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Journal of ZheJiang University(Medical Science)  2016, Vol. 45 Issue (5): 515-521    DOI: 10.3785/j.issn.1008-9292.2016.09.10
T cell receptor β-chain CDR3 spectratyping and cytomegalovirus activation in allogeneic hematopoietic stem cell transplant recipients
WU Zhihua1,2, JING Min1, LIANG Hanying1, YANG Rong1, HUANG Yaping1, CHEN Xiaoming1, HU Jianhua1, FAN Jun1
1. State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China;
2. Clinical Laboratory, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China
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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 "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 5×104 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 BV11 expression may be associated with CMV activation.

Key wordsHematopoietic stem cell transplantation      Genes, T-cell receptor beta/immunology      Complementarity determining regions/immunology      Gene expression profiling      Cytomegalovirus      Polymerase chain reaction     
Received: 06 May 2016      Published: 25 September 2016
CLC:  R392  
Cite this article:

WU Zhihua, JING Min, LIANG Hanying, YANG Rong, HUANG Yaping, CHEN Xiaoming, HU Jianhua, FAN Jun. T cell receptor β-chain CDR3 spectratyping and cytomegalovirus activation in allogeneic hematopoietic stem cell transplant recipients. Journal of ZheJiang University(Medical Science), 2016, 45(5): 515-521.

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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TCR BV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TCR BV家族CDR3表达与CMV激活的相关性。结果:3名健康对照者24个TCR BV家族均表达。7例HSCT受者术后TCR BV家族CDR3测序结果显示为BV9、BV11、BV17、BV20等BV家族序列;TCR BV9含“QVRGGTDTQ”,TCR BV11含“VATDEQ”和“LGDEQ”,TCR BV17含“IGQGNTEA”,TCR BV20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/5×104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。TCR BV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TCR BV11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TCR BV家族CDR3谱型,其中TCR BV11的表达可能与CMV激活有关。

关键词: 造血干细胞移植,  基因,  T细胞受体&beta,  /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 
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