摘要
本研究旨在观察CD62L-/CD62L+T细胞应答抗原的能力,了解CD62L分子在移植物抗宿主病(GVHD)中的作用。用DC细胞呈递特异性抗原hCD4刺激T细胞,流式细胞术检测其表面CD62L分子表达;对分选获得的CD62L-/CD62L+两群T细胞再次给予新的抗原(EL4细胞冻融抗原)刺激,用流式细胞术检测CD62L-及CD62L+T细胞在前后两次受到不同抗原刺激后的活化增殖能力。结果显示:T细胞表面CD62L+分子在受到抗原刺激后表达下调,CD62L+细胞由69.34%降至36.75%,相应的CD62L-T细胞由30.04%升至63.15%。比较第1天及第7天在高CFSE荧光值区(M1)的细胞数表明,在特异性抗原(hCD4)的刺激下,CD62L+T细胞平均百分比值分别为(87.88±1.08)%,(86.88±1.46)%(p>0.05);CD62L-T的测得值分别为(84.52±2.73)%(84.71±2.33)%(p>0.05)。两群细胞数在第7天与第1天相比均未出现明显增殖;在非特异性抗原刺激下,CD62L+T细胞在第1天和第6天于M1区的细胞数平均百分比值分别为(76.49±2.41)%,(22.25±3.29)(p<0.001);CD62L-T测得值分别为(77.35±3.82)%,(74.76±3.90)%(p>0,05)。此结果说明CD62L+T细胞发生了分裂增殖,而CD62L-T细胞群则没有出现类似的情况。结论:通过体外方法获得了CD62L-CD62L+T细胞,这两类细胞在特异性抗原刺激下未发生增殖反应,但在非特异性抗原刺激下CD62L+T细胞发生了分裂增殖,而CD62L-T细胞无此反应。此结果表明CD62L+T细胞在GVHD发生中有着重要作用,同时也为临床上通过体外方法获得CD62L-T细胞而选择性地去除移植物中CD62L+T细胞以减少GVHD的发生提供了思路。
The aim of this study was to investigate the ability of CD62L^-/CD62L^＋ T cells responding to antigens so as to explore the role of CD62L in GVHD. The T cells were stimulated by DCs-presenting specific antigen hCD4; the expression of CD62L on surface of T cells was detected by flow cytometry, the CD62^- T/CD62L^＋ T cells were gained by sorting selection method, then were stimulated with a new antigen （freeze-thaw antigen of EL4 cells）, and the proliferation activity of CD62L^- and CD62L^＋ T cells was detected by flow cytometry after stimulating with different antigens at twice. The results showed that after stimulating with antigen, the expression level of CD62L^＋ on surface of T cells was down-regulated from 60.34% to 36.75%, and the expression of CD62L^- on surface of T cells increased from 30.04% to 63.15%. Compared with cells counts in area with high CFSE （M1）, the average percentages of CD62L^＋ T cells under stimulating with specific antigen （hCD4） on days 1 and 6 were （ 87.88±1.08 ） % and （ 86.88±1.46 ） % respectively （p 〉 0.05 ）, and the average percentages of CD62L^＋ T cells after stimulating with specific antigen on days 1 and 6 were （84.52±2.73）% and （84.21±2.33）% respectively （p 〉0.05）, there was no significant difference between two T cell groups on days 1 and 6. After stimulating with non-specific antigen, the average percentages of CD62L^＋ T cells in M1 area on days 1 and6 were （76.49±2.41）% and （22.25±3.29）% respectively （p 〈0.001）, the average percentages of CD621^-T cells under conditions mentioned above were （77.35±3.82）% and （74.76±3.90）% respectively （p 〈 0.05 ）. The results indicated that the proliferation activity of CD62L^＋ T cells appeared, and the proliferation of CD62L^＋ T cells did not occur. It is concluded that the CD62L^- T/CD621^＋ T cells are gained in vitro, no proliferation reaction occurred in these two kinds of cells under stimulation of specific antigen, but the mitotic proliferation
出处
《中国实验血液学杂志》
CAS
CSCD
2009年第1期170-175,共6页
Journal of Experimental Hematology
基金
国家自然科学基金资助项目
编号30671980
