细胞芯片检测新生隐球菌菌株丝氨酸蛋白酶的表达

Cell microarray in detection of serine protease expression in Cryptococcus neoformans

目的:利用细胞芯片技术检测新生隐球菌菌株中丝氨酸蛋白酶的表达,初步探讨丝氨酸蛋白酶在新生隐球菌致病过程中的作用。方法:不同来源和血清型的新生隐球菌菌株36株,应用组织芯片构建仪制备菌株芯片,利用菌株细胞芯片和免疫组化技术对菌株中丝氨酸蛋白酶的表达情况进行检测。结果:丝氨酸蛋白酶在所有菌株细胞中强阳性表达率为67.0%(25/36),强阳性表达率在血清A型、血清B型、血清D/AD型分别为46.2%(6/13)、92.3%(12/13)、66.7%(4/6),在环境分离株、临床分离株及荚膜缺陷株中分别为55.6%(5/9)、82.6%(19/23)、25%(1/4)。不同血清型及不同来源菌株中,以血清B型、临床分离株中的丝氨酸蛋白酶阳性表达率高(P<0.05)。结论:菌株细胞芯片是检测致病性真菌样本成分的新技术;致病力强的新生隐球菌临床分离株中的丝氨酸蛋白酶表达最高,提示丝氨酸蛋白酶在临床菌株致病过程中起到主要作用。

Objective：To determine the serine protease expression in Cryptococcus neoformans （C. neoformans） by cell microarray technique,so as to investigate the role of serine protease in the pathogenesis of C. neoformans. Methods： The cell microarray was constructed with tissue microarray. Thirty-six strains of C. neoformans of different sources and homologous serotypes were examined for their serine protease expression by cell microarray technique and immunohistochemistry. Results： Strong expression of serine protease was found in 25 （67.0 % ） strains. The rates of strong serine protease expression in serotype A, B and D/AD strains were 46.2% （6/13）,92.3% （12/13）and 66.7%（4/6）, and in environment-isolated,clinically isolated and uncapsuled strains were 55.6 % （5/9）, 82.6 % （19/23） and 25 % （ 1/4）, respectively. Serine protease expressions in serotype B and clinically-isolated strains were significantly higher than that in other group （P〈0.05）. Conclusion： Microarray of strain cells is a new method for identifying pathogenic fungus. Higher expression of serine protease in clinically-isolated strains is associated with strong virulence of clinical isolates strains, suggesting that serine protease plays a major role in the pathogenesis of C. neo f ormans.