乳腺癌患者化疗前后外周血B细胞BCR H-CDR3受体库动态变化特征  

Dynamic characteristics of BCR H chain CDR3 repertoire of peripheral blood B cells before and after chemotherapy in the breast cancer patients

作　　者：段芳芳[1] 孙素红[2] 张天[1] 李月红[1] 姚新生[1] DUAN Fang-fang;SUN Su-hong;ZHANG Tian;LI Yue-hong;YAO Xin-sheng(Department of Immunology,Research Center for Medicine ＆ Biology,Innovation ＆ Practice Base for Graduate Students Education,Zunyi Medical University,Zunyi 563000,China;Department of Breast Surgery,the First Affiliated Hospital of Zunyi Medical University,Zunyi 563000,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563000 [2]遵义医学院附属医院甲乳外科,遵义563000

出　　处：《现代免疫学》2018年第4期301-309,共9页Current Immunology

基　　金：贵州省科技厅联合基金,黔科合LH字【2014】7584号; 国家自然科学基金(81441048)

摘　　要：本文利用高通量测序(high-throughput sequencing,HTS)技术对早期乳腺癌患者化疗前后,随着WBC总数及淋巴细胞的变化对外周血B细胞BCR H-CDR3受体库进行分析,探讨化疗对其的影响。分别采集和分离3例进行TAC化疗的乳腺癌患者在化疗前、中、后的PBMC,提取其总DNA并采用多重PCR建库和制备BCR H-CDR3受体库后Illumina检测CDR3序列并分析。结果发现:1.TAC化疗前患者WBC、淋巴细胞绝对值均正常,而化疗中、后WBC、淋巴细胞绝对值虽仍正常,但较化疗前下降;2.P2化疗中较化疗前受体库多样性有所下降,化疗后较化疗中稍升高,但仍明显低于化疗前;P1、P3化疗中多样性都比化疗前高;化疗后,P1较化疗中降低,但高于化疗前,而P3化疗后较化疗中明显升高;3.3例患者化疗后出现不同程度IGHV取用丢失。由此提示:1.化疗后随着WBC、淋巴细胞绝对值的下降,P2乳腺癌患者在化疗中及化疗后受体库的多样性在整体水平上较化疗前有所下降,考虑是TAC对受体库的多样性造成损伤;而P1、P3化疗中与化疗后B细胞受体库多样性整体上都较化疗前高,可能是机体对化疗药物产生的应激反应不一致,TAC杀伤了高频克隆的B细胞,间接增加了CDR3受体库的多样性;2.3例早期乳腺癌患者化疗后出现不同程度IGHV取用丢失,提示化疗可能对B细胞IGHV表达和取用产生了一定的影响,并且IGHV-IGHJ家族取用和配对在不同时间点存在优势配对(>0.03)现象,可能与乳腺癌相关抗原诱导有关;3.HTS检测化疗前后B细胞H-CDR3受体库变化可为揭示化疗对患者B细胞应答的影响及B细胞的免疫状态提供辅助。We aimed to explore the impact of chemotherapy on CDR3 repertoire of BCR H chain.High-throughput sequencing technique was used to analyze chemotherapy impact on BCR H CDR3 repertoire of peripheral blood B cells in early breast cancer patients.Three patients with breast cancer who underwent TAC were enrolled and PBMCs were collected before,during and after chemotherapy.Genomic DNA was extracted and BCR H chain CDR3 repertoire was constructed using multiple PCR and Illumina sequencing of CDR3 was performed.The results were as follows：1.The WBC count and the absolute value of lymphocytes of the 3 patients remained in the normal range,although both were lower than before chemotherapy.2.In patient 2,chemotherapy reduced BCR repertoire which slightly increased after treatment but remained lower than that before chemotherapy.In patient 1 and patient 3,the BCR H chain repertoire was enhanced with the treatment.3.All the 3 patients had a loss of IGHVusage in different degree.Overall,these findings suggest that B cell IGHVexpression and usage were influenced by chemotherapy and there is a dominant pairing（P〉0.03）of the IGHV-IGHJgenes pair at different time points.Detection of B cell H-CDR3 repertoire before and after chemotherapy by using HTS could be helpful to reveal the effect of chemotherapy on the response of B cells and to understand the immune status of B cells.

关 键 词：高通量测序技术 乳腺癌 化疗 BCR CDR3受体库 

分 类 号：R392.12[医药卫生—免疫学]