Numerous data demonstrate that lymphoid tissues represent a major HIV reservoir and sites of virus replication. To gain more information about the phenotype of cells in which HIV replicates in the lymphoid tissue, we applied HIV RNA hybridization technique in combination with immunohistochemistry on 10 lymph nodes and 2 lymphoepithelial cysts (LEC) of the parotid gland. In addition, to determine whether or not follicular dendritic cells (FDC) are important targets for HIV, we also analysed isolated and sorted FDC from 4 lymph nodes. The tissues showed follicular hyperplasia and heavy RNA signals in the germinal centres (GC), where diffuse labelling in the light zone as well as single cells with > 50 silver grains were present. The extrafollicular parenchyma contained only a few productively infected cells. The diffuse labelling was intimately related to the FDC network. However, electron microscopic examination and in situ hybridization on isolated FDC did not provide convincing evidence for virus replication in these cells. Virus-producing cells in the GC were almost exclusively CD45RO+, CD57- T cells. Macrophages with HIV RNA were mainly found in LEC of the parotid gland. S-100+ interdigitating cells of the T-dependent zone were HIV RNA-. However, some large S-100+ cells in the squamous epithelium covering the LEC or lying subepithelial did express HIV RNA. These findings suggest that local factors (cytokines?) in the lymphoid tissue probably influence the expression of HIV.