Separation and Characterization of Lymphocytes from Rat Lung Parenchyma

Rat pulmonary parenchymal tissue was disaggregated into a single cell suspension by treatment with collagenase. A cell population enriched for lung lymphocytes was separated by depletion of adherent cells on a Sephadex G-10 column: 16.7 +/- 2.7 X 10(6) cells per animal were recovered. On the basis of cytochemical and morphologic criteria, the separated cells contained greater than 80% lymphocytes, with a viability of 80-90%. The distribution of lymphocyte subpopulations was determined by indirect immunoperoxidase staining with appropriate monoclonal antibodies. Separated lung lymphocytes exhibited a proliferative response in vitro to phytohemagglutinin and concanavalin A. Supernatants from lung lymphocytes stimulated with concanavalin A contained lymphokine activity that could be demonstrated in a leukocyte procoagulant activity assay.