中国汉族人药物性皮炎相关T细胞受体的识别和功能验证

With more and more available new drugs , the morbility and mortality of drug-induced dermatitis increase dramatically. However,there are few efficient in vitro diagnostic methods.Studies indicate that the interaction among drug,peptides,HLA,T-cell receptor(TCR) play a pivotal role in the pathogenesis of drug-induced dermatitis. Our previous studies found that the detection of HLA alleles could to some extent reduce the prevalence of drug-induced dermatitis. A recent study in Taiwan identified a specific T-cell clone which can act as a biomarker for carbamazepine-induced SJS/TEN.However,for most other causative drugs,little is known regarding whether and how particular TCRs participate in the recognition of small drug compounds presented by HLA in T cell-mediated drug-induced dermatitis. Therefore, we try to identify the potential particular TCR by spectratyping the third complementarity-determing region(CDR3) of TCR and flow cytometry, validate its function in vitro by molecular dockings. With analyzing of the expression of potential TCR and cytotoxic cytokines expression in patients' serum and tissue, we aim to figure out the correlation among HLA, TCR and clinic phenotypes of patients. This study will provide new biomarker besides HLA allesles for the prevention and diagnosis of drug-induced dermatitis,compensating the shortage of in vitro screening tests at present.It is meaningful for the safety of invidualized medications.

随着疾病谱的改变和药物的开发使用，药物性皮炎(药疹)的发病率逐年增高，对药疹的预防及诊断缺乏有效手段。我们的前期研究发现,检测药疹相关的人类白细胞抗原（HLA）等位基因可某种程度上避免药疹的发生,但一些药物耐受者同样具有相关基因的表达、携带相关基因的患者临床严重程度不同，说明除HLA分子外尚有其他因素参与药疹的发生。台湾地区的研究发现卡马西平引起的重症药疹与单克隆T细胞受体（TCR）相关，但针对临床常见的其他致敏药物诱发药疹的研究未见报道。为此，我们在进一步挖掘及验证中国汉族人药物性皮炎易感基因的基础上，应用TCR系谱分析及流式细胞分析等方法识别药疹的风险TCR，通过阻断风险TCR来验证TCR的功能；结合外周血及组织中细胞毒性细胞因子的检测，明确HLA、TCR和药疹临床表型之间的关系。本实验结果可弥补单纯HLA基因筛查在药疹预防中的不足，提高药物开发和使用的安全性。

随着疾病谱的改变和药物的开发使用，药物性皮炎(药疹)的发病率逐年增高.对药疹的预防及诊断缺乏有效手段。我们的前期研究发现,检测药疹相关的人类白细胞抗原（HLA）等位基因可某种程度上避免药疹的发生,但一些药物耐受者同样具有相关等位基因的表达、携带相关等位基因的患者临床严重程度不同，说明除HLA分子外可能尚有其他因素参与了药疹的发生。因此，我们首先进一步挖掘药疹的风险HLA等位基因，包括醋甲唑胺、克林霉素、克拉霉素、破伤风抗毒和血塞通所致的药疹；应用高通量测序技术检测药物患者的TCR免疫组库偏移情况及其与病情的相关性，并识别药物特异性的TCR；检测药疹患者外周血及组织中细胞毒性细胞因子的表达，并分析其与临床表现的相关性；分析药疹患者病毒感染情况及其与患者预后的相关性，从而进一步明确药物性皮炎的发病机理。