Yang Li-Jian, Li Yang-Qiu, Chen Shao-Hua, Han Su-Fang, Chen Sheng-Ting, Zhang Xue-Li, Zhang Tao
Institute of Hematology, Jinan University Medical College, Guangzhou 510632, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2003 Oct;11(5):499-502.
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.
为分析急性早幼粒细胞白血病（APL）患者体内及T细胞培养后体外TCR Vβ亚家族T细胞的分布及克隆性扩增，采用液体T淋巴细胞培养技术，用重组人白细胞介素-2（rhIL-2）和抗CD3抗体对3例APL患者的外周血单个核细胞进行扩增。采用逆转录聚合酶链反应（RT-PCR）对3例APL患者T细胞培养前后TCRβ可变区基因的互补决定区3（CDR3）进行扩增。对阳性产物进一步分析，通过基因扫描鉴定T细胞的克隆性。结果显示，患者T细胞中仅检测到24个Vβ亚家族中的一部分，T细胞培养后可鉴定出一些Vβ亚家族T细胞。所有患者均能发现部分TCR Vβ亚家族中的克隆性扩增T细胞。在T细胞培养后的不同时间点，两例患者检测到Vβ1、Vβ3、Vβ7、Vβ16和Vβ20 T细胞有相似的寡克隆扩增。结论：患者T细胞中TCR Vβ亚家族的限制性表达可能是白血病的共同特征。体外T细胞培养后可诱导出一些Vβ亚家族T细胞。T细胞培养后不同时间点TCR Vβ亚家族T细胞的持续克隆性扩增可能是患者T细胞针对特定APL细胞相关抗原的特异性免疫反应。
