用TCRβ可变区基因阵列反映疾病的淋巴细胞克隆变化

Using TCR β variable region gene array in reflecting the alteration of disease lymphocytic clonality.

将正常脐带血TCRβ(T细胞受体 β)重排基因制备成基因阵列 ,观察能否反映肿瘤或自身免疫病患者的淋巴细胞克隆的变化。方法 采用RT -PCR扩增混合 1 0例脐带血的cDNA不同家族的TCRβ可变区基因全部序列 ,经测序胶或SS CP电泳后 ,电转移到尼龙膜上 ,制成TCRβ可变区基因阵列。用同位素α - 32 PdCTP标记正常成人、脐带血、待检患考的TCRβ基因 ,与基因阵列杂交。结果 :从测序胶排列的基基阵列发现 ,正常成人及脐带血呈正常高斯分布 ,即呈多克隆性 ;一些肿瘤患者T淋巴细胞克隆有少数几个克隆的异常增生 ,即呈寡克隆性 ;1例急性T淋巴细胞白血病患者仅在Vβ1 6见单一的条带扩增 ,呈单克隆性。结论 :该基因阵列可以反映正常人淋巴细胞的克隆分布和疾病时出现的淋巴细胞变化。

This experiment use the TCR β rearrangement genes of normal umbilical cord blood mixture to constitute the gene array,and observe if the array reflect the T cell repertoire of tumer and autoimmune diseases.The RNAs are extracted from ten normal full-term fetus umbilical cord blood and mixed,RT-PCR with the different primers of V β gene families are performed,electrophoresis on sequencing gel,and single-strand conformation polymorphism.Then all of these TCR β variable region gene from the gels are transfered to nylon membranes by semi-dry electrophoretic transfer method.The amplified TCR β variable region genes of umbilical cord blood and healthy individuals- and the patients are labeled with α- 32 P dCTP,and hybridized on the arrays.Result:On the gene array of sequencing gel,the healthy adult and normal umbilical cord blood present normal Guassian distribution,means polyclonality.Some patients including four multiple myelomas,three non Hodgki lymphomas show a marked predominance of a few restricted Vβ gene subfamily clone expansion,means oligoclonality.One acute T lymphocyte leukemia patient show a single band of V β 16 family,means monoclonality.Conclusion:The gene array can be used to show lymphocytic clone distribution in normal person and change in patients.