血红素加氧酶-1上调CD4^+ CD25^+ Treg foxp3表达以增强Treg的免疫抑制功能  

Heme oxygenase-1 upregulates the expression of CD4^+ CD25^+ regulatory T cells foxp3 to enhance of the immunosuppressive function of these cells

作　　者：须丽清[1] 钟文伟[1] 李宁丽[2] 邵洁[1] 李云珠[1] 俞善昌[1] 夏振炜[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院儿科,上海200025 [2]上海交通大学医学院上海市免疫学研究所,上海200025

出　　处：《现代免疫学》2006年第6期469-476,共8页Current Immunology

基　　金：国家自然科学基金资助项目(30170988;30570798);上海市科委资助项目(044119662);上海市教委资助项目(03BZ04)

摘　　要：本研究探讨血红素加氧酶-1(heme oxygenase-1,HO-1)诱导CD4+CD25+调节性T细胞(regulatory T cells,Treg)foxp3表达,增加IL-10分泌,提高CD4+CD25+Treg的免疫抑制功能。选用磁珠分离正常BALB/c小鼠脾脏CD4+CD25+Treg,转染含HO-1质粒pcDNA3HO-1,或用血红素(hemin)、锡-原卟啉(Sn-protoporphyrin,SnPP)干预,培养48 h。用卵清蛋白致敏、激发BALB/c小鼠建立哮喘模型,并在致敏、激发阶段分别经血红素和SnPP干预。用Real-Time PCR和Western blot方法分别测定培养细胞内HO-1、foxp3 mRNA及蛋白量;ELISA方法分别测定细胞上清液和动物血清中IL-10、TGF-β水平;用磁珠分离哮喘动物脾脏CD4+CD25+Treg进行功能抑制试验。结果显示:经pcDNA3HO-1和血红素上调CD4+CD25+Treg HO-1表达,foxp3表达及蛋白水平相应增加,上清液IL-10水平明显升高。而OVA致敏、激发的哮喘小鼠模型,经血红素干预后,血清IL-10分泌亦增多,CD4+CD25+Treg功能抑制作用显著增强。该结果表明HO-1诱导CD4+CD25+Treg特异性转录因子foxp3表达,促进IL-10分泌,增强CD4+CD25+Treg的调节功能,具有显著的免疫抑制作用。TO explore the expression of CD4^＋ CD25^＋ regulatory T cells （Treg） foxp3 and the increase of IL-10 secretion induced by heme oxygenase-1 （HO-1） to enhance Treg immunosuppressive function. CD4^＋CD25^＋ Treg from BALB/c mouse spleen were negatively isolated by microbeads, and the cells were transfected by plasmids pcDNA3 and pcDNA3HO-1 or treated with hemin and Sn-protoporphyrin （SnPP）, respectively. BALB/c mouse was sensitized and challenged by ovalbumin （OVA） and administrated with hemin or SnPP. The expression levels of HO-1, foxp3 mRNA and the proteins production were analyzed in CD4^＋ CD25^＋ Treg, and concentration of IL-10 as well as TGF-β in culture supernatants and serum were measured. Then the suppressive capacity of CD4^＋ CD25^＋ Treg was detected. The results showed that the expression levels of foxp3 mRNA and the protein production were enhanced after CD4^＋ CD25^＋ Treg transfected by pcDNA3HO-1 and treatment with hemin, the concentration of IL-10 was increased in culture supernatant and in serum of BALB/c mouse sensitized and challenged by OVA and herain. The suppressive capacity of CD4^＋ CD25^＋ Treg was also increased in group of mice treated with hemin. This study indicates that HO-1 induced the upregulation of CD4^＋CD25^＋ Treg foxp3 mRNA and the protein production, promote the excretion of IL-10, and enhance immunosuppressive function.

关 键 词：血红素加氧酶 foxp3^+CD4^+CD25^+调节性T细胞 IL-10 哮喘 

分 类 号：R392.12[医药卫生—免疫学]