哮喘大鼠树突状细胞CCR7对共刺激分子CD80、CD86表达的影响研究  

Effect of dendritic cells CCR7 on the expression of co-stimulatory molecules CD80 and CD86 in asthmatic rats

作　　者：逯丹 李毅[2] 杜永成[2] 刘丽萍[1] 郎依 LU Dan;LI Yi;DU Yongcheng;LIU Liping;LANG Yi(The Fifth Clinical Medical College of Shanxi Medical University,Taiyuan,Shanxi 030012,China;Department of Respiratory and Critical Medicine,The Fifth Hospital of Shanxi Medical University,Taiyuan,Shanxi 030012,China)

机构地区：[1]山西医科大学第五临床医学院,山西太原030012 [2]山西医科大学第五医院呼吸与危重症医学科,山西太原030012

出　　处：《临床肺科杂志》2024年第9期1368-1373,共6页Journal of Clinical Pulmonary Medicine

基　　金：山西省回国留学人员重点科研资助项目(No.2014-重点8)。

摘　　要：目的 通过观察哮喘大鼠树突状细胞CCR7对共刺激分子CD80、CD86表达的影响,研究其在免疫耐受中的作用。方法 建立哮喘大鼠模型,提取骨髓来源的树突状细胞,将编码过表达CCR7基因的重组腺病毒及有靶向CCR7基因的短发夹RNA腺病毒载体转染树突状细胞(DC)。同时培养脾脏来源T淋巴细胞,与转染后的DC行混合淋巴细胞反应,采用MTT比色法测定淋巴细胞增殖能力;采用蛋白质免疫印迹试验检测DC中CCR7的蛋白表达水平;流式细胞仪检测DC表面CD80、CD86表达;酶联免疫吸附法检测IL-10、IL-12、TGF-β表达水平。结果 CCR7基因过表达组CCR7蛋白表达水平增高,CD80、CD86表达水平较哮喘组明显升高,CCR7基因敲低组二者的表达水平较哮喘组降低(P<0.001);IL-12在CCR7基因敲低组较哮喘组明显升高;CCR7基因过表达组IL-10及TGF-β表达水平较敲低组升高,CCR7基因敲低组TGF-β蛋白表达水平较哮喘组明显降低(P<0.001)。结论 共刺激分子的激活可能通过DC表面CCR7表达的介导来实现。同时,CCR7可影响IL-10、IL-12和TGF-β的蛋白表达水平来诱导哮喘免疫耐受。Objective To reveal the effects of CCR7 from dendritic cells(DC)on the expression of co-stimulatory molecules CD80 and CD86 in asthmatic rats.Methods An Asthma rat model was established,and DC derived from bone marrow was extracted.In the present study,bone marrow-derived DC was transfected with an adenovirus encoding the rat gene CCR7 or a short hairpin RNA targeting CCR7(shCCR7).Meanwhile,spleen-derived T cells were cultured and mixed with transfected DC,and the proliferation ability of immune cells was measured by the MTT method.The protein expression level of CCR7 on the DC surface was detected by Western blot assay.The expressions of CD80 and CD86 on the DC surface were detected by flow cytometry(FACS).The expression of IL-10,IL-12,and transforming growth factorβ(TGF-β)were detected by enzyme-linked immunosorbent assay(ELISA).Results The expression of CCR7 in the CCR7 over-expression group was significantly increased,the expression of CD80 and CD86 were significantly increased compared with the asthma group,and the expression of gene CCR7 knockdown group was decreased compared with the asthma group(P<0.001).IL-12 level in the gene CCR7 knockout group was significantly higher than that in the asthma group.The level of IL-10 and TGF-βin the CCR7 over-expression group was increased compared with the gene CCR7 knockdown group,and the level of TGF-βin the knockdown group was decreased compared with the asthma group(P<0.001).Conclusion The activation of costimulatory molecules may be mediated by the expression of CCR7 on the DC surface.At the same time,CCR7 can affect the expression of IL-10,IL-12,and TGF-βto induce asthma immune tolerance.

关 键 词：支气管哮喘 CCR7 共刺激分子 免疫耐受 

分 类 号：R562.25[医药卫生—呼吸系统]