alpha-Defensins 1, 2, and 3 exert antiretroviral activity in vitro, but their role in controlling HIV-1 replication in vivo and the cells that produce them are controversial. This study sought to determine whether alpha-defensins are present in HIV-1-infected individuals' lymphoid tissues, the major site of HIV-1 replication, and to identify the cells that express them. alpha-Defensin expression was evaluated by immunostaining inguinal lymph node sections from 19 untreated HIV-1-infected individuals and 8 individuals at low risk or seronegative for HIV-1 infection. Percentages of tissue sections that stained positively for alpha-defensins were not significantly different between HIV-seropositive (median, 7.6%) and -seronegative (median, 5.5%) individuals. Conditions that could have produced lymph node inflammation were present in most seronegative subjects, and their lymph node weights correlated with alpha-defensin expression (Spearman rho = 0.833; P = 0.010). A median of 100% (range, 95%-100%) of alpha-defensin-expressing lymph node cells from 8 subjects coexpressed the granulocyte marker, CD15. CD15 and alpha-defensin staining correlated (Spearman rho = 0.622; P < 0.001). These data suggest that alpha-defensins within lymphoid tissue are expressed by granulocytes and are prevalent in HIV-1-seronegative individuals with inflammatory processes as well as HIV-1-infected individuals. The role of alpha-defensins in controlling HIV-1 replication merits further investigation.