Flow cytometric analysis of surface major histocompatibility complex class II expression on human epithelial cells prepared from small intestinal biopsies.
A technique for preparing viable, single cell suspensions of the epithelial layer of small intestinal tissue obtained endoscopically is described. Constant agitation of four biopsies for 60 min in the presence of chelating and reducing agents gave yields of 1.2-6.7 x 10(6) cells, of which 11-30% were intraepithelial lymphocytes (IEL). Passage through a nylon wool column removed dead cells. This preparation was suitable for flow cytometric analysis. Using this technique, surface MHC class II molecule expression was studied in 14 patients with normal small intestinal mucosa. Fluorescence labelling of these cells showed strong HLA-DR expression by epithelial cells (EC), DP was expressed less strongly, while little DQ expression could be detected. This technique demonstrates that small intestinal biopsies taken during routine endoscopy can yield adequate numbers of viable epithelial cells to perform flow cytometric analysis.
Madrigal
L
L
Department of Immunology, Trinity College Medical School, St. James's Hospital, Dublin, Ireland.
Lynch
S
S
Feighery
C
C
Weir
D
D
Kelleher
D
D
O'Farrelly
C
C
D000293:Adolescent; D000328:Adult; D000368:Aged; D001706:Biopsy; D002470:Cell Survival; D002478:Cells, Cultured; D004386:Duodenum / Q000166:cytology / Q000276:immunology*; D004847:Epithelial Cells; D004848:Epithelium / Q000276:immunology; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006682:HLA-DP Antigens / Q000096:biosynthesis; D006683:HLA-DQ Antigens / Q000096:biosynthesis; D006684:HLA-DR Antigens / Q000096:biosynthesis; D000949:Histocompatibility Antigens Class II / Q000096:biosynthesis*; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology*; D008297:Male; D008875:Middle Aged
D006682:HLA-DP Antigens; D006683:HLA-DQ Antigens; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II