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目的从新的角度探讨慢性乙型肝炎(CHB)炎症活动期外周血T淋巴细胞克隆性增生情况,了解T淋巴细胞免疫应答在CHB发病机制中的状态及作用。方法利用逆转录聚合酶链反应(RT-PCR) 扩增T淋巴细胞受体(TCR)β链V区基因各家族(BV),并采用免疫指纹技术(immunoscope)对4名健康献血员及8例处于炎症活动期CHB患者外周血TCR BV家族基因的优势利用情况及克隆性增生T淋巴细胞β链互补决定区3(CDR3)序列进行分析。结果 4名健康献血员外周血T淋巴细胞TCR BV家族CDR3谱型均呈正态分布,而8例CHB患者均出现一个或多个TCR BV家族单克隆或寡克隆性增生。对克隆性增生T淋巴细胞β链CDR3区序列测定证实存在不同的CDR3序列。结论 CHB活动期外周血T淋巴细胞存在明显克隆性增生,进一步提示T淋巴细胞免疫应答可能参与CHB的发病过程,且可能有多个病毒抗原表位参与了细胞免疫应答。
Objective To explore the clonal proliferation of peripheral T lymphocytes in chronic hepatitis B (CHB) from a new perspective and to understand the status and role of T lymphocyte immune responses in the pathogenesis of CHB. Methods The families of BV gene of TCR β chain were amplified by reverse transcription polymerase chain reaction (RT-PCR), and four healthy blood donors and 8 Cases of inflammatory activity of CHB patients with peripheral blood TCR BV family advantage of utilization and clonality of T lymphocyte β chain complementarity determining region 3 (CDR3) sequence analysis. Results The TCR BV family CDR3 profiles of peripheral blood T lymphocytes from four healthy donors showed a normal distribution. One or more TCR BV family monoclonal or oligoclonal hyperplasia occurred in all 8 CHB patients. Sequencing of the β chain CDR3 region of clonogenic T lymphocytes confirmed the presence of different CDR3 sequences. Conclusion There are obvious clonal hyperplasia of T lymphocytes in the peripheral blood of patients with CHB, which further suggests that the T lymphocyte immune response may be involved in the pathogenesis of CHB and there may be multiple viral epitopes involved in the cellular immune response.