用RT-PCR和Genescan分析CML急变期病人TCRVβT细胞的表达和克隆性

Analysis of the Expression of TCR Vβ Subfamily Genes and T Cells Clonality in Blast Crisis of CML Using RT-PCR and Cenescan

目的：了解CML急变期的TCRVβ亚家族T细胞的表达及其克隆性。方法：采用RT－PCR扩增4例CML急变期病人的外周血单个核细胞的TCRVβ24个亚家族的互补决定区3（CDR3），产物进一步经基因扫描分析确定T细胞的克隆性。结果：病人外周血仅表达4～9个Vβ亚家族T细胞，主要为Vβ1，Vβ2，Vβ3，Vβ5，Vβ15和Vβ17；基因扫描分析显示2例CML急粒变的部分产物为寡克隆性，而2例CML急淋变者的产物均为多克隆性。结论：CML急变期外周血仅选择性表达部分Vβ亚家族T细胞；CML急粒变存在克隆性增殖T细胞，这可能是机体对白血病细胞相关抗原的一种直接反应。该方法可用于检测微小残留病变和判断疾病复发。

objective The distribution of TCR V6 subfamily T cells and clonality were analyzed in blast crisis of chronic myeloid leukemia(CML). Methods The complementarity determining region 3(CDR3) of TCRVβ 24 subfamily genes in peripheral blood mononuclear cells form 4 cases with blast crisis of CML was amplified us ing RT-PCR,the products were further by Genescan analysis to identify the clonality of T cells. Results Only 4~9 Vβ subfamily T cells were found in samples from blast phase of CML, the Genescan analysis showed that individual Vβ subfamily products from two myeloid blast crisis of CML samples exhibited oligoclonality, whereas all products from another two cases with lymphoid blast crisis of CML were polyclonality. Conclusion TCR Vβ subfamily T cells selective express on blast crisis of CML, T cell clonality expansion may be found in myeloid blast crisis of CML cases,suggesting that may indicate a host response directed against leukemia cell relatION antigen. This analysls method may be used for detection of minimal residual disease and the relapse of disease.