Coelho Paulo S R, Im Hogune, Clemons Karl V, Snyder Michael P, Stevens David A
Division of Infectious Diseases and Geographic Medicine, Stanford University , 300 Pasteur Drive, Lane L-134, Stanford, California 94305-5107, United States.
California Institute for Medical Research , 2260 Clove Drive, San Jose, California 95128, United States.
J Proteome Res. 2015 Sep 4;14(9):3924-31. doi: 10.1021/acs.jproteome.5b00365. Epub 2015 Aug 21.
We profiled the global immunoglobulin response against fungal infection by using yeast protein microarrays. Groups of CD-1 mice were infected systemically with human fungal pathogens (Coccidioides posadasii, Candida albicans, or Paracoccidioides brasiliensis) or inoculated with PBS as a control. Another group was inoculated with heat-killed yeast (HKY) of Saccharomyces cerevisiae. After 30 days, serum from mice in the groups were collected and used to probe S. cerevisiae protein microarrays containing 4800 full-length glutathione S-transferase (GST)-fusion proteins. Antimouse IgG conjugated with Alexafluor 555 and anti-GST antibody conjugated with Alexafluor 647 were used to detect antibody-antigen interactions and the presence of GST-fusion proteins, respectively. Serum after infection with C. albicans reacted with 121 proteins: C. posadasii, 81; P. brasiliensis, 67; and after HKY, 63 proteins on the yeast protein microarray, respectively. We identified a set of 16 antigenic proteins that were shared across the three fungal pathogens. These include retrotransposon capsid proteins, heat shock proteins, and mitochondrial proteins. Five of these proteins were identified in our previous study of fungal cell wall by mass spectrometry (Ann. N. Y. Acad. Sci. 2012, 1273, 44-51). The results obtained give a comprehensive view of the immunological responses to fungal infections at the proteomic level. They also offer insight into immunoreactive protein commonality among several fungal pathogens and provide a basis for a panfungal vaccine.
我们通过使用酵母蛋白微阵列分析了针对真菌感染的全球免疫球蛋白反应。将多组CD-1小鼠全身感染人类真菌病原体（波萨达斯球孢子菌、白色念珠菌或巴西副球孢子菌），或接种PBS作为对照。另一组接种酿酒酵母的热灭活酵母（HKY）。30天后，收集各组小鼠的血清，用于探测包含4800种全长谷胱甘肽S-转移酶（GST）融合蛋白的酿酒酵母蛋白微阵列。分别使用与Alexafluor 555偶联的抗小鼠IgG和与Alexafluor 647偶联的抗GST抗体来检测抗体-抗原相互作用以及GST融合蛋白的存在。白色念珠菌感染后的血清与121种蛋白质发生反应：波萨达斯球孢子菌感染后的血清与81种蛋白质发生反应；巴西副球孢子菌感染后的血清与67种蛋白质发生反应；HKY接种后的血清与酵母蛋白微阵列上的63种蛋白质发生反应。我们鉴定出一组在三种真菌病原体中都存在的16种抗原性蛋白质。这些蛋白质包括逆转座子衣壳蛋白、热休克蛋白和线粒体蛋白。其中有5种蛋白质在我们之前通过质谱对真菌细胞壁的研究中已被鉴定（《纽约科学院学报》，2012年，第1273卷，第44 - 51页）。所获得的结果在蛋白质组水平上全面展示了对真菌感染的免疫反应。它们还为几种真菌病原体之间的免疫反应性蛋白质共性提供了见解，并为泛真菌疫苗提供了基础。
