乳腺癌患者外周血CD4^+ CD25^+ CD127^(dim/-) Tregs细胞TCRβ链CDR3谱型漂移特征初步探讨  

Preliminary study on drift characteristics of CD4^+ CD25^+ CD127^(dim/-) regulatory T cells TCR β chain CDR3 spectrum in peripheral blood of breast cancer patients

作　　者：明亚芳[1] 杨伟明[1] 任宇[1] 姚新生[2] 杨雪峰[1] 

机构地区：[1]遵义医学院附属医院,贵州遵义563003 [2]遵义医学院免疫学教研室

出　　处：《中国妇幼保健》2015年第34期6134-6138,共5页Maternal and Child Health Care of China

基　　金：贵州省教育厅重点项目〔黔教科(20090110号)〕;教育部新世纪人才项目〔NCET-10-0095〕

摘　　要：目的探讨乳腺癌患者外周血单个核细胞中CD4^+CD25^+CD127^(dim/-)Tregs细胞TCRB链CDR3的基因,氨基酸序列及初步结构。方法采用免疫磁珠细胞分选技术,分选出10例乳腺癌患者及5例正常患者外周血中的CD4^+CD25^+CD127^(dim/-)Tregs细胞,流式细胞术检测分选纯度;采用毛细吸管电泳法分析TCRB链CDR3优势利用情况,对单克隆增生的家族进行基因测序。结果 5例正常CD4^+CD25^+CD127^(dim/-)Tregs细胞TCR BV CDR3长度和多态性表达频率不一致,大部分呈高斯分布;10例乳腺癌患者CD4^+CD25^+CD127^(dim/-)Tregs细胞TCR BV CDR3家族长度和多态性表达频率不一致,乳腺癌患者外周血CD4^+CD25^+CD127^(dim/-)Tregs细胞TCR BV CDR3区谱型异常率明显大于正常人(χ~2=32.345,P<0.01),TCRBV22、TCRBV15、TCRBV16 3个家族被多例患者限制性取用,出现优势利用,单克隆增生的家族进行基因测序发现,病例1 TCR BV11和病例2 TCR BV4存在共同基序"ETQYF";病例2 TCR BV13.1和病例7 TCR BV15有共同基序"EQFF";病例2 TCR BV13.2和病例5 TCR BV22存在共同基序"TGELFF"。结论乳腺癌患者外周血CD4^+CD25^+CD127^(dim/-)Tregs细胞存在不同程度的克隆增生,部分TRBV家族被多例乳腺癌患者限制性取用并在免疫应答中发挥重要作用。多数TCR BV家族CDR3谱系存在单峰谱型,且部分单峰谱型的T细胞具有不同的CDR3序列,提示多个抗原表位参与了T细胞免疫应答。多数病例单峰谱型不同家族间存在共同基序,推测调节性T细胞可能识别抗原肽的相同部分。Objective To explore gene, amino acid sequence, and preliminary structure of CD4^＋ CD25^＋ CD127^dim/- regulatory T cells TCR β chain CDR3 in mononuclear cells of peripheral blood of breast cancer patients. Methods Immunomagnetic cell sorting technique was used to sort out CD4^＋ CD25^＋ CD127^dim/- regulatory T cells in peripheral blood of 10 breast cancer patients and five normal women, flow cytometry was used to detect the purity ; capillary electruphoresis was used to analyze the utilization of advantages of TCRβ chain CDR3, gene sequencing of monoclonal proliferation family was conducted. Results The family length and expression frequency of polymorphism of CD4^＋ CD25^＋ CD127^dim/- regulatory T cells TCR BV CDR3 of five normal women were not the same, showing a Gauss distribution; the family length and expression frequency of polymorphism of CD4^＋ CD25^＋ CD127^dim/-regulatory T ceils TCR BV CDR3 of ten breast cancer patients were not the same. The abnormal rate of CD4^＋ CD25^＋ CD127^dim/- regulatory T ceils TCR BV CDR3 spectrum in peripheral blood of breast cancer patients was statistically significantly higher than that of normal women （Х^2 = 32. 345, P〈0. 01 ）, TCRBV22, TCRBV15, and TCRBV16 families were used restrictively in many patients, the advantage of usage appeared, gene sequencing of monoclonal proliferation family showed that case 1 （TCR BV11） and case 2 （TCR BV4） had a common motif ＂EQFF＂ ; case 2 （TCR BV13.1 ） and case 7 （TCR BV15 ） had a common motif ＂EQFF＂ ; case 2 （TCR BV13.2） and case 5 （TCR BV22） had a common motif ＂TGELFF＂. Conclusion CD4^＋ CD25^＋ CD127^dim/- regulatory T cells in peripheral blood of breast cancer patients have different degrees of clonal proliferation, partial TRBV families are used restrictively by many breast caneer patients and play important roles in immune response. Most of TCR BV family CDR3 lineages are single peak spectral type, and some single peak spectral type T cells have different CDR 3 series,

关 键 词：乳腺癌 调节性T细胞 互补决定区3 

分 类 号：R737.9[医药卫生—肿瘤]