摘要
目的:探讨CD154+细胞的表型特征与初始和记忆CD4+ T细胞以及与细胞因子表达之间的关系。方法:正常人外周血单个核细胞(PBMC),经抗CD3+抗CD28单克隆抗体(mAb)刺激培养后,利用多种抗细胞表面分子和抗细胞因子以及抗CD154抗体进行标记,用流式细胞术检测。结果:体外刺激PBMC6h后,CD154表达于CD4+ T细胞,而不表达于CD8+T细胞。对比分析CD4+CD154+ T和CD4+CD154- T细胞上CD45RA(初始)和CD45RO(记忆)表面分子的结果表明,大多数CD4+CD154+ T细胞为记忆细胞。当利用抗CD3或抗CD3+抗CD28mAb刺激后,分泌IFN-γ、IL-2和TNF-α的CD4+ T细胞均为CD4+CD154+ T细胞,而且单个细胞可以同时分泌两种以上细胞因子。进一步研究表明,CD4+CD154+ T细胞低表达或不表达CD25,不表达CD62L,50%左右的细胞表达CCR7。结论:体外短时间刺激PBMC,经过对细胞表型和细胞因子表达的研究,表明CD154分子结合其他表面标记或细胞因子的表达可以用于鉴别初始和记忆CD4+ T细胞。
AIM： To investigate the correlation between CD154^＋ T cells, naive/memory CD4^＋ T cell markers and cytokine expression. METHODS： PBMC from normal persons were isolated, stained by fluorescence-conjugated monoclonal antibodies of surface markers, intracellular cytokines and CD154, and analyzed at the single cell level by flow cytometer. RESULTS： After stimulted in vitro for six hours, CD154 were expressed on CD4^＋ T cells, rather than on CD8^＋ T cells. Analysis of the expression of CD45RA （ marker for naive T ceils） and CD45RO （marker for memory T cells） on CD154^＋ T cells indicated that the majority of CD4^＋ CD154^＋ T cells were memory T cells. When stimulated by antiCD3 or antiCD3 ＋ antiCD28, the cells that produced IFN-γ IL-2 and TNF-α were CD4^＋ CD154^＋ and a single cell produced several cytokines simultaneously. In addition, CD4^＋ CD154^＋ T cells had low level or no expression of CD25 and they didn＇t express CD62L, but approximately fifty percent of the cells expressed CCR7. CONCLUSION： After stimulated in vitro for a short time and through further phenotypic and cytokine analysis, CD154, together with other cell surface markers and cytokines, can be used to discriminate between naive and memory CD4^＋ T cells.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2007年第6期491-494,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家重点基础研究发展规划(973)资助项目(2001CB510007)
广东省自然科学基金团队项目(05200176)
