T-lymphocyte activation was investigated in peripheral blood and bronchoalveolar lavage (BAL) of four patients with hypersensitivity pneumonitis. The study was performed by flow cytometry with the use of immunofluorescence labeling with monoclonal antibodies to lymphocyte differentiation or activation antigens. Simultaneous measurement of DNA and RNA content by acridine orange staining was used for cell-cycle analysis. The various cell types were identified by their light-scattering properties. T cell activation was demonstrated in the BAL of all patients by the presence of T cells (OKT3 positive) bearing class II histocompatibility antigen (HLA-DR) and activated T cell markers (MLR 1-3). Lymphocyte proliferation was evidenced in BAL but not in blood of patients by an increased percentage of cells in S + (G2 + M) phases. In addition, T-lymphocyte subsets analysis revealed no abnormalities in the blood and no major imbalance in the BAL despite a slightly increased OKT4/OKT8 ratio. The finding of T cell activation and lymphocyte proliferation in hypersensitivity pneumonitis alveolitis is consistent with the contribution of a local type IV immune reaction to the pathogenesis of this disease.