Isolation and purification of human large bowel mucosal lymphoid cells: effect of separation technique on functional characteristics

• Published in: Gut Vol. 20; no. 12; pp. 1037 - 1046
• Main Authors: Bland, P W, Richens, E R, Britton, D C, Lloyd, J V
• Format: Journal Article
• Language: English
• Published: England BMJ Publishing Group Ltd and British Society of Gastroenterology 01.12.1979; BMJ Publishing Group LTD
• Subjects: Antibody-Dependent Cell Cytotoxicity; Cell Separation - methods; Centrifugation, Density Gradient; Humans; Intestinal Mucosa - cytology; Intestinal Mucosa - immunology; Intestine, Large - cytology; Intestine, Large - immunology; Leukocyte Count; Lymphocytes - analysis; Lymphocytes - immunology; Microbial Collagenase; Prostaglandins - analysis
• ISSN: 0017-5749; 1468-3288; 1458-3288
• DOI: 10.1136/gut.20.12.1037

Human large bowel lamina propria lymphoid cells have been isolated using both mechanical and enzymatic techniques. Their separation from other cell types after isolation was effected with greater efficiency by sedimentation on isokinetic gradients than by filtration through glass bead columns. After being purified, the capacity of the lamina propria lymphocytes to function in vitro as effector cells in antibody-dependent cellular cytotoxicity was determined. Mechanical distruption of the mucosa gave low yields of lymphoid cells, which lacked the capacity for cytotoxicity. Enzymatic digestion of mucosal tissue, by comparison, yielded large numbers of viable lymphoid cells which retained a significant level of cytotoxic activity. Investigation revealed that mechanical homogenisation stimulated the synthesis of prostaglandin E2, and inhibitor studies showed that this mediator was responsible for the lack of cytotoxic activity in mechanically-liberated lymphocytes.