论文部分内容阅读
目的探讨小鼠腹腔巨噬细胞对马尔尼菲青霉酵母相细胞的影响,为研究抗马尔尼菲青霉的免疫机制提供一定的实验依据。方法将马尔尼菲青霉酵母相细胞与小鼠腹腔巨噬细胞在体内和体外两种方式下进行共培养,分别于60min、120min、240min和360min后进行菌落形成单位(CFU)计数,统计分析体内组与体外组CFU数结果的差异性。利用钙荧光白染色巨噬细胞胞内的马尔尼菲青霉酵母相细胞,荧光显微镜下观察细胞形态特征的变化。结果经体内和体外两种方式共培养,两组之间的CFU计数差异无统计学意义,但两组内不同时间点的CFU计数差异存在统计学意义。荧光显微镜下可见巨噬细胞内的酵母细胞被钙荧光白染上淡蓝色荧光。结论小鼠腹腔巨噬细胞在体内、外对马尔尼菲青霉酵母相细胞不起明显的杀灭或溶解作用,其最大的吞噬时间为共培养240min。钙荧光白染色可快速有效的区分巨噬细胞与真菌,是一种良好的检测真菌方法。
Objective To investigate the effect of mouse peritoneal macrophages on the phase cells of Penicillium marneffei yeast and to provide some experimental evidence for studying the immune mechanism against Penicillium marneffei. Methods The Penicillium marneffei phase cells and mouse peritoneal macrophages were cocultured in vitro and in vivo. The number of colony forming unit (CFU) was counted after 60min, 120min, 240min and 360min, respectively. Statistical analysis Differences of CFU results between in vivo and in vitro groups. Macrophages of Penicillium marneffei were stained with calcium fluorescein to observe the morphological changes of cells. The results were co-cultured in vitro and in vivo. There was no significant difference in CFU count between the two groups. However, there was significant difference in CFU count between two groups at different time points. Fluorescence microscopy showed that macrophages in yeast cells were fluorescently labeled with calcium-fluorescein blue light. Conclusion Mice peritoneal macrophages can not obviously kill or dissolve Penicillium marneffei phage cells in vivo and in vitro. The maximal phagocytosis time is 240 min. Calcium Fluorescent staining can quickly and effectively distinguish macrophages and fungi, is a good method to detect fungi.