BTLA在重症社区获得性肺炎患者中的表达及糖皮质激素干预动物模型的影响  

BTLA expression in patients with severe community-acquired pneumonia and the impact of glucocorticoid intervention in animal models

作　　者：周国旗[1] 王导新[2] 刘代顺[1] 刘振峰[1] 李朝英[1] 

机构地区：[1]遵义市第一人民医院呼吸内科,563000 [2]重庆医科大学第二附属医院呼吸内科,400010

出　　处：《免疫学杂志》2017年第4期312-320,共9页Immunological Journal

基　　金：贵州省卫计委科学技术基金(2016-1-018);遵义市科技计划(遵市科合社〔2016〕04号)

摘　　要：目的本研究通过观察BT淋巴衰减子(B and T lymphocyte attenuator,BTLA)在重症CAP患者外周血淋巴细胞及支气管黏膜上的表达,外源性干预LPS-诱导的急性肺部炎症小鼠模型BTLA表达对肺部炎症的作用以及给予糖皮质激素对BTLA表达的影响。方法利用流式细胞学分析重症CAP患者外周血BTLA^+CD4^+淋巴细胞比例,另外对重症CAP患者支气管黏膜常规行HE染色和免疫组织化学检测BTLA表达情况。利用流式细胞学分析LPS诱导的急性肺部炎症模型右心血BTLA^+CD4^+淋巴细胞比例。收集肺泡灌洗液(bronchoalveolar lavage fluid,BALF)行炎症细胞分类计数,并利用ELISA检测炎症因子。取肺组织分别行HE染色、免疫组织化学染色和蛋白印迹。结果流式细胞学分析重症肺炎患者外周血中BTLA^+CD4^+淋巴细胞比例明显高于正常对照组。此外,支气管黏膜HE染色结果显示重症CAP患者支气管黏膜有大量炎症细胞浸润,并且伴有支气管壁增厚;进一步用免疫组织化学检测支气管黏膜BTLA表达,结果提示重症肺炎患者支气管黏膜BTLA表达明显增高,浸润的炎症细胞BTLA表达增高明显。流式细胞学分析LPS诱导的急性肺部炎症模型小鼠右心血中BTLA^+CD4^+淋巴细胞比例明显高于正常对照组。利用激动性抗-BTLA抗体6A6干预急性肺部炎症模型后淋巴细胞BTLA表达较模型小鼠明显增高,同时,激动性抗-BTLA抗体6A6能减轻肺部炎症反应和抑制NF-κB信号通路激活。利用糖皮质激素干预急性肺部炎症模型后淋巴细胞BTLA表达较模型小鼠明显增高,糖皮质激素能减轻肺部炎症反应和抑制NF-κB信号通路激活。结论通过本研究我们推测BTLA信号可能参与了重症CAP患者的免疫调节过程,BTLA可能成为重症CAP治疗的一个新靶点。同时,BTLA的表达可能反应患者机体的免疫状态,为临床使用糖皮质激素治疗提供指导作用。In this study, we investigated potential changes in B and T lymphocyte attenuator（BTLA） expressedon lymphocytes and bronchial mucosa in patients with severe community acquired pneumonia（CAP）, and toevaluate the effects of BTLA on a model of LPS-induced acute lung inflammation. The ratio of BTLA^＋CD4^＋lymphocytes in human subject was analyzed by flow cytometry; HE staining and immunohistochemical staining wereemployed to detecl the BTLA in bronchial mucosa from patients with severe CAP. The percentages ofcirculating BTLA^＋CD4^＋lymphocytes were determined in an LPS-induced acute lung inflammation model.Bronchoalveolar lavage fluid（BALF） was collected from mice and analyzed for leukocyte counts by enzyme-linkedwere collected and assessed via Western blotting,immunohistochemistry and HE staining. Dataowedthat the percentage of circulating BTLA^＋ shCD4^＋lymphocytes in patients with severe CAP wassignificantly higher than that in control groups; HE staining showed that there were inflammatory cell infiltration inbronchial mucosa and submucosa, as well as thickening of bronchial walls in patients with severe CAP.Immunohistochemistry for BTLA in bronchial mucosa showed that BTLA protein expression was upregulated inbronchial epithelium and inflammatory cells of patients with severe CAP. The percentage of circulating BTLA^＋CD4^＋lymphocytes were significantly higher in the LPS-induced acute lung inflammation model than that in control groups.Agonistic anti-BTLA antibody 6A6 could upregulate BTLA expression significantly in LPS-induced acute lunginflammation mice. Moreover, the agonistic anti-BTLA antibody 6A6 attenuated inflammatory responses and NF-κBpathway activation in model mice. These results demonstrated that BTLA play an important role in immunoregulationof severe CAP, which may be a therapeutic target for the treatment of severe CAP. Meanwhile, BTLA expression mayreflect the body＇s immune status and guide decisions regarding steroid therapy for treating severe CAP.

关 键 词：社区获得性肺炎 BT淋巴衰减子 糖皮质激素 脂多糖 

分 类 号：R563.1[医药卫生—呼吸系统]