正常人外周血和脐血中TCRVβ亚家族sjTRECs的检测情况  

Detection of TCR Vβ subfamily sjTRECs in normal peripheral blood and cord blood

作　　者：尹青松[1] 李扬秋[1] 陈少华[1] 杨力建[1] 卢育洪[1] 张学利[1] 

机构地区：[1]暨南大学医学院血液病研究所,广东广州510632

出　　处：《细胞与分子免疫学杂志》2006年第4期477-479,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30270579);广东省自然科学基金重点项目(23001);广东省教育厅"千百十工程"优秀人才培养基金项目(Q02022);血液病学国务院侨办重点专科基金项目

摘　　要：目的:检测TCR23个Vβ亚家族sjTRECs在正常人外周血中的存在特点,从而进一步了解正常人胸腺近期各Vβ亚家族初始T细胞的输出情况。方法:采用半巢式PCR对4例胸腺、10例脐血和10例正常人外周血分别在2×105、5×104、1×104和1×103个单个核细胞中的23个Vβ亚家族sjTRECs进行扩增。结果:在细胞数相同的情况下,各Vβ-Dβ1sjTRECs的检出率胸腺细胞最高,其次是脐血,正常人最低。当细胞数为2×105、5×104和1×104时,正常人Vβ2、Vβ4、Vβ7、Vβ11和Vβ19亚家族sjTRECs的检出率以及检出sjTRECs的亚家族数量均显著低于脐血。当细胞数为2×105和5×104时,正常人可检测到绝大多数Vβ亚家族的sjTRECs,随着细胞数的降低,部分Vβ亚家族的sjTRECs则检测不到,而少部分Vβ亚家族的sjTRECs则在1000个细胞时仍能检测到。结论:成功建立了半定量检测23个Vβ亚家族sjTRECs的检测方法;在2×105细胞中约可以检测到80%左右的Vβ亚家族初始T细胞,而在5×104细胞中约有半数的Vβ亚家族初始T细胞可检测到,提示不同Vβ亚家族初始T细胞的出现频率有所差异。AIM： To detect the existence of sjTRECs in TCR 23 Vβ subfamilies in normal peripheral blood and cord blood, and to evaluate the recent thymic emigrants of nalve T cells of different TCR Vβ subfamilies. METHODS： Different amounts of DNA from samples （4 cases of thymocytes, 10 cases of cord blood and 10 cases of normal PBMCs） were amplified to estimate the frequency of 23 TCR Vβ-Dβ sjTRECs by using semi-nest PCR. RESULTS： At the same cellular concentration, the most frequency of Vβ-Dβ1 sjTRECs was found in thymocytes, the second was in cord blood, and the lowest was in peripheral blood. At 2 × 10^5, 5 × 10^4 and 1 × 10^4 cells level, sjTRECs of most Vβ subfamilies could be detected in cord blood and peripheral blood, while sjTRECs of some Vβ subfamilies could be detected at 1 × 10^3 cells level. The frequencies of Vβ2-, Vβ4-, Vβ7-, Vβ11- and Vβ19-Dβ1 sjTRECs and the number of detectable Vβ subfamilies from normal PBMCs were obviously lower than those from cord blood. CONCLUSION： The technique of semi-quantitive PCR to detect sjTRECs in 23 Vβ subfamilies was successfully established. Naive T cells from different Vβ subfamilies could be detected with different frequency.

关 键 词：T细胞受体 T细胞受体重排删除环 VΒ亚家族 

分 类 号：R371.23[医药卫生—病原生物学]