炭疽芽孢在巨噬细胞RAW264.7内的萌发过程

Germination process of Bacillus anthracis endospores within macrophage RAW264.7

目的:确定炭疽杆菌A16R芽孢在巨噬细胞系RAW264.7 内萌发的过程. 方法:用炭疽杆菌A16R芽孢以MOI 20:1感染RAW264.7,分别于吞噬后1、2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、7、7.5、8 h取细胞爬片进行复红美蓝染色,显微镜观察. 结果:吞噬后2-2.5 h,细胞内的芽孢开始萌发并形成繁殖体;吞噬后3.5-4 h,繁殖体开始二分裂;吞噬后5-5.5 h, 繁殖体进入指数增长阶段;吞噬后7-8 h,大量的繁殖体充斥于细胞内、外,细胞开始崩解. 结论:首次采用复红美蓝染色确定了A16R炭疽芽孢在巨噬细胞RAW264.7内的萌发过程.

AIM: To confirm the germination process of Bacillus anthracis A16R endospores within murine macrophage RAW264.7 METHODS: Macrophage RAW264.7 cells were infected by Bacillus anthrax A16R (pXO2-) spores at a multiplicity of infection (MOI) of 20:1. Then the cells were harvested at different time points (1, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 and 8 h after infection). The growth of infected cells was observed under light microscope by fuchsin basic methylene blue staining. RESULTS: The endospores began to germinate and develop into vegetative bodies 2 to 2.5 hours after infection. The vegetative bodies entered the phase of binary fission 3.5 to 4 hours after infection. At 5 to 5.5 hours, the bacillus proliferated into exponential phase andthe macroph-ages began to lyse 7 to 8 hours after infected. CONCLUSION: For the fisrt time, fuchsin basic methylene blue staining is used to study the germination process of Ba-cillus anthracis endospores within macrophage RAW264.7.