Zhu P, Li J, Ma M
Department of Hematology, The First Teaching Hospital, Beijing University, Beijing 100034, China.
Zhonghua Yi Xue Za Zhi. 2000 Sep;80(9):668-71.
To analyze T lymphocytic clone formation in patients with malignant disease after bone marrow or peripheral blood stem cell transplantation and to understand the relationship between immunological reconstitution and clinic remission.
Due to the different TCRgamma gene rearrangements in different gamma/delta T cell clones, a gene fingerprinting was set up by PCR and denatured gel electrophoresis. The gene fingerprinting was used to analyze the gamma-delta T cell clone variations in 8 patients who received stem transplantation. In order to identify the source of the regenerated lymphocytes, the polymorphism of microsatellite was used at mean time.
After PCR amplification and electrophoresis of each TCRgammaV region gene on a 60 cm sequence gel, a gene fingerprinting composed of about a hundred bands was set up. It showed the basic distribution of the dominant gamma/delta T cell clones in the body. Usually, a new band presenting after transplantation represented an expansive lymphocytic clone. Successful immunological reconstitution led to a good prognosis; and unsuccessful immunological reconstitution led to bad prognosis. After transplantation, some T cell clones expanded. Comparing the sequences of a expansive T cell clone in a multiple myeloma patient after auto-stem cell transplantation with 230 000 genes of immunoglobulin and T cell receptor in GeneBank, we found that two T cell line had homogenous sequences. Both T cell clones recognized homogeneous antigen.
A new method to observe the distribution of gamma/delta T cells called gene fingerprinting was set up. Gene fingerprinting is useful in the analysis of immunological reconstitution after stem cells transplantation. The effect of transplantation is correlated to immunological reconstitution. Transplantation patients may appear some expansive T cell clones; probably they have anti - tumor effect.
分析恶性疾病患者骨髓或外周血干细胞移植后T淋巴细胞克隆的形成情况,了解免疫重建与临床缓解之间的关系。
由于不同的γ/δT细胞克隆中TCRγ基因重排不同,通过聚合酶链反应(PCR)和变性凝胶电泳建立基因指纹图谱。利用该基因指纹图谱分析8例接受干细胞移植患者的γδT细胞克隆变化。同时,为了确定再生淋巴细胞的来源,采用微卫星多态性分析。
在60cm序列胶上对每个TCRγV区基因进行PCR扩增和电泳后,建立了由约100条带组成的基因指纹图谱。它显示了体内优势γ/δT细胞克隆的基本分布。通常,移植后出现的新条带代表一个扩增的淋巴细胞克隆。成功的免疫重建导致良好的预后;而免疫重建失败则导致不良预后。移植后,一些T细胞克隆扩增。将一名多发性骨髓瘤患者自体干细胞移植后一个扩增的T细胞克隆序列与基因库中23万个免疫球蛋白和T细胞受体基因进行比较,发现两个T细胞系具有同源序列。两个T细胞克隆识别同源抗原。
建立了一种观察γ/δT细胞分布的新方法——基因指纹图谱。基因指纹图谱有助于分析干细胞移植后的免疫重建情况。移植效果与免疫重建相关。移植患者可能会出现一些扩增的T细胞克隆;它们可能具有抗肿瘤作用。