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目的通过比较钩端螺旋体(钩体)在人和小鼠单核-巨噬细胞内的存活情况,探讨固有免疫在钩端螺旋体病致病机制中的作用。方法将钩体秋季血清群强毒株56606v株及其经体外多次传代的弱毒株56606 a株在体外分别感染经佛波酯(PMA)诱导分化的人单核细胞系THP-1和小鼠单核-巨噬细胞系RAW 264.7。感染后2、24、72 h,采用免疫荧光染色激光共聚焦显微镜观察并计算两种细胞的含菌细胞百分数;感染后2、12、24、48和72 h,采用Real-Tim e PCR技术定量检测可间接反映细胞内钩体存活数量的钩体16S rRNA的表达。结果随着感染时间的延长,两种细胞的含菌细胞百分数逐渐降低;Real-Tim e PCR检测结果显示:随着感染时间的延长,两种细胞内钩体16S rRNA的表达逐渐下调,细胞内钩体的存活数量逐渐减少。结论钩体在人和小鼠单核-巨噬细胞内均不能存活和繁殖,提示致病性钩体通过抗巨噬细胞的杀灭降解而在胞内的存活和繁殖可能不是钩体感染人类宿主的主要致病途径,其发病机制有待进一步阐明。
Objective To investigate the role of innate immunity in the pathogenesis of leptospirosis by comparing the survival of leptospira in human and mouse monocytes with macrophages. Methods The attenuated strain 56606v and the attenuated strain 56606a, which were infected by PMA, were inoculated with the attenuated strain 56606v and the attenuated strain 56606a, respectively, in vitro to infect the human monocytic cell line THP-1 and mouse Monocyte - Macrophage cell line RAW 264.7. At 2, 24, 72 h after infection, the percentage of cells containing bacteria in the two cells was observed by immunofluorescence staining and confocal laser scanning microscopy. Real-time PCR was used to quantify the cells at 2, 12, 24, 48 and 72 h Detection of the expression of 16S rRNA of Leptospira, which indirectly reflects the number of intraocular leptospirosis. Results With the extension of infection time, the percentages of bacteria-containing cells in both cells decreased gradually. The results of Real-Tim e PCR showed that the expression of 16S rRNA in both cells decreased gradually with the extension of infection time, The number of hook body gradually decreased. Conclusions Lysosomes can not survive or multiply in both human and mouse mononuclear macrophages, suggesting that the pathogenic livers survive and multiply intracellularly by anti-macrophage killing and may not be infected by leptospirosis The main pathogenic host pathways, the pathogenesis needs further elucidation.