Similar findings were obtained in an analysis restricted to sequences derived from plasma; the number of PNLGS in V1V2 increased with calendar year of sampling (β = 0.05 increase per year; p = 0.001), was increased in those with stage 3 infection (β = 1.14, p = 0.001) and was decreased in those with CD4 counts <200 cells/ml (β = −0.84, p = 0.04 compared to CD4 counts >500). This evidence concerns the gene CD4 and infection.