IMMUNOREGULATORY DEFECTS IN INFLAMMATORY BOWEL DISEASE

炎症性肠病的免疫调节缺陷

• 批准号: 3790738
• 负责人: W STROBER
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3790738
• 关键词: Crohn's disease; T cell receptor; autoantigens; human subject; immunopathology; immunoregulation; inflammatory bowel diseases; intestinal mucosa; messenger RNA; polymerase chain reaction; receptor expression; ulcerative colitis

Over the past several years we have examined the hypothesis that the inflammatory bowel diseases, ulcerative colitis and Crohn's disease, is due most fundamentally to an abnormal immune response to one or more mucosal antigens that are always present in the mucosal environment. During this period we investigated this possibility by assessing TCR-V-beta family repertoire expression in T cells in the lamina propria (LPMC) and peripheral blood (PBMC) of IBD patients and control individuals. Our strategy for making such determinations was based on a quantitative RT-PCR technique that has been developed in this laboratory. This technique involves first the construction of internal mRNA standards having the same primer sites as the mRNA to be quantitated but having a different size from the latter and second the co-reverse transcription and amplification of the standard mRNA. The amount of "unknown" TCR-V-beta mRNA is determined in this method from an equivalence point at which standard and unknown are transcribed and amplified with equal efficiency. In our initial studies we found using a qualitative RT-PCR technique that LPMC mRNA of IBD patients contained a full repertoire of 20 TCR-V-beta families and that IBD is not associated with gross deletion or expansion of any family. In further studies with quantitative RT-PCR of four frequently expressed TCR-V-beta families, we found that relative TCR-V-beta expression in LPMC + PBMC was quite different, probably reflecting the somewhat different antigen-drive giving rise to these cell populations. More importantly, we found that mean mRNA values for TCR-V-beta2, but not TCR-V-beta6,7 and 14 were lower in IBD patients than in control individuals. These results indicate a selective abnormality of TCR-V-beta2 expression in both forms of IBD. They are therefore consistent with the concept that a particular antigen or superantigen is important in disease pathogenesis.

在过去的几年里，我们已经研究了假设， 炎症性肠病，溃疡性结肠炎和克罗恩病， 最根本的原因是对一种或多种 粘膜抗原总是存在于粘膜环境中。 在此期间，我们通过评估 固有层中T细胞中TCR-V-β家族库表达 IBD患者和对照的LPMC和外周血（PBMC） 个体 我们做出这种决定的策略是基于一个 定量RT-PCR技术，已在本实验室开发。 该技术首先涉及构建内部mRNA标准品 具有与待定量的mRNA相同的引物位点，但具有 与后者大小不同，其次是共逆转录 和标准mRNA的扩增。 “未知数”的数量 TCR-V-β mRNA在该方法中从等效点测定， 其中标准和未知的转录和扩增是相同的 效率 在我们最初的研究中，我们发现使用定性RT-PCR IBD患者LPMC mRNA包含一个完整的库， 20个TCR-V-β家族，IBD与总体缺失无关 或任何家庭的扩张。 在定量RT-PCR的进一步研究中 在四个频繁表达的TCR-V-β家族中，我们发现， LPMC + PBMC中的TCR-V-β表达有很大差异，可能是 这反映了产生这些细胞的有些不同的抗原驱动 人口。 更重要的是，我们发现， TCR-V-β 2，而不是TCR-V-β 6，7和14在IBD患者中低于 控制个体。 这些结果表明， TCR-V-β 2在两种IBD中的表达。 因此它们 这与特定抗原或超抗原 在疾病发病机制中重要。