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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。
Objective: To analyze the characteristics of TCRVα and Vβ clonal proliferating T cells and CDR3 sequences in peripheral blood of patients with acute promyelocytic leukemia (APL). Methods: TCRVα gene and 24 Vβ gene CDR3 were amplified from peripheral blood mononuclear cells of 1 APL patients by RT-PCR. The positive PCR products were further identified by fluorescent labeling and gene scanning to determine their CDR3 length, and their clonality ; Oligoclonal PCR products were subjected to CDR3 sequence analysis. Results: The oligodendrocyte proliferating T cells in TCRVα6, Vα10 and Vβ23 subfamilies of TCRVα6, Vα10 and Vβ23 subfamily were found in peripheral blood T lymphocyte of APL patients. The sequences of TCRVα6, Vα10 and Vβ23 subfamily genes were Vα6NJα17, Vα10NJα35 and Vβ23NDβ1NJβ2.7 The amino acid sequences of CDR3 were CAMRENSAGNK, YLCAGDELLWECA and CASSSKWAG-GTYEQY, respectively. The three TCR genes were deposited with GenBank (accession number: EU544946, EU544947 and EU647219). CONCLUSION: TCRVα6, Vα10 and Vβ23 clonally proliferating T cells in peripheral blood T cells of patients with APL may be antigen-specific immune responses caused by the body against leukemia cells. These three unique TCR rearrangements will further serve as a basis for carrying out APL-specific Immunotherapy research to provide information.