急性早幼粒细胞白血病患者中TCR Vβ T细胞体内外克隆性增殖的比较

为了解急性早幼粒细胞白血病(APL)患者T细胞在体内或体外经诱导后TCR Vβ亚家族的分布和克隆性增殖特点。利用T淋巴细胞液体培养法。在rhIL—2和抗CD3单抗条件下诱导扩增APL患者单个核细胞，并应用RT-PCR分别扩增培养前后患者T细胞的TCR Vβ24个亚家族基因的互补决定区3(CDR3)片段。了解各Vβ亚家族的表达情况；对阳性的PCR产物进一步经荧光素标记和基因扫描分析产物的CDR3长度。了解T细胞克隆性。结果发现，APL患者T细胞仅表达部分Vβ亚家族。但经体外培养后可检测到部分新增TCR Vβ亚家族T细胞。全部患者存在某些。TCR Vβ亚家族的克隆性增殖T细胞。2例患者均出现相似的Vβ1，Vβ3，Vβ7，Vβ16和Vβ20 T细胞的克隆性增殖情况。结论：T细胞的体外培养可诱导某些Vβ亚家族的表达。在T细胞培养不同时间中，呈持续克隆性增殖的TCR Vβ亚家族T细胞可能是患者T细胞对APL白血病细胞相关抗原的特异性免疲应答。

Comparison of the Clonal Expansion of TCR Vβ T Cells in Patients with Acute Promyelocytic Leukemia In Vivo and In Vitro

In order to analyze the distribution and clonal expansion of TCR Vbeta subfamily T cells in patients with acute promyelocytic leukemia (APL) in vivo and in vitro after T cell culture, the peripheral blood mononuclear cells from 3 APL patients were expanded by rhIL-2 and anti-CD3 antibody using liquid T lymphocytes culture technique. The complementary determining region 3 (CDR3) of TCR beta with variable region genes was amplified in T cells from 3 APL cases before and after T cell culture by using RT-PCR. The positive products were further analyzed to identify the clonality of T cells by genescan. The results showed that only a part of 24 Vbeta subfamilies was detected in T cells from the patients, and some Vbeta subfamily T cells could be identified after T cells culture. The clonal expansion T cells in some TCR Vbeta subfamilies could be found in all patients. The similar oligoclonal expansion of Vbeta1, Vbeta3, Vbeta7, Vbeta16 and Vbeta20 T cells was detected in two cases at different time points after T cell culture. It is concluded that the restricted expression of TCR Vbeta subfamily in T cells from patients might be the common feature in leukemia. Some Vbeta subfamily T cells could be induced after T cells culture in vitro. The continual clonal expansion of TCR Vbeta subfamily T cells at different time points after T cells culture could be a specific immune response of patients T cells related to the specific APL cell associated antigen.