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TCR Dβ-Jβ sjTRECs检测方法的建立和应用

Establishment and application of the TCR Dβ-Jβ signal joints TRECs detection

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【作者】 李扬秋; 杨力建; 陈少华; 韩素芳; 张学利; 许敏华;

【Author】 LI Yang-qiu, YANG Li-jian, CHEN Shao-hua, HAN Su-fang, ZHANG Xue-li, XU Min-hua (Institute of Hematology, Medical College, Jinan University, Guangzhou 510632, China)

【机构】 暨南大学医学院血液病研究所; 暨南大学医学院血液病研究所 广东广州510632; 广东广州510632; 广东广州510632;

【摘要】 目的 建立检测TCRDβ JβsjTRECs的方法 ,并了解不同T细胞受体Dβ Jβ之间T细胞受体删除DNA环(sjTRECs)在胸腺细胞和外周血T细胞中的形成情况。方法 利用巢式PCR分别扩增 10例正常人外周血单个核细胞和 3例正常胸腺细胞DNA中不同的Dβ片段和Jβ重排时形成的sjTRECs的情况 ,PCR产物进一步进行克隆和序列分析以确定结合区的位置。结果 可检测到Dβ1与 5个Jβ1基因片段、Dβ2与 4个Jβ2基因片段分别形成的sjTRECs,其中以Dβ1 Jβ1S1、Dβ1 Jβ1S2和Dβ2 Jβ2S2的sjTRECs最常见 ,PCR产物经克隆和序列分析证实其形成Dβ JβsjTRECs的情况。 结论 成功地建立了检测Dβ Jβ sjTRECs的方法 ,为分析各TCRβ亚家族的sjTRECs含量和确定TCRβna ve细胞提供了新方法

【Abstract】 Objective To establish the method of Dβ-Jβ sjTRECs detection for analysis the generation of TCR Dβ-Jβ signal joint T-cell receptor excision DNA circles (sjTRECs) within thymocytes and peripheral blood T cells in normal individuals.Methods The different Dβ-Jβ sjTRECs were amplified in genomic DNA from 10 cases of peripheral blood mononuclear cells and 3 cases of thymocytes, by using nested PCR. The PCR products were further cloned and sequenced to define the position of the joints.Results The sjTRECs of Dβ1 with 5 Jβ1 segments and Dβ2 with 4 Jβ2 segments were detected, predominantly in Dβ1-Jβ1S1, Dβ1-Jβ1S2 and Dβ2-Jβ2S2 sjTRECs. The sequences of Dβ-Jβ sjTRECs generation form some PCR products were identified by cloning and sequencing.Conclusions In the present study, the method of detection of Dβ-Jβ sjTRECs was successfully established, it may provide a new technique for analysis of TCRβ subfamily sjTRECs and identify TCRβ nave T cells.

【关键词】 T细胞受体β; sjTRECs; 胸腺细胞; T细胞;
【Key words】 T cell receptor β; sjTRECs; Thymocytes; T cells;
【基金】 国家自然科学基金 (30 2 70 5 79);国家教育部科技重点项目 [教技司 (2 0 0 1) 2 35号 0 10 91];广东省自然科学基金重点项目 (2 30 0 1);广东省教育厅“千百十工程”优秀人才培养基金项目 (Q校 0 2 0 2 2 );广州市科委科技攻关课题 (2 0 0 1 Z 0 37 0 1 3)资助项
  • 【文献出处】 免疫学杂志 ,Immunological Journal , 编辑部邮箱 ,2003年06期
  • 【分类号】R392
  • 【被引频次】12
  • 【下载频次】95
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