【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/5×10~4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。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。更多还原

【Abstract】 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 x 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 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.更多还原