Analysis of RNA transcripts for HLA class II genes in human small intestinal biopsies.
Studies of the expression of selected genes within the intestinal mucosa will provide important new information about physiologic pathological processes that effect mucosal growth, differentiation, and function. To study gene expression in the gut, we developed a method to obtain sufficient undegraded RNA from human endoscopic intestinal biopsy specimens for Northern and slot blot analysis. To verify the method, we examined the differential expression of HLA class II genes in small intestinal mucosa. Levels of RNA transcripts for HLA-DR, -DP, and -DQ alpha and beta chains were assessed in freshly isolated endoscopic intestinal mucosal biopsy specimens and compared with levels in Epstein-Barr virus transformed B cells from the same individuals. Sufficient undegraded cellular RNA with distinct 28S and 18S ribosomal bands could be obtained from as few as two 2-3 mm endoscopic biopsies. Using chain and locus specific cDNA probes, HLA-DR, -DP, and -DQ subregion genes were shown to be expressed in intestinal mucosa, with the relative magnitude of RNA transcripts being DR greater than DP greater than DQ. The same hierarchy of expression was seen for EBV-transformed B cell lines. This method, in conjunction with the polymerase chain reaction for amplifying specific RNA transcripts and in situ hybridisation methods for the cellular localisation of RNA transcripts, will enable studies on the regulation of gene expression in the intestinal mucosa.
Volk
B A
BA
Department of Medicine, University of California, San Diego, La Jolla 92093.
Brenner
D A
DA
Kagnoff
M F
MF
D001706:Biopsy; D004386:Duodenum / Q000032:analysis* / Q000473:pathology; D005802:Genes, MHC Class II; D006681:HLA-D Antigens / Q000235:genetics; D006682:HLA-DP Antigens / Q000235:genetics; D006684:HLA-DR Antigens / Q000235:genetics; D000949:Histocompatibility Antigens Class II; D006801:Humans; D007413:Intestinal Mucosa / Q000032:analysis* / Q000473:pathology; D012313:RNA / Q000032:analysis*
C416186:H-2O antigen; D006681:HLA-D Antigens; C046148:HLA-DO antigens; D006682:HLA-DP Antigens; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; D012313:RNA