Schwienbacher Monika, Weig Michael, Thies Svenja, Regula Jörg T, Heesemann Jürgen, Ebel Frank
Max-von-Pettenkofer-Institute, Ludwig-Maximilans-University, Munich, Germany.
Med Mycol. 2005 Nov;43(7):623-30. doi: 10.1080/13693780500089216.
Although secreted proteins of pathogenic microorganisms often represent potential virulence factors, so far only limited information has been available on the proteins secreted by Aspergillus fumigatus. We therefore analysed supernatant proteins after growth in different media. In serum-free cell culture medium A. fumigatus growth was limited and no protein secretion was detectable, whereas distinct protein patterns were detectable after growth in either aspergillus minimal medium (AMM) or the more complex yeast glucose medium (YG). The three major proteins secreted under these conditions were identified as the ribotoxin mitogillin, a chitosanase and the aspergillopepsin i. Mitogillin and chitosanase were secreted in AMM, whereas aspergillopepsin i was especially prominent after growth in YG. When the AMM cultures reached stationary phase, seven additional major proteins were detectable. Two of them were identified as the chitinase chiB1 and a beta(1-3) endoglucanase. Conditioned medium containing mitogillin and chitosanase did not have a detectable cytotoxic effect on A549 and Vero cells. Using recombinant mitogillin and chitosanase we detected anti-chitosanase and antimitogillin antibodies in sera of patients suffering from invasive aspergillosis or aspergilloma, but not in control sera of healthy individuals. This suggests that chitosanase, like mitogillin, is expressed during infection and might therefore be of diagnostic relevance.
尽管致病微生物的分泌蛋白通常代表潜在的毒力因子，但迄今为止，关于烟曲霉分泌的蛋白质的信息仍然有限。因此，我们分析了在不同培养基中生长后的上清液蛋白。在无血清细胞培养基中，烟曲霉生长受限，未检测到蛋白质分泌，而在曲霉基本培养基（AMM）或更复杂的酵母葡萄糖培养基（YG）中生长后，可检测到明显的蛋白质图谱。在这些条件下分泌的三种主要蛋白质被鉴定为核糖毒素米托吉林、一种壳聚糖酶和曲霉胃蛋白酶i。米托吉林和壳聚糖酶在AMM中分泌，而曲霉胃蛋白酶i在YG中生长后尤为突出。当AMM培养物达到稳定期时，可检测到另外七种主要蛋白质。其中两种被鉴定为几丁质酶chiB1和一种β(1-3)内切葡聚糖酶。含有米托吉林和壳聚糖酶的条件培养基对A549和Vero细胞没有可检测到的细胞毒性作用。使用重组米托吉林和壳聚糖酶，我们在侵袭性曲霉病或曲霉肿患者的血清中检测到了抗壳聚糖酶和抗米托吉林抗体，但在健康个体的对照血清中未检测到。这表明壳聚糖酶与米托吉林一样，在感染期间表达，因此可能具有诊断意义。
