In tissue culture models of chronic human immunodeficiency virus type 1 (HIV-1) infection, cytokines such as tumor necrosis factor alpha (TNF-α) activate viral gene expression. We sought evidence that TNF-α might similarly regulate viral gene expression in vivo in the major lymphoid tissue (LT) reservoir. We used in situ hybridization, quantitative image analysis, and double-label techniques to compare cytokine and HIV-1 RNA levels in sections of tonsil and lymph node tissues obtained from individuals in early and later stages of HIV-1 infection. The levels of TNF-α gene expression in LT from HIV-1-infected and uninfected individuals were indistinguishable, and we found no correlation between TNF-α gene expression in LT and the level of HIV-1 gent expression in LT. There is thus little evidence that in vivo TNF- α significantly influences HIV production in LT.