流式细胞仪检测胰蛋白酶对烟曲霉分生孢子表面纤连蛋白受体黏附力的影响  

The effect of trypsin on surface adhesion of fibronectin receptors of Aspergillus fumigatus conidia tested by flow cytometry

作　　者：徐赤宇[1] 王溪涛[1] 陈江汉[1] 吴建华[1] 温海[1] 

机构地区：[1]第二军医大学长征医院皮肤科,上海200003

出　　处：《中国感染与化疗杂志》2006年第1期27-29,共3页Chinese Journal of Infection and Chemotherapy

基　　金：国家自然科学基金资助项目(项目编号:30270080)

摘　　要：目的测定胰蛋白酶对真菌细胞活力的影响,探讨胰蛋白酶对烟曲霉分生孢子表面纤连蛋白受体的影响。方法用检测胰蛋白酶预处理20min后烟曲霉分生孢子的活力,检测纤连蛋白与烟曲霉分生孢子的黏附率及胰蛋白酶处理后分生孢子与纤连蛋白的黏附率的变化,同时应用激光共聚焦显微镜观察分生孢子表面纤连蛋白受体的分布情况。结果胰蛋白酶预处理的分生孢子无凋亡峰出现,被干预的分生孢子接种于马铃薯培养基后仍有大量烟曲霉生长;分生孢子与纤连蛋白的黏附作用有特异性和饱和性;定量的纤连蛋白与被干预的分生孢子的黏附率随胰蛋白酶浓度的增加而降低;干预后分生孢子表面荧光标记的纤连蛋白受体分布下降。结论蛋白水解酶类可降解真菌细胞表面相关蛋白受体;流式细胞术可成为检测真菌细胞表面微观变化的高效辅助手段之一。Objective To determine the activity of fungus cells pretreated with trypsin, and investigate the effect of trypsin on the fibronectin receptor of Aspergillus fumigatus conidia. Methods Flow cytometry was used to detect the activity of conidia after pretreatment with trypsin for 20 minutes. The binding of fibronectin to conidia and changes of adherence rate of yeast cell after pretreatment with trypsin were determined, The distribution of fibronectin-receptor on the surface of conidia was observed by immunofluorescence confocal microscopy. Results Apoptosis peak was not observed in the yeast cells pretreated with trypsin. Dense A. fumigatus colonies were present after the pretreated conidia were inoculated on PDA. Binding of fibronectin to conidia was specific and saturable, The adherence ratio of quantitative fibronectin to the pretreated conidia decreased as tryp sin concentration increased. The Fn-receptors labeled with FITC reduced on the surfaces of yeast cell after pretreatment with trypsin. Conclusions Proteolytic enzymes can degrade the protein receptors on the surface of yeast cells. FCM can be a highly reliable way to detect the microcosmic change of fungal cell surface.

关 键 词：烟曲霉 流式细胞术 胰蛋白酶 蛋白水解酶 

分 类 号：R379[医药卫生—病原生物学]