摘要
目的观察哮喘小鼠脾脏CD4+CD25+调节性T细胞(简称Treg细胞)数量及功能的改变。方法以屋尘螨提取液复制小鼠哮喘模型后,分离小鼠脾脏CD4+T细胞,采用流式细胞仪检测Treg细胞占CD4+T细胞的比例;分离Treg细胞,与屋尘螨提取液共同培养后测定上清液中白细胞介素-4(IL-4)及IL-10的含量;将Treg细胞与CD4+T细胞共同培养,观察其对CD4+T细胞增殖及IL-4、IL-5、γ-干扰素(IFN-γ)分泌的影响。结果哮喘小鼠脾脏Treg细胞占CD4+T细胞比例明显下降;与正常组相比,哮喘小鼠脾脏Treg细胞IL-4、IL-10的分泌无明显差异;Treg细胞可显著抑制哮喘小鼠CD4+T细胞的增殖,并降低其IL-4的分泌,对IFN-γ的合成分泌无明显作用;使用anti-CD25 mAb后,哮喘和正常小鼠Treg细胞的作用均被显著抑制。结论哮喘小鼠Treg细胞功能与正常对照组小鼠无明显区别,但数量显著下降。哮喘发病过程中Th2型反应占优势,与Treg细胞数量减少,对Th2型反应的抑制作用降低有一定关系。
Objective To observe the change of CD^4＋ CD25^＋ regulatory T cells （Treg cells） and its function in mouse model of asthma. Methods C57BL/6J mice were sensitized and challenged with standardized house dust mite extract to established asthma model, The spleen cells were harvested and spleen dendritic cells （DCs） and CD^4＋T cells were isolated. The percentage of Treg cells in CD^4＋T cells was calculated with flow cytometry and Treg cells were isolated from CD^4＋T cells.Then Treg cells were cultured with house dust mite extract（HDME） and the supernatants were collected for cytokine IL-4 and IL-10 assay. After co-incubated with Treg cells, DCs and HDME, with or without anti-CD25 mAb, the CD^4＋T cells were harvested for the measurement of expansion with MTT while the supernatants were collected for IL-4, IL-10 and IFN-γassay. Results The number and proportion of Treg cells in spleen were decreased in the asthmatic group, however, the ability of Treg cells to secret IL-4 and IL-10 was not different between asthmatic and control animals.The cell proliferation and IL-4 secretion of CD^4＋T cells were suppressd by Treg cells in asthmatic mouse but secretion of IL-10 and IFN-γ could not be inhibited. The effects of Treg cells on CD^4＋T cells were abolished by anit-CD25 mAb both in asthmatic and in control animals. Conclusions Asthma animals have Treg cells of less amount and normal function which may contribute to the up-regulated Th2 response in asthma.
出处
《中国呼吸与危重监护杂志》
CAS
2005年第6期455-458,共4页
Chinese Journal of Respiratory and Critical Care Medicine
基金
国家自然科学基金资助(30470772)
