Analysis Of The Differences In Expressions Of Genes Between Murine Intestine Intraepithelial Lymphocytes And Splenic T Lymphocytes

Intestine intraepithelial lymphocytes (iIEL) are loosely organized infiltrated lymphocytes residing within the mucosa of the gut above the basal lamina of the epithelium. They play an important role in mucosal immunity and oral immune tolerance, etc. There are many differnces between iIEL and conventional T cells in morphology, phenotypes, ontogeny and functions. We assume that such differences should base on structurely a group of unique molecules. Therefore, we compared the differences in expressions of genes between murine iIEL and splenic T lymphocytes with a two-step strategy, establishing the cDNA subtractive library of iIEL firstly, and analyzing the iIEL special genes.The first, the iIEL and splenic T lymphocytes were isolated by density gradient in percoll and nylon column respectively. Both of the purities were above 90 percent. We then established the cDNA subtractive library of iIEL via improved subtractive hybridization. The cDNA was ligated with a different adaptor. After two rounds of hybridization and suppression PCR amplification, the PCR products v/ere directly inserted into the plasmid vectors. The ESTs special for iIEL were screened by reverse Northern blot. We screened 50 ESTs with obviously difference in the cDNA subtractive library of iIEL. These ESTs were squenced and submitted to Genebank(BI993440, BQ630825～BQ630873). We analyzed the ESTs throught BLAST, and obtained 11 genes associated with iIEL. They were Toll-like receptor 4(TLR4), Collectin sub-family member 12, Tyrosine protein kinase JAK3, Interleukin-1 receptor-associated kinase 1 (IRAK-1), ras-like GTP-binding protein 6 (RAB6), Cystatin C precursor, X-linked...