ISOLATION OF FUNCTIONALLY ACTIVE INTRAEPITHELIAL LYMPHOCYTES AND ENTEROCYTES FROM HUMAN SMALL AND LARGE-INTESTINE

被引:66
作者
LUNDQVIST, C
HAMMARSTROM, ML
ATHLIN, L
HAMMARSTROM, S
机构
[1] UMEA UNIV,DEPT IMMUNOL,S-90187 UMEA,SWEDEN
[2] UMEA UNIV,DEPT SURG,S-90187 UMEA,SWEDEN
关键词
INTRAEPITHELIAL LYMPHOCYTE; ENTEROCYTE; ISOLATION PROCEDURE; (HUMAN);
D O I
10.1016/0022-1759(92)90147-L
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A mild purification method has been developed for the isolation of human intraepithelial lymphocytes (IEL) and enterocytes from the same individual. The isolation procedure includes mechanical disruption of the mucosal layer, treatment with reducing agent and sedimentation followed by Percoll gradient centrifugation. Finally, epithelial cells are removed from the IEL fraction using magnetic beads coated with the anti-epithelial antigen monoclonal antibody (mAb) BerEP4. Leucocytes are removed from the enterocyte fraction using magnetic beads coated with mAbs directcd against common leucocyte antigen (CD45). Using this procedure IEL and enterocytes have been isolated from apparently normal jejunal, ileal and colonic tissue specimens. Recoveries of IEL were 7 x 10(5), 4 x 10(5) and 1 x 10(5)/cm2 mucosa from jejunum, ileum and colon respectively. 1-2 x 10(6) enterocytes/cm2 mucosa were recovered from small intestine while the corresponding value for colonic biopsies was approximately 2 x 10(5) enterocytes/cm2. The IEL fraction was pure as judged by the low percentages of B cells, macrophages and BerEP4 positive cells (< 4%) present in the purified fraction. The enterocyte fraction contained < 2% CD45+ cells. The two cell fractions were viable and expanded in vitro. Enterocytes expanded spontaneously while IEL required initial stimulation with mitogens. The isolation procedure described here will make it possible to study the function of human IEL, interactions between IEL and enterocytes and the role of both cell types in local immunity.
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页码:253 / 263
页数:11
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