过继经母牛分枝杆菌雾化免疫后的γδT细胞对哮喘小鼠模型的影响及其机制  

Effect and mechanism of γδT cells immunized by adoptive Mycobacterium vaccae atomiza⁃tion on a mouse model of asthma

作　　者：黄仲辰 陈晓菊[2] 姚意恩 李超乾[1] Huang Zhongchen;Chen Xiaoju;Yao Yien;Li Chaoqian(Department of Respiratory Medicine,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Department of Comprehensive Internal Medicine,The Affiliated Cancer Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院呼吸内科,南宁530021 [2]广西医科大学附属肿瘤医院综合内科,南宁530021

出　　处：《广西医科大学学报》2022年第1期7-12,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81470230)。

摘　　要：目的:探讨过继经母牛分枝杆菌雾化免疫后的γδT细胞对哮喘小鼠模型的影响及其机制。方法:制备TCR^(-β-/-)小鼠分别经生理盐水和母牛分枝杆菌雾化后γδT细胞悬浮液;将32只无特定病原体(SPF)级雄性Balb/c小鼠随机均分为A组、B组、C组、D组,A组为阴性对照组,B组为哮喘模型对照组,C组为过继经生理盐水雾化后的γδT细胞对照组,D组为过继经母牛分枝杆菌雾化免疫后的γδT细胞预防组;B组、C组、D组均用鸡卵清蛋白(OVA)常规制备哮喘模型,C组、D组都在第1次腹腔注射OVA前10 min及第1次雾化OVA激发前10 min经鼠尾静脉分别注射1×10^(6)个γδT细胞;A组腹腔注射及雾化均用PBS代替;末次雾化后24 h内检测气道反应性,处死小鼠,行支气管肺泡灌洗收集肺泡灌洗液(BALF),酶联免疫吸附测定实验(ELISA)检测BALF的IL-13和IL-5的水平,取肺组织做苏木精—伊红(HE)和过碘酸—雪夫(PAS)染色,免疫组织化学实验检测肺组织中T-bet/GATA3蛋白的表达。结果:B组小鼠气道反应性高于A组、D组(P<0.05),D组小鼠气道反应性低于B组(P<0.05),C组小鼠气道反应性相比B组变化不明显;相对比于B组、C组,D组支气管周围炎症细胞浸润减少,D组管腔中杯状细胞较B组、C组减少;B、C组中肺组织的IL-13、IL-5表达高于A、D组(P<0.05);B组、C组肺组织的GATA3表达明显高于A组、D组(P<0.05),T-bet表达低于A、D组(P<0.05)。结论:过继经母牛分枝杆菌雾化免疫后的γδT细胞对哮喘小鼠模型具有预防作用,可能与分别上调或下调T-bet、GATA-3水平,以及调节IL-13、IL-5分泌有关。Objective:To investigate the effect ofγδT cells after adoptive immunization with mycobacterium vaccae atomization on asthma mouse model and its mechanism.Methods:TCR-β-/-mice were atomized with normal saline and mycobacterium vaccae respectively,and theγδT cell suspension was obtained.Thirty-two SPF BALB/c male mice were randomly divided into groups A,B,C,and D:group A was negative control group,group B was asthma model control group,group C was adoptedγδT cells atomized with normal saline control group,and group D was adoptedγδT cells prevention group after atomized immunization with Mycobacteria vaccae.Asthma models in group B,C and D were prepared with chicken ovalbumin(OVA).A total of 1×10^(6)γδT cells were injected through tail vein 10 min before the first intraperitoneal injection of OVA and 10 min before the first atomization of OVA,respectively,in group C and D.PBS was used to replace intraperitoneal injection and atomization in group A.Airway reactivity was detected within 24 hours after the last atomization.The mice were sacrificed and bronchoalveolar lavage was performed to collect alveolar lavage fluid(BALF).IL-13 and IL-5 levels in BALF were detected by ELISA.Lung tissues were stained with HE and PAS.The expression of T-bet/GATA3 protein in lung tissues was detected by immunohistochemistry.Results:The airway reactivity in group B was significantly higher than that in group A and D(P<0.05).The airway reactivity was similar in group BandC.Compared with group B and C,the infiltration of inflammatory cells around bronchus in group D was significantly reduced.The number of goblet cells in the lumen of group D was significantly lower than that of group B and C.The expression of IL-13 and IL-5 in lung tissues of group B and C was significantly higher than that of group A and D(P<0.05).The expression of GATA3 in lung tissues of group B and C was significantly higher than that of group A and D,while the expression of T-bet was significantly lower(P<0.05).Conclusion:Adoptive immunization with Mycob

关 键 词：哮喘 母牛分枝杆菌 T-BET GATA-3 ΓΔT细胞 IL-13 IL-5 

分 类 号：R562.25[医药卫生—呼吸系统]