异基因造血干细胞移植受者T细胞受体β链CDR3谱型表达与巨细胞病毒激活  

T cell receptor β-chain CDR3 spectratyping and cytomegalovirus activation in allogeneic hematopoietic stem cell transplant recipients

作　　者：吴志华 荆敏 梁韩英 杨镕 黄雅萍 陈晓明 胡建华 范骏 

机构地区：[1]浙江大学医学院附属第一医院传染病诊治国家重点实验室 [2]浙江省立同德医院检验科

出　　处：《浙江大学学报（医学版）》2016年第5期515-521,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金(LY14H190002);浙江省医药卫生科技计划(2013KYB084)

摘　　要：目的：探讨T细胞受体（TCR）β链可变区域（BV）的互补决定区（CDR3）谱型表达与异基因造血干细胞移植（HSCT）受者CMV激活的关系。方法：采用荧光定量PCR熔解曲线技术，扩增测序7例HSCT受者和3名健康对照者外周血单个核细胞的TCRBV家族CDR3谱型；采用免疫组织化学法检测外周血白细胞中的CMV—pp65抗原；应用ELISA法检测HSCT受者血清中的CMV—IgM。分析TCRBV家族CDR3表达与CMV激活的相关性。结果：3名健康对照者24个TCRBV家族均表达。7例HSCT受者术后TCRBV家族CDR3测序结果显示为BV9、BV11、BV17、BV20等BV家族序列；TCRBV9含“QVRGGTDTQ”，TCRBV11含“VATDEQ”和“LGDEQ”，TCRBV17含“IGQGNTEA”，TCRBV20含“VGLAANEQ”等共有氨基酸序列。抗原检测结果显示：术后3个月7例受者中有5例受者抗原血症阳性，CMV—pp65阳性细胞数为（2—15）个／5×10^4白细胞。抗体检测结果显示：术后3个月7例受者中3例受者CMV-IgM阳性。TCRBV家族CDR3的表达在CMV抗原血症阳性受者与抗原血症阴性受者之间差异无统计学意义（均P〉0．05）；但TCRBV11家族的表达在CMV-IgM阳性受者与CMV-IgM阴性受者之间差异有统计学意义（P〈0．05）。结论：HSCT受者在T细胞免疫应答中有特定的TCRBV家族CDR3谱型，其中TCRBV11的表达可能与CMV激活有关。Objective： To explore the association between T-cell receptor beta variable （TCR BV） complementarity determining region 3 （CDR3） spectratyping and CMV activation in the recipients of allogeneic hematopoietic stem cell transplantation （HSCT）. Methods： Fluorescence quantitative PCR melting curve analysis was used to sequence 24 TCR BV families in 7 HSCT recipients and 3 healthy controls. CMV-pp65 antigenemia was measured by immunohistochemical staining. Plasma IgM specific for CMV was identified using ELISA. Relationship between TCR BV families and CMV activation was statistically analyzed. Results： Twenty-four TCR BV families were expressed in 3 healthy controls, while TCR BV CDR3 sequencing results in 7 recipients turned out to be BV9, BV11, BV17, BV20 and so on. Amino acid sequence features were as follows： TCR BV9 contained ＂ QVRGG＇ ＂VATDEQ＂ and ＂ LGDEQ＂ , TCR BV17 contained ,TDTQ, TCR BVll contained ＂IGQGNTEA＂, and TCR BV20 contained ＂VGLAANEQ＇. Five recipients suffered from pp65 antigenemia in 3 month after transplantation, and pp65-positive cells ranged from 2 to 15 per 5 × 104 white blood cells. Three recipients were CMV-IgM positive. No significant differences were found in TCR BV families between pp65-positive recipients and pp65-negative recipients （ all P 〉 0.05 ）. But there was statistically significant difference in frequency of TCR BV11 between CMV-IgM negative recipients and CMV-IgM positive recipients （P 〈 0. 05）. Conclusion： T cell immune response was characterized by special TCR BV CDR3 spectratyping in HSCT recipients, and TCR BVll expression may be associated with CMV activation.

关 键 词：造血干细胞移植 基因 T细胞受体β/免疫学 互补决定区/免疫学 基因表达谱 巨细胞病毒 聚合酶链反应 

分 类 号：R457.7[医药卫生—治疗学]