早期乳腺癌患者化疗前后外周血T细胞TCRβ CDR3组库动态变化特征  

The Dynamic Characteristics of TCR β CDR3 Repertoire of Peripheral Blood T Cells Before and After Chemotherapy in the Early Stage of Breast Cancer Patients

作　　者：杨丽文[1] 孙素红[2] 张天[1] 段芳芳[1] 贺晓燕[1] 马锐[1] 马龙[1] 石彬[3] 姚新生[1] Yang Liwen;Sun Suhong;Zhang Tian;Duan Fangfang;He Xiaoyan;Ma Rui;Ma Long;Shi Bin;Yao Xinsheng(Department of Immunology, Research Center for Medicine ＆ Biology, Innovation ＆ Practice Base for Graduate Students Education, Zunyi Medical University, Zunyi 563003, China;Department of Breast Surgery, the First Affiliated Hospital of Zunyi Medical University, Zunyi 563003, China;Department of Laboratory Medicine, Zunyi Medical University, Zunyi 563003, China)

机构地区：[1]遵义医学院免疫学教研室贵州省免疫学研究生教育创新基地,遵义563003 [2]遵义医学院附属医院甲乳外科,遵义563003 [3]遵义医学院检验医学系,遵义563003

出　　处：《中国细胞生物学学报》2018年第6期945-954,共10页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81441048);贵州省科技厅联合基金(批准号:黔科合LH字[2014]7584)资助的课题~~

摘　　要：该文采用高通量测序(high-throughput sequencing,HTS)技术分析早期乳腺癌患者化疗前后外周血T细胞TCRβCDR3(T-cell receptor beta chain complementarity determining region 3)组库的组成及动态变化特征,初步探讨化疗对T细胞的可能影响及化疗前后患者T细胞的应答功能状态。以病理活检确诊为T1N0M0期乳腺癌患者为研究对象,选取进行6期TAC[多西他赛(docetaxel,T)、吡柔比星(pirarubicin,A)、环磷酰胺(cyclophosphamide,C)]化疗方案。化疗的3例志愿患者(P1、P2、P3)分别于化疗前、第3期化疗后、第6期化疗后采集外周血分选出单个核细胞(peripheral blood mononuclear cells,PBMCs),提取其总DNA;利用多重PCR建立TCRβCDR3组库进行HTS;采用Immuno SEQ和IMGT-High-V-quest系统对TCRβCDR3序列进行组成和特征分析。结果表明:(1)P1、P2、P3在第3期化疗后的CDR3组库多样性[用反辛普森指数(1/DS)表示]相比化疗前均升高;第6期化疗后,P1和P3相比第3期化疗后CDR3组库多样性降低,但P2相比第3期化疗后CDR3组库多样性升高;(2)P1和P3在第3期化疗后CDR3组库高频克隆(大于1%)较化疗前出现降低,而第6期化疗后CDR3组库高频克隆相比第3期化疗后明显升高;P2第3期化疗后和化疗前相比CDR3组库高频克隆无变化,但第6期化疗后明显降低;(3)P1和P2化疗前后出现部分TRBV基因取用丢失与重新取用,P3的TRBV基因取用未发生明显改变;P2和P3化疗后部分TRBV与TRBJ优势配对消失。早期乳腺癌患者TAC化疗的前、中、后期,其外周血T细胞TCRβCDR3组库表现出个体化的多样性和特征性改变,可能与TAC对个体T细胞的抑制或杀伤相关。监测CDR3组库的组成及动态变化可为化疗对T细胞的影响及化疗前后"个体化"的T细胞免疫状态评估提供基础。Using high-throughput sequencing（HTS） technology, analysis of the dynamic change characteristics and composition of T cell of T-cell receptor beta chain complementarity determining region 3（TCR β CDR3） repertoire in peripheral blood before and after chemotherapy with breast cancer patients in the early period, preliminary discussion on the possible effect of chemotherapy on T cells and T cell response function status before and after chemotherapy patients. The pathological biopsy diagnosed patients with T1 N0 M0 breast cancer as the research object, a six periods TAC（T： docetaxel; A： pirarubicin; C： cyclophosphamide）chemotherapy in three volunteers patients（P1, P2, P3）, before chemotherapy, after the third periods of chemotherapy and the sixth periods of chemotherapy, then collected the peripheral blood and separated the PBMC, respectively, and extracted genomic DNA. Using the multiple PCR to construct human TCR β CDR3 repertoire and HTS, then using Immuno SEQ and IMGT-High-V-QUEST analysis the composition and characteristics of TCR β CDR3 sequences. The results suggest that after the third period of treatment, CDR3 repertoire diversity（the inverse Simpson index） was higher than before chemotherapy in P1, P2, P3. After the six period of treatment, compared with the third period of treatment, CDR3 repertoire diversity decreased in P1 and P3, but increased in P2. After the third period of treatment, in P1 and P3 patients, CDR3 repertoire highfrequency clones（greater than 1%） lower than the chemotherapy before. And after the sixth period of treatment, CDR3 repertoire high-frequency clones increased significantly compared with the third treatment. In P2 patient, there was no changes in the high-frequency clones in CDR3 repertoire after the third period of treatment compared with before chemotherapy, but after the sixth period of treatment, it was significantly reduced. The partial TRBVgene usage was lost and reused before and after chemotherapy in P1 and P2, and the TRBV gene usage of

关 键 词：高通量测序技术 T细胞CDR3组库 乳腺癌 化疗 

分 类 号：R737.9[医药卫生—肿瘤]