TIM-1-Fc融合蛋白对哮喘小鼠Th1/Th2和Th17/Treg免疫失衡的调节  

TIM-1-Fc Fusion Protein Regulates Th1/Th2 and Th17/Treg Immune Imbalance in Mice with Asthma

作　　者：曹津萌 卿吉琳[2] 朱莉雅 魏燕 赵艺莲 叶超 陈治中[5] Cao Jinmeng;Qing Jilin;Zhu Liya(School of Clinical Medicine,Guilin Medical University,Guilin 541199,China;Reproductive Medicine Center,People's Hospital of Guangxi Zhuang Autonomous Region,Nanning 530021,China;Gradua te School,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]桂林医学院临床医学院,桂林541199 [2]广西壮族自治区人民医院生殖医学中心,南宁530021 [3]广西中医药大学研究生院,南宁530200 [4]广西医科大学第一临床医学院,南宁530021 [5]广西壮族自治区人民医院精准联合检验中心,南宁530021

出　　处：《华中科技大学学报（医学版）》2024年第4期479-486,527,共9页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81960428,No.U22A2092,No.81260007)。

摘　　要：目的制备T细胞免疫球蛋白和黏蛋白结构域1(T cell immunoglobulin domain and mucin domain protein-1,TIM-1)-Fc融合蛋白并探讨TIM-1-Fc融合蛋白对卵白蛋白(ovabumin,OVA)诱导的哮喘小鼠的干预作用及潜在作用机制。方法通过基因工程技术获得TIM-1-Fc融合蛋白。采用腹腔注射OVA氢氧化铝溶液致敏建立过敏性哮喘小鼠模型,随机分为对照组、哮喘组和TIM-1-Fc干预组。每次干预前20 min,使用40μL卵白蛋白生理盐水溶液(OVA-NS)滴鼻,TIM-1-Fc融合蛋白干预包括TIM-1-Fc滴鼻组(每只小鼠每次给予1μg/40μL TIM-1-Fc滴鼻)和TIM-1-Fc注射组(每只小鼠每次给予6μg/200μL TIM-1-Fc腹腔注射),每天1次,连续7 d,对照组用生理盐水替代。采用苏木精-伊红(HE)染色观察肺组织病理变化;采用流式细胞术检测小鼠外周血中辅助性T细胞2(type 2 T helper cells,Th2)、辅助性T细胞17(type 17 T helper cells,Th17)和调节性T细胞(Treg)比例及相关细胞因子水平。结果成功构建TIM-1-Fc融合蛋白,成功构建OVA诱导的过敏性哮喘小鼠模型。与哮喘组相比,TIM-1-Fc融合蛋白干预后显著减轻了哮喘小鼠气道炎性损伤和肺组织损伤;TIM-1-Fc融合蛋白干预能显著降低外周血中TIM-1^(+)CD4^(+)T细胞和TIM-1^(+)Th17细胞比例,使TIM-1^(+)Treg细胞增多,显著降低Th2、Th17细胞比例,提高Treg细胞比例,调节哮喘中Th1/Th2和Th17/Treg免疫失衡。结论TIM-1-Fc融合蛋白改善OVA诱导的过敏性哮喘小鼠气道炎症和肺组织损伤,其作用机制可能与TIM-1-Fc融合蛋白对Th1/Th2和Th17/Treg的免疫调节有关。Objective To prepare T cell immunoglobulin domain and mucin domain 1 protein-1(TIM-1)-Fc fusion protein,and to investigate the intervention effect of TIM-1-FC fusion protein on ovabumin(OVA)-induced asthma mice as well as its potential mechanism.Methods Through genetic engineering,pcDNA3.1(+)-TIM-1-Fc plasmid was constructed and stably transfected into CHO cells.Cell culture supernatant was collected and purified,and TIM-1-Fc fusion protein was obtained.Mouse model of allergic asthma was established by intraperitoneal injection of OVA aluminum hydroxide solution.The mice were randomly divided into control group,asthma group and TIM-1-Fc intervention group.The TIM-1-Fc intervention group included TIM-1-Fc nasal drop group and TIM-1-Fc injection group.20 min before each intervention,40μL ovalbumin saline solution(OVA-NS)was used for nasal drops.Protein TIM-1-Fc nasal drops group(1μg/40μL TIM-1-Fc nasal drops was administrated in each mouse)and TIM-1-Fc injection group(6μg/200μL TIM-1-Fc was injected intraperitoneally in each mouse)were interfered with TIM-1-Fc fusion,once a day for 7 days.Normal saline was used as replacement in the control group.Hematoxylin-eosin staining(HE)was used to observe the pathological changes of lung tissue.Flow cytometry was used to detect the proportion of type 2 helper T cells(Th2),type 17 helper T cells(Th17),regulatory T cells(Treg)and the levels of related cytokines in peripheral blood of mice.Results TIM-1-Fc fusion protein and OVA-induced allergic asthma mouse model was successfully constructed.Compared with asthma group,TIM-1-Fc fusion protein could significantly reduce airway inflammatory injury and lung tissue injury in asthmatic mice.TIM-1-Fc fusion protein intervention could significantly reduce the proportion of TIM-1^(+)CD4^(+)T cells and TIM-1^(+)Th17 cells in peripheral blood,increase the number of TIM-1^(+)Treg cells,significantly reduce the proportion of Th2 and Th17 cells,and increase the proportion of Treg cells.It could modulate Th1/Th2 and Th17/Treg immune imb

关 键 词：TIM-1 TIM-1-Fc融合蛋白 过敏性哮喘 TH1/TH2 TH17/TREG 

分 类 号：R562.25[医药卫生—呼吸系统]