Separation and Characterization of Lymphocytes from Rat Lung Parenchyma

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Abstract
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 × 106 cells per animal were recovered. On the basis of cy to chemical and morphologic criteria, the separated cells contained > 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 phytohemagglutin in and concanavalin A. Supernatants from lung lymphocytes stimulated with concana-valin A contained lymphokine activity that could be demonstrated in a leukocyte procoagulant activity assay.