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SUMMARY: We used three-colour cytometry to analyse intracellular cytokine production in activated whole blood cultures derived from patients with HIV-1 infection. We assessed mitogen-induced IL-2, IL-4 and IFN-[gamma] production from T cells as possible markers of immune dysfunction. The percentages of T cells staining for IL-2 were significantly reduced in stimulated cultures from HIV individuals relative to normal controls(P<0[middle dot]0001); this reduction was observed in both the CD4 and the CD8 subsets. IL-2 production was significantly reduced in CD4 T cells from HIV individuals clinically classified as symptomatics compared with HIV asymptomatics(P<0[middle dot]001); in addition, production of IL-2 inversely correlated with viral load (r2=0[middle dot]832). On the other hand, HIV individuals showed significantly more T cells staining positive for IFN-[gamma] (P<0[middle dot]0001); subset analysis identified these T cells as CD8 . Increased IFN-[gamma] production in the CD8 T cell subset of HIV individuals correlated neither with clinical status nor with plasma viral load. IL-4 staining in activated T cells was low (<5%) and no differences were observed between HIV and control groups. Three-colour FACS analysis of whole blood provides a sensitive, rapid and relatively easy means to detect cytokine profiles within T cell subpopulations. Only small volumes of blood are required (0[middle dot]5 ml), since there is no need for cell isolation, making it more practical than ELISA or reverse transcriptase-polymerase chain reaction (RT-PCR) for the analysis of immune function in HIV individuals. This technique could therefore play a role in mapping the dynamics and extent of immune recovery in AIDS patients undergoing triple combination therapy.

(C) 1998 Blackwell Science Ltd.