论文部分内容阅读
利用巢式或半巢式PCR扩增10例正常人外周血单个核细胞(PBMC)、4例分选CD3~+细胞和7例正常胸腺细胞DNA中TCR δRec区与Jδ1、Dδ3和Ja重排的基因片段,分析正常人外周血T细胞和胸腺细胞中TCR δRec基因重排情况。克隆性PCR产物进一步进行核苷酸序列分析确定其重排位置。结果发现了4种新的TCR δRec重排,包括TCR δRec_(149321)-Jδ1、TCRδRec_(149820)-Jδ1、TCR δRec_(151657)(Nx)-Ja和TCR δRec_(153199)-Jδ1等,其中以TCR δNx的重排最多见,通过利用不同模板DNA的PCR分析发现δRec重排在外周血和胸腺细胞中有所不同。结果显示TCR δNx-Jδ1重排在成熟和不成熟T细胞发生频率均较高,而TCR δNx-Dδ3重排在不成熟T细胞中发生率较高。但所有重排均不表达于mRNA中。本研究结果为TCR δ基因重排的研究补充了一些新的数据。
The TCR δRec region of DNA from 10 normal human peripheral blood mononuclear cells (PBMCs), 4 sorted CD3 ~ + cells and 7 normal thymocytes was amplified by nested or semi-nested PCR with Jδ1, Dδ3 and Ja rearrangements The gene fragment of TCR δRec gene in peripheral blood T cells and thymocytes was analyzed. The cloned PCR product was further subjected to nucleotide sequence analysis to confirm its rearrangement position. As a result, four new TCR δRec rearrangements were found, including TCR δRec_ (149321) -Jδ1, TCRδRec_ (149820) -Jδ1, TCR δRec_ (151657) (Nx) -Ja and TCR δRec_ (153199) -Jδ1, Rearrangement of TCR δNx was the most common, and δRec rearrangement was found to be different in peripheral blood and thymocytes by PCR analysis using different template DNA. The results showed that TCR δNx-Jδ1 rearrangements had a higher frequency of mature and immature T cells, while TCR δNx-Dδ3 rearrangements had a higher incidence in immature T cells. However, all rearrangements were not expressed in mRNA. Our results add some new data for the study of TCR δ gene rearrangement.