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目的:探究γδT细胞在急性铜绿假单胞菌肺炎宿主早期天然免疫中的作用。方法:建立急性铜绿假单胞菌肺部感染小鼠模型,分别设野生对照组、野生感染组、免疫球蛋白(Ig)G感染组、抗γδT细胞受体(TCR)感染组,应用流式细胞术测定产白介素(IL)-17-γδT细胞在肺内的表达。应用抗γδTCR抗体耗竭小鼠体内γδT细胞,通过实时定量PCR和酶联免疫吸附试验(ELISA)法测定肺内IL-17A、IL-17F mRNA和蛋白水平的表达,并评估肺内细菌负荷以及病理改变。结果:1在急性铜绿假单胞菌肺部感染8 h后,小鼠肺内产IL-17-γδT细胞的表达明显增加。2小鼠在感染8 h后,抗γδTCR组小鼠肺内IL-17A的mRNA和蛋白水平较IgG对照组明显下降(P<0.01);抗γδTCR组小鼠肺内IL-17F的mRNA和蛋白水平与IgG对照组相比均无明显升高;抗γδTCR组肺泡灌洗液(BALF)中细胞分类提示中性粒细胞明显减少。3小鼠在感染16 h后,抗γδTCR组肺内细菌负荷仍明显存在,是IgG对照组的174倍,其病理提示炎症反应依然明显存在。结论:在急性铜绿假单胞菌肺部感染宿主天然免疫早期,γδT细胞是IL-17A的主要来源细胞,且起着协助病原菌清除的积极作用。
Objective: To investigate the role of γδT cells in the early innate immunity of acute Pseudomonas aeruginosa pneumoniae host. Methods: Acute pulmonary infection in mice with Pseudomonas aeruginosa was established. Wild control group, wild-type infection group, immunoglobulin (G) infection group and anti-γδ T cell receptor (TCR) infection group were established. Cytometry was used to determine the expression of interleukin (IL) -17-γδT cells in the lung. Γδ T cells were depleted in mice by anti-γδTCR antibody. The expression of IL-17A, IL-17F mRNA and protein in the lungs was measured by real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA) change. Results: 1 After acute pulmonary infection of Pseudomonas aeruginosa for 8 h, the production of IL-17-γδT cells in the lung of mice significantly increased. 2 mRNA and protein levels in the lungs of mice with anti-γδTCR were significantly lower than those in the IgG control group (P <0.01) after 8 h of infection. The mRNA and protein expressions of IL-17F There was no significant difference in the level of neutrophils between the control group and the cell sorting in the anti-γδTCR BALF. 3 mice in the anti-γδTCR group after 16 h infection was still evident in the lung bacterial load, which is 174 times the IgG control group, the pathology suggests that the inflammatory response is still evident. Conclusion: In the early stage of innate immunity of acute P. aeruginosa lung infection in host, γδT cells are the main source cells of IL-17A, and play an active role in assisting the clearance of pathogens.