摘要
目的:以佛波醇酯加离子霉素作为刺激剂,验证CD4+CD25+调节性T细胞本身并不存在分泌IL-2障碍;同时通过对脐血和成人外周血的比较性研究,了解脐血CD4+CD25+T细胞的成熟度。方法:以autoMACS从足月婴儿脐血(CB)和成人外周血(PB)分选CD4+CD25+和CD4+CD25-T细胞,以PDB+ionomycin作为刺激剂,培养45h后流式细胞术检测各组细胞表达CD69和CD25水平,并以Luminex多重细胞因子检测技术检测培养上清中7种细胞因子的浓度。结果:经PDB+ionomycin刺激后,CB、PB的CD4+CD25+和CD4+CD25-T细胞均发生增殖,但在培养45h后CD4+CD25+T细胞均出现细胞状态变差或死亡倾向。CB、PB的CD4+CD25+T细胞活化后CD25分子表达进一步上调,高于CD25-细胞活化后的CD25分子密度。经PDB+ionomycin刺激后,PB CD4+CD25+和CD4+CD25-T细胞均分泌高水平的IFN-γ、IL-2和TNF-α,但CD25+细胞分泌IL-5、IL-4和IL-10水平远远高于CD25-细胞;CBCD4+CD25+和CD4+CD25-T细胞亦分泌高水平的IL-2和TNF-α,但IFN-γ水平远远低于PB,基本不分泌IL-5、IL-4和IL-10。结论:CD4+CD25+T细胞本身并不存在合成和分泌IL-2障碍,其可能具有与传统T细胞不同的T细胞受体信息转导模式;脐血CD4+CD25+T细胞功能尚未完全成熟。
AIM： To confirm that CD4^＋CD25^＋ regulatory T cells don＇t have an instinctive defection in IL-2 secretion, and to have an insight into the maturation state of CD4^＋CD25^＋T cells in cord blood. METHODS： CD4^＋CD25^＋ and CD4^＋CD25^-T cells were purified from cord blood of term infants （CB） and adult peripheral blood （PB） by antoMACS, and stimulated with PDB plus ionomycin. After 45 hours of culture, cells were detected for expression of CD69 and CD25 by flow cytometry, and the supematants were measured for 7 kinds of cytokines by Luminex. RESULTS：CD4^＋CD25^＋T cells fnan both CB and PB proliferated comparably with CD4^＋CD25^-T cells when stimulated with PDB plus ionomycin. After 45 hours of culture, however, the CD4^＋CD25^＋T cells underwent a tendency of cell death. Expression of CD25 was further upregulated when CD25^＋ cells were activated. Under stimulation of PDB plus ionomycin, both CD4^＋CD25^＋ and CD4^＋CD25^＋T cells in PB secreted high levels of IFN - γ, IL- 2 and TNF-α, with CD25^＋ cells secreted much higher level of IL- 5, IL- 4 and IL- 10 than those in CD25^- cells;CD4^＋CD25^＋ and CD4^＋CD25^＋T cells in CB also secreted high level of IL- 2 and TNF -α but much lower level of IFN - γ than those in PB, and no secretion of IL- 5, IL- 4 and IL- 10 was observed. CONCLUSION： CD4^＋CD25^＋ regulatory T cells don＇ t have an instinctive defection in IL-2 secretion, otherwise there may be a different TCR signaling pattern in CD4^＋CD25^＋T cells from traditional T cells. The CD4^＋CD25^＋Tcells in cord blood have not fully matured in function.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第6期1133-1137,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金重点项目(No.30230305)
国家自然科学基金面上项目(No.30500466)
广东省自然科学基金博士启动项目(No.05300419)
