【摘要】 目的:分析急性早幼粒细胞白血病(APL)患者外周血TCRVα和Vβ克隆性增殖T细胞及CDR3序列特点。方法:利用RT-PCR法扩增1例APL患者外周血单个核细胞中29个TCRVα基因和24个Vβ基因CDR3,阳性PCR产物进一步经荧光标记和基因扫描分析确定其CDR3长度,了解其克隆性;寡克隆的PCR产物再进行CDR3区序列分析。结果:该例APL患者外周血T细胞分别在TCRVα6、Vα10和Vβ23亚家族中出现寡克隆性增殖T细胞,经序列分析显示TCRVα6、Vα10和Vβ23亚家族基因序列分别为Vα6NJα17、Vα10NJα35和Vβ23NDβ1NJβ2.7,其CDR3区氨基酸序列分别为CAMRENSAGNK,YLCAGDELLWECA,CASSSKWAG-GTYEQY,该3个TCR基因已被GenBank收录(登陆号:EU544946、EU544947和EU647219)。结论:APL患者外周血T细胞出现的TCRVα6、Vα10和Vβ23克隆性增殖T细胞可能是机体对抗白血病细胞所引起的抗原特异性免疫反应;这3个独特TCR重排序列将进一步作为开展APL特异性免疫治疗研究提供资料。更多还原

【Abstract】 AIM:To investigate the clonality and the pattern of CDR3 sequence of TCR Vα and Vβ repertoire in patients with acute promyelocytic leukemia(APL).METHODS:The CDR3 of TCR Vα 29 and Vβ 24 subfamily genes were amplified in mononuclear cells from peripheral blood of one case with APL using RT-PCR.The positive PCR products were further labeled with fluorescent and analyzed by genescan technique for the CDR3 size,to evaluate clonality of the detectable TCR Vα and Vβ T cells.The products of the oligoclonal T cells were analyzed by sequencing to define the sequence of CDR3.RESULTS:Oligoclonal T cells expressed TCR Vα6,Vα10 or Vβ23 genes were identified from the APL patient.The molecular characteristics of the CDR3 of Vα and Vβ genes rearrangement were Vα6NJα17,Vα10NJα35 and Vβ23NDβ1NJβ2.7.The amino acid sequence of CDR3 region in TCR Vα 6,Vα10 and Vβ23 subfamilies were CAMRENSAGNK,YLCAGDELLWECA or CASSSKWAGGTYEQY,respectively.These sequences were accepted by GenBank(Accession Number EU544946,EU544947 and EU647219).CONCLUSION:Three idiotype Vα and Vβ sequeces are identified in a case with APL,which is thought that may mediate the host specific immunity response for leukemia cells in APL.It might provide data for farther research on the specific immunotherapy of APL.更多还原