摘要
目的 FQ-PCR溶解曲线法初步分析1例弥漫性大B淋巴细胞瘤患者组织标本中T细胞TRAV和TRBV基因各家族CDR3谱序。方法提取4例淋巴结增生组织和1例弥漫性大B淋巴细胞瘤患者的mRNA,逆转录成cDNA,FQ-PCR扩增32个TRAV基因和24个TRBV基因的CDR3区,利用FQ-PCR技术了解其克隆性,溶解曲线单峰家族基因测序分析CDR3区基因和氨基酸序列。结果该例淋巴瘤患者淋巴结组织T细胞FQ-PCR溶解曲线分别在TRAV10、TRAV18和TRBV6、TRBV24家族中出现单峰表现,其CDR3区氨基酸序列为CAGANFGNEKLTFGTGTCR、CAPSFSGYSTLTFGKGTM、CASSERGQPQHFGDGTCR、CATTPAGEYYGYTFGSGT-CR,4例增生淋巴结组织FQ-PCR溶解曲线均为寡峰和多峰。结论该例淋巴瘤组织中FQ-PCR溶解曲线单峰家族的T细胞可能为高应答T细胞,本实验为进一步研究淋巴溜的免疫应答机制和个体化治疗提供了研究基础。
Objective To analyze the CDR3 sequence of TRAV and TRBV gene repertoire in tissue of a patient with diffuse large B cell lymphoma by using real-time fluorescence quantitative reverse transcription polymerase chain reaction（FQ-PCR） with DNA melting curve analysis.Methods Total RNA from tissue of a patient with diffuse large B cell lymphoma and 4 patients with lymph node hyperplasia were transcripted reversely into cDNA.The cDNA of 32 TRAV and 26 TRBV gene families were amplified by FQ-PCR.Using DNA melting curve analysis techniques determined the features of 34 TRAV and 26 TRBV gene families.According to the melting curve,we selected the PCR products of TRAV and TRBV families showing monoclonal expansion,and then analysis their CDR3 spectratype.Results The melting curve showed single peak in TRAV10、TRAV18 and TRBV6、TRBV24 families.And the amino acid sequences of CDR3 region were CAGANFGNEKLTFGTGTCR,CAPAFSGYSTLTFGKGTM,CASSERGQPQHFGDGTCR,CATTPAGEYYGYTFGSGTCR respectively.Conclusion The data will be helpful to study the mechanism of immune response and individualized immunotherapy of diffuse large B cell lymphoma.
出处
《遵义医学院学报》
2012年第2期98-103,共6页
Journal of Zunyi Medical University
基金
贵州省科技厅国际合作计划(NO:[2008]700105)
贵州省科技厅社会发展基金项目(NO:黔科合SY字[2009]3098)
贵州省自然科学基金项目(NO:[2007]2122)
关键词
弥漫性大B淋巴细胞瘤
TCR
CDR3
FQ-PCR溶解曲线分析技术
Diffuse large B cell lymphoma
T-cells receptor
Complementarity determining region 3
Real-time fluorescence quantitative reverse TCR anscription polymerase chain reaction（FQ-PCR）
