基因扫描分析CML病人外周血的T细胞克隆性

 应用RT-PCR方法扩增12例CML病人外周血单个核细胞的T细胞受体(CR)Vβ24个亚家族基因的互补决定区3(CDR3), 了解VβT细胞的分布情况, 8例正常人和T细胞株K37作为对照。 PCR产物进一步经基因扫描分析确定T细胞克隆性。 结果发现12例CML外周血中分别表达3－21Vβ亚家族T细胞, 8例正常人则表达除Vβ20外的全部VβT细胞, K37细胞仅表达Vβ9。基因扫描分析显示12例CML中8例病人的部分Vβ亚家族PCR产物至一主峰图象, 提示为克隆性生长的T细胞。 推测这是宿主对白血病细胞相关抗原的一种直接反应。 该方法有可能作为白血病特异性免疫治疗的临床研究的一种新的分子生物学研究方法。

Identification of Peripheral blood T cell clonality in CML by genescan analysis

The distribution of TCR Vβsubfamilies T cells was analyzed using RT - PCR,which amplified the Complementarity determining region 3(CDR3)of TCR Vβgene inperipheral blood mononuclear cell(PBMC) with chronic myeloid leukemia(CML), 8cases normal samples and T cell line K37 serve as control. The PCR Products were further studied by genescan analysis to identify tO the T cell clonality. The expression of 3-21Vβ subfamilies was found in PBMC from 12CML cases, besides Vβ20, all subfamilyT cells of TCR Vβwere identified in PBMC of 8 control cases, but only one Vβsubfamily(Vβ)was expressed in T cell line K37. The genescan analysis showed that some Vβsubfamily products from 8 of 12 CML cases contained a dominant peak. The result indicatedthat T cell clonality expansion may be found in CML cases. Suggesting that may indicatea host response directed against leukemia cell related antigen. This analysis method maybe useful as a new molecular biologic analysis tool for clinical study of specific immunetherapy in leukemia.