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[异基因造血干细胞移植受者的T细胞受体β链互补决定区3谱型分析与巨细胞病毒激活]

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

作者信息

Wu Zhihua, Jing Min, Liang Hanying, Yang Rong, Huang Yaping, Chen Xiaoming, Hu Jianhua, Fan Jun

机构信息

State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China; Clinical Laboratory, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China.

State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.

出版信息

Zhejiang Da Xue Xue Bao Yi Xue Ban. 2016 May 25;45(5):515-521. doi: 10.3785/j.issn.1008-9292.2016.09.10.

Abstract

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). 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. 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 "QVRGGTDTQ", TCR BV11 contained "VATDEQ" and "LGDEQ", TCR BV17 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 5×10 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 >0.05). But there was statistically significant difference in frequency of TCR BV11 between CMV-IgM negative recipients and CMV-IgM positive recipients (<0.05). T cell immune response was characterized by special TCR BV CDR3 spectratyping in HSCT recipients, and TCR BV11 expression may be associated with CMV activation.

摘要

探讨异基因造血干细胞移植(HSCT)受者中T细胞受体β可变区(TCR BV)互补决定区3(CDR3)谱型分析与巨细胞病毒(CMV)激活之间的关联。采用荧光定量PCR熔解曲线分析对7例HSCT受者和3例健康对照者的24个TCR BV家族进行测序。通过免疫组织化学染色检测CMV-pp65抗原血症。采用酶联免疫吸附测定(ELISA)法鉴定针对CMV的血浆IgM。对TCR BV家族与CMV激活之间的关系进行统计学分析。3例健康对照者表达了24个TCR BV家族,而7例受者的TCR BV CDR3测序结果为BV9、BV11、BV17、BV20等。氨基酸序列特征如下:TCR BV9包含“QVRGGTDTQ”,TCR BV11包含“VATDEQ”和“LGDEQ”,TCR BV17包含“IGQGNTEA”,TCR BV20包含“VGLAANEQ”。5例受者在移植后3个月出现pp65抗原血症,每5×10个白细胞中pp65阳性细胞数为2至15个。3例受者CMV-IgM呈阳性。pp65阳性受者与pp65阴性受者之间的TCR BV家族差异无统计学意义(均>0.05)。但CMV-IgM阴性受者与CMV-IgM阳性受者之间TCR BV11的频率差异有统计学意义(<0.05)。HSCT受者的T细胞免疫反应具有特殊的TCR BV CDR3谱型特征,TCR BV11的表达可能与CMV激活有关。

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