Intraepithelial, lamina propria and Peyer's patch lymphocytes of the rat small intestine: isolation and characterization in terms of immunoglobulin markers and receptors for monoclonal antibodies.
Methods have been determined for the isolation, purification and subsequent characterization of separate populations of rat intestinal lymphoid cells, namely intraepithelial (IEL), lamina propria (LPL) and Peyer's patch lymphocytes (PPL). Dissociation of the epithelium from the basement membrane with subsequent release of IEL was achieved by citrate buffer incubation followed by vortex agitation. LPL were released from the remaining tissue by scraping, and PPL were similarly obtained. Some preparations of lamina propria were further subjected to collagenase digestion. After filtration and density gradient centrifugation, average yields of 220 x 10(4) IEL, 54 x 10(4) LPL and 220 x 10(4) PPL per gram of gut were obtained. Immunofluorescence characterization demonstrated that cells bearing the MRC OX8 (T-suppressor) marker predominated in IE1 (73%) and were present in lower concentrations in LPL (26%) and PPL (6%). Cells with the W3/25 (T-helper) marker accounted for a small proportion of each of the lymphocyte preparations. IE1 were unusual in containing a population of cells which were negative for the W3/13 marker for T cells, but were MRC OX8 positive. B lymphocytes were present in PPL (55%) and LPL (31%), but were virtually absent in IEL (less than 1%). Few plasma cells were observed. The techniques described will allow functional investigations to be made and lead to a better understanding of mucosal immunity.
D000818:Animals; D002469:Cell Separation; D003593:Cytoplasm / Q000276:immunology; D004229:Dithiothreitol / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000276:immunology; D005455:Fluorescent Antibody Technique; D007413:Intestinal Mucosa / Q000276:immunology; D007421:Intestine, Small / Q000276:immunology*; D008214:Lymphocytes / Q000032:analysis / Q000145:classification / Q000276:immunology*; D008221:Lymphoid Tissue / Q000166:cytology*; D010581:Peyer's Patches / Q000166:cytology* / Q000276:immunology; D051381:Rats; D011919:Rats, Inbred Strains; D011947:Receptors, Antigen, B-Cell; D011948:Receptors, Antigen, T-Cell / Q000276:immunology
D011947:Receptors, Antigen, B-Cell; D011948:Receptors, Antigen, T-Cell; D004229:Dithiothreitol