Regulation of TNFalpha and TNFbeta gene expression in inflammatory bowel disease

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Tumor necrosis factor alpha (TNFalpha) and TNFbeta are cytokines with similar immunoregulatory effects binding to their target cells via TNF receptors. To clarify the regulation of TNFalpha and TNFbeta in the mucosal lesions of IBD patients, we examined levels of TNFalpha and TNFbeta mRNA transcripts and these peptide production in the mucosa.METHODS : 1)Intestinal specimens were obtained through an endoscopy or at operation, and total RNA was extracted by a GTP method. RNA was reverse transcribed with M-MLV-RT into cDNA. TNFalpha and TNFbeta mRNA were quantitated using linear PCR methods. As internal controls, cDNA was amplified by PCR with primers for beta-actin, T-cell receptor alpha, ferritin heavy chain. 2)Other mucosa from patients were cultured for 1,2 hours with LPS (10mug/ml) and tRNA in the mucosa was extracted.RESULTS & CONCLUSIONS : The level of TNFalpha mRN was 32-64 fold increased and TNFbeta mRNA was 4-16 fold increased in inflamed mucosa of IBD patients compared to mucosa of control donors. TNFalpha mRNA was 4-8 fold increased and TNFbeta mRNA was not increased in LPS primed IBD mucosa. TNFalpha is an important mediator in response to endotoxin in IBD.