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| 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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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 5×104 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.
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Received: 06 May 2016
Published: 25 September 2016
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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
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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| [[1]] |
SERVAIS S, LENGLINE E, PORCHER R, et al.Long term immune reconstitution and infection burden after mismatched hematopoietic stem cell transplantation[J].Biol Blood Marrow Transplant, 2014, 20(4):507-517.
|
|
|
| [[2]] |
KHARFAN-DABAJA M A, BOECKH M, WILCK M B, et al.A novel therapeutic cytomegalovirus DNA vaccine in allogeneic haemopoietic stem-cell transplantation:a randomised, double-blind, placebo-controlled, phase 2 trial[J].Lancet Infect Dis, 2012, 12(4):290-299.
|
|
|
| [[3]] |
MOINS-TEISSERENC H, BUSSON M, SCIEUX C, et al.Patterns of cytomegalovirus reactivation are associated with distinct evolutive profiles of immune reconstitution after allogeneic hematopoietic stem cell transplantation[J].J Infect Dis, 2008, 198(6):818-826.
|
|
|
| [[4]] |
BOECKH M, BOWDEN R A, GOOLEY T, et al.Successful modification of a pp65 antigenemia-based early treatment strategy for prevention of cytomegalovirus disease in allogeneic marrow transplant recipients[J].Blood, 1999, 93(5):1781-1782.
|
|
|
| [[5]] |
闫淑媛, 陈平洋.人巨细胞病毒感染的实验室诊断研究进展[J].国外医学:儿科学分册, 2005, 32(5):284-287.YAN Shuyuan, CHEN Pingyang.The development of laboratory diagnosis of human cytomegalovirus infection in neonates[J].Foreign Medical Sciences Section of Pediatric, 2005, 32(5):284-287.(in Chinese)
|
|
|
| [[6]] |
杨介钻, 李兰娟.TCR β链CDR3概况与感染性疾病的研究[J].国际流行病学传染病学杂志, 2009, 36(2):39-42.YANG Jiezuan, LI Lanjuan.Study for complementarity determining region 3 of T cell receptor beta chain in infectious diseases[J].International Journal of Epidemiology and Infectious Disease, 2009, 36(2):39-42.(in Chinese)
|
|
|
| [[7]] |
RUGGIERO E, NICOLAY J P, FRONZA R, et al.High-resolution analysis of the human T-cell receptor repertoire[J].Nat Commun, 2015, 6:8081.
|
|
|
| [[8]] |
NAKASONE H, TANAKA Y, YAMAZAKI R, et al.Single-cell T-cell receptor-β analysis of HLA-A*2402-restricted CMV-pp65-specific cytotoxic T-cells in allogeneic hematopoietic SCT[J].Bone Marrow Transplantation, 2014, 49(1):87-94.
|
|
|
| [[9]] |
罗荣, 马锐, 孙万邦, 等."荧光定量PCR溶解曲线法"监测人TCRβ链CDR3谱系漂移技术的建立[J].免疫学杂志, 2009, 25(4):446-451.LUO Rong, MA Rui, SUN Wanbang, et al.Establishment of the real-time fluorescence quantitative reverse transcription polumerase chain reaction with DNA melting curve analysis for detecting the CDR3 skewing of TCR beta gene repertoire in the human peripheral blood[J].Immunological Journal, 2009, 25(4):446-451.(in Chinese)
|
|
|
| [[10]] |
BUNDE T, KIRCHNER A, HOFFMEISTER B, et al.Protection from cytomegalovirus after transplantation is correlated with immediate early 1-specific CD8 T cells[J].J Exp Med, 2005, 201(7):1031-1036.
|
|
|
| [[11]] |
GUERRERO A, RIDDELL S R, STOREK J, et al.Cytomegalovirus viral load and virus-specific immune reconstitution after peripheral blood stem cell versus bone marrow transplantation[J].Biol Blood Marrow Transplant, 2012, 18(1):66-75.
|
|
|
| [[12]] |
LIU A B, MA Y D, FAN J, et al.Evaluation of human cytomegalovirus-specific CD8+T-cells in allogeneic haematopoietic stem cell transplant recipients using pentamer and interferon-enzyme-linked immunospot assays[J].J Clin Virol, 2013, 58(2):427-431.
|
|
|
| [[13]] |
SCHMIDT-HIEBER M, LABOPIN M, BEELEN D, et al.CMV serostatus still has an important prognostic impact in de novo acute leukemia patients after allogeneic stem cell transplantation:a report from the Acute Leukemia Working Party of EBMT[J].Blood, 2013, 122(19):3359-3364.
|
|
|
| [[14]] |
WANG G C, DASH P, MCCULLERS J A, et al.T-cell receptor αβ diversity inversely correlates with pathogen-specific antibody levels in human cytomegalovirus infection[J].Sci Transl Med, 2012, 4(128):128ra4.
|
|
|
| [[15]] |
ENGSTRAND M, LIDEHALL A K, TOTTERMAN T H, et al.Cellular responses to cytomegalovirus in immunosuppressed patients:circulating CD8+T cells recognizing CMVpp65 are present but display functional impairment[J].Clin Exp Immunol, 2003, 132(1):96-104.
|
|
|
| [[16]] |
OZDEMIR E, ST JOHN L S, GILLESPIE G, et al.Cytomegalovirus reactivation following allogeneic stem cell transplantation is associated with the presence of dysfunctional antigen-specific CD8+T cells[J].Blood, 2002, 100(10):3690-3697.
|
|
|
| [[17]] |
王前明, 宋秀宇, 洪强, 等.慢性乙型肝炎病毒感染者辅助性T淋巴细胞亚群的变化[J].检验医学与临床, 2008, 5(12):707-707.WANG Qianming, SONG Xiuyu, HONG Qiang, et al.Variance of helper lymphocyte T cell subsets in patients with chronic hepatitis B virus infection[J].Journal of Laboratory Medicine and Clinical Sciences, 2008, 5(12):707-707.(in Chinese)
|
|
|
| [[18]] |
YANG J, HE J, HUANG H, et al.Molecular characterization of T cell receptor beta variable in the peripheral blood T cell repertoire in subjects with active tuberculosis or latent tuberculosis infection[J].BMC Infect Dis, 2013, 13:423.
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