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 Vβsubfamilies in normal peripheral blood and cord blood,and to evaluate the recent thymic emigrants of nalve T cells of different TCR Vβsubfamilies.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-DβsjTRECs 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 Vβsubfamilies could be detected in cord blood and peripheral blood,while sjTRECs of some Vβsubfamilies 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 Vβsubfamilies from normal PBMCs were obviously lower than those from cord blood.CONCLUSION:The technique of semi-quantitive PCR to detect sjTRECs in 23 Vβsubfamilies was successfully established.Naive T cells from different Vβsubfamilies could be detected with different frequency.
作者
尹青松
李扬秋
陈少华
杨力建
卢育洪
张学利
YIN Qing-song;LI Yang-qiu;CHEN Shao-hua;YANG Li-jian;LU Yu-hong;ZHANG Xue-i(Institute of Hematology,Medical College,Jinan University,Guang-zhou 510632,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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
