, or nef genes (Therion Biologics, Cambridge, MA, USA), or E. coli lacZ as a control. Gag responses were further characterised using overlapping Gag peptides, firstly using a matrix of peptide pools, and then individual peptides for confirmation (full Gag peptide set; kindly provided by the NIH AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH).