绵羊布鲁氏菌侵染小鼠巨噬细胞的荧光表征与分析

通过对绿色荧光蛋白(GFP)布鲁氏菌16M(强毒株)和M5(弱毒株)侵染小鼠巨噬细胞及胞内溶酶体、内质网、高尔基体初次结合所用时间进行测定,探讨二者在结合所用时间上是否存在差异;其次比较GFP布鲁氏菌16M和M5与胞内各细胞器结合产生的荧光强度。将GFP布鲁氏菌16M(强毒株)和M5(弱毒株)分不同时间段分别侵染RAW 264.7(细菌和细胞比例为100︰1),利用激光共聚焦显微镜和流式细胞仪观察和检测。结果表明:布鲁氏菌16M和M5侵染初期10 min时进入巨噬细胞,1.5 h时布鲁氏菌到达溶酶体,2.0 h时布鲁氏菌到达内质网和高尔基体。结果提示,布鲁氏菌16M和M5在侵染进入宿主细胞与胞内各细胞器初次结合所用时间相同,但布鲁氏菌16M与各细胞器结合的荧光强度均高于布鲁氏菌M5。以此推断布鲁氏菌16M进入胞内的数量高于布鲁氏菌M5。

The Fluorescent Characterization and Analysis of Macrophages Infected by B.melitensis

The fist combining time of GFP Brueella 16M （virulent strain） and M5 （weak strain） infecting mice macrophage, lysosome,endoplasmic reticulum and Golgi body of the macrophage was measured so that to explore if there are time-length differences of the combing time,and the fluorescence intensities produced by intracellular organelles combining with GFP Brueella 16M and M5 were eompared.GFP Brueella 16M and M5 infected mice maerophage RAW 264.7 in different time lengths of the their combiation,the ratio of bacteria and cell ratio was 100：l,and the results were observed and detected by confocal laser scanning microscope and flow cytometry.The results showed that Brucella 16M and M5 gained entry to macrophages in 10 min,lysosomes in 1.5 h,endoplasmic reticulum and Golgi body in 2.0 h.But the fluorescence intensities produced by intracellnlar organelles combining with GFP Brucella 16M was higher than M5.The results suggested that Brucella 16M and M5 eombining with the organelles spent the same time.So we can infer the number of Brucella 16M entering into macrophages was higher than M5.