Longitudinal analysis in one individual. (A) Time courses of the surrogate markers are shown. CD4+ T cell count (left axis, white circle) and pVL (right axis, black circle) are plotted on the chart. Each arrow indicates the earliest time point when the sequence above the arrow was observed. (B) IFN-γ ELISpot assay was performed by using serially diluted peptide set and expanded PBMCs. Two time points were tested for the assay. At the earlier point, 2000/9/4, the patient harbored wild type sequence around the Nef126-10/Nef134-10 region, and at the latter point, 2003/3/20, the sequence displayed 133T/135F. Mutants as well as wild type peptides of Nef126-10 or Nef134-10 epitopes were used in the assay. Each point represents the average SFU of duplicate wells after subtraction of background, and each error bar shows standard deviation.