Extraction of immune and inflammatory cells from human lung parenchyma: evaluation of an enzymatic digestion procedure.
The inflammatory and immune cell populations of the human lung parenchyma have not been characterized in detail. This report describes a novel and efficient procedure for their extraction. Histologically normal human lung tissue samples from pneumonectomy specimens were sliced to 0.5 mm, and digested in collagenase/DNAse. Viable mononuclear cell yields ranged from 15-48 X 10(6)/g, and were markedly in excess of reported methods employing mechanical tissue disruption, which normally yield populations containing almost exclusively macrophages. The lung digest population was examined by flow cytometry using monoclonal antibodies against cell surface receptors, and found to comprise up to 40% T lymphocytes, 10% B lymphocytes and 30% macrophages, contaminated by less than 1% peripheral blood cells. Based upon these figures, the recoverable lung parenchymal lymphoid cell pool appears considerably larger than previously recognized, being of the same order as the peripheral blood pool. Initial functional studies suggest that such cellular activities as antigen-specific T cell proliferation, antigen-presentation, interleukin 1 production and natural killer cell activity survive the extraction process, and controlled enzymatic digestion experiments with peripheral blood cells indicate that the degree of enzyme-mediated damage to these functions and to cell-surface structures, was minimal. The extraction method thus appears suitable for studying the types and functions of human parenchymal lung cells in health and disease.
D000368:Aged; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D002452:Cell Count; D003584:Cytological Techniques; D003851:Deoxyribonucleases / Q000494:pharmacology; D006168:Guinea Pigs; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology; D008168:Lung / Q000166:cytology* / Q000276:immunology / Q000648:ultrastructure; D008213:Lymphocyte Activation; D051379:Mice; D003012:Microbial Collagenase / Q000494:pharmacology; D008854:Microscopy, Electron; D008875:Middle Aged; D051381:Rats; D013601:T-Lymphocytes / Q000276:immunology
D000954:Antigens, Surface; D003851:Deoxyribonucleases; D003012:Microbial Collagenase