摘要
目的 探讨腹腔注射高迁移率族蛋白B1（HMGB1）后不同基因型小鼠天然调节性T细胞（CD4^＋CD25^＋Treg）免疫功能的改变及其受体作用机制.方法 分别给C3H/HeN和C3H/HeJ[分别为Toll样受体4（TLR4）野生型（TLR4^＋/＋）和天然突变型（TLR4^-/-]小鼠腹腔注射不同剂量HMGB1（0、10、20μg/只）,饲养48 h后采用免疫磁珠法分离小鼠脾脏CD4^＋CD25^＋Treg.体外培养12 h后采用流式细胞仪检测CD4^＋CD25^＋Treg表达细胞毒性T淋巴细胞相关抗原-4（CTLA-4）表达强度,并应用酶联免疫吸附试验（ELISA）检测CD4^＋CD25^＋Treg生成白细胞介素（IL）-10量.结果 与对照组比较,20 μg HMGB1攻击后C3H/HeN小鼠CD4^＋CD25^＋Treg表达CTLA-4水平显著下降（78.70±11.42与60.76±7.64,P〈0.01）,同时细胞牛成IL-10量也明显降低[（96.89±6.25）ng/L与（47.11±4.25）ng/L,P〈0.01];但不同剂量HMGB1攻击可引起C3H/HeJ小鼠CD4^＋CD25^＋Treg表达CTLA-4明显上调和生成IL-10量不同程度地增加（P〈0.01）.结论 HMGB1攻击可显著影响CD4^＋CD25^＋Treg介导的免疫状态,TLR4在HMGB1诱导CD4^＋CD25^＋Treg免疫活性过程中发挥了重要负向调控作用.
Objective To investigate the effect of high mobility group box-1 protein（HMGB1） on immune function of regulatory T cells（Tregs） in two gene-type mice and its potential receptor mechanism in vivo.Methods 10 μg or 20 μg HMGB1 was administered i.P.in C3H/HeN（TLR4 wild type,TLR4^＋/＋）and C3H/HeJ （TLR4 mutant type,TLR4^-/-）mice.Mice were sacrificed at the 48th h after HMGB1 challenge,and splenic CD4^＋CD25^＋ Tregs were harvested by magnetic beads aseptically following another 12 h culture.Cytotoxic T lymphocyte-associated antigen 4（CTLA-4）expressed on CD4^＋CD25^＋＋ Tregs was detected by means of flow cytometry,and the production of interleukin（IL）-10 in supematants was examined by ELISA.Results The expression levels of CTLA-4 in two gene-type mice were similar （P〉0.05）.Compared to the controls,the expression of CTLA-4 on CD4^＋CD25^＋＋ Tregs was markedly down-regulated in C3H/HeN mice with a maximal response at a dose of 20μg HMGB1（78.70±1 1.42 vs 60.76±7.64,P〈0.01）,and IL-10 release was also significantly decreased[（96.89±6.25）vs（47.11±4.25）ng/L,P〈0.01].However,after treatment with various doses of HMGB1 in C3H/HeJ mice.the expression of CTLA-4 on CD4^＋CD25^＋＋ Tregs and IL-10 release were markedly enhanced compared to the controls（P〈0.01）.Conclusion HMGB1 challenge can markedly influence the inununological activity of CD4^＋CD25^＋＋ Tregs in mice,and TLR4 appears to be critically involved in down-regulation of HMGB1-mediated CTLA-4 expression and IL-10 production of CD4^＋CD25^＋＋ Tregs in vivo.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2010年第1期29-31,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30872683、30971192）
国家重点基础研究发展规划项目（2005CB522602）
