钙荧光白和荧光刀豆蛋白A标记的白念珠菌生长活性和粘附能力

Growth and Adhesion to Cells of Candida albicans Labeled by Calcofluor White and Fluorescent Concanavalin A

【目的】探讨钙荧光白和荧光刀豆蛋白A标记白念珠菌后对其生长活性和粘附能力的影响,以明确在念珠菌和宿主细胞相互作用研究中钙荧光白和荧光刀豆蛋白A是否适用于活念珠菌的标记。【方法】分别使用钙荧光白和结合Alexa fluor 488的刀豆蛋白A标记20株白念珠菌,将标记荧光的白念珠菌酵母细胞接种至酵母提取物蛋白胨葡萄糖培养基,通过菌落计数法检测荧光标记的白念珠菌的生长活性。将荧光标记的白念珠菌酵母细胞分别与巨噬细胞和肠上皮细胞共培养,使用荧光显微镜检测白念珠菌的粘附能力。【结果】钙荧光白组和荧光刀豆蛋白A组白念珠菌的菌落数目和未标记的对照组相比,均无统计学差异(P=0.942,P=0.885);钙荧光白组和荧光刀豆蛋白A组的白念珠菌在巨噬细胞上的粘附数目均比对照组少(P=0.000,P=0.000);钙荧光白组和荧光刀豆蛋白A组的白念珠菌在肠上皮细胞上的粘附数目也均比对照组组少(P=0.000,P=0.000)。对照组组白念珠菌与细胞共培养半小时后可长出菌丝,而钙荧光白标记的白念珠菌与细胞共培养半小时后未长出菌丝。【结论】钙荧光白和荧光刀豆蛋白A标记白念珠菌后不影响其生长活性,可显著降低其粘附巨噬细胞和肠上皮细胞的能力。钙荧光白染色影响白念珠菌的出芽过程。钙荧光白和荧光刀豆蛋白A不适用活念珠菌的标记。

【Objective】To assess effect of calcofluor white and fluorescent dye labeled concanavalin A in Candida albicans growth and adhesion of cells.【Methods】Yeast cells of 20 strains of Candida albicans were labeled by calcofluor white and Alexa fluor 488 conjugated concanavalin A respectively. The growth of labeled Candida albicans were tested by counts of colony forming unit. Then yeast cells of Candida albicans were co-cultured with macrophages and enterocytes for half an hour. The adhesion of labeled Candida albicans to macrophages and enterocytes were observed by fluorescence microscopy.【Results】No difference was observed on the number of colony forming units between calcofluor white-labeled groupand control group(P=0.942). Also,no difference was observed on the number of colony forming units between Alexa fluor 488 conjugated concanavalin A-labeled group and control group. However,either the number of calcofluor white-labeled Candida albicans or Alexa fluor 488 conjugated concanavalin A-labeled Candida albicans that bound to macrophages was less than that in control group(P=0.000,respectively). Either the number of calcofluor white-labeled Candida albicans or Alexa fluor 488 conjugated concanavalin A-labeled Candida albicans that bound to enterocytes was less than that in control group(P = 0.000,respectively). Hyphae were observed in control group but not in calcofluor white group after yeast cells of Candida albicans were co-cultured with cells for half an hour.【Conclusions】Growth of Candida albicans was not changed,while its adhesion to cells was reduced after its labeling by calcofluor white and Alexa fluor 488 conjugated concanavalin A.The germination of Candida albicans was affected when it had been labeled by calcofluor white.