Evaluating Common Humoral Responses against Fungal Infections with Yeast Protein Microarrays.
Evaluating Common Humoral Responses against Fungal Infections with Yeast Protein Microarrays.
复制标题
使用酵母蛋白微阵列评估针对真菌感染的常见体液反应。
DOI：
10.1021/acs.jproteome.5b00365
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发表时间：
2015
影响因子：
4.4
通讯作者：
Stevens,DavidA
中科院分区：
文献类型：
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作者：
Coelho,PauloSR;Im,Hogune;Clemons,KarlV;Snyder,MichaelP;Stevens,DavidA
我们通过使用酵母蛋白微阵列分析了针对真菌感染的整体免疫球蛋白反应。 CD-1 组小鼠全身感染人类真菌病原体（波萨达球孢子菌、白色念珠菌或巴西副球孢子菌），或接种 PBS 作为对照。另一组接种了酿酒酵母的热灭活酵母(HKY)。 30天后，收集各组小鼠的血清并用于探测含有4800个全长谷胱甘肽S-转移酶(GST)融合蛋白的酿酒酵母蛋白微阵列。与 Alexa Fluor 555 缀合的抗小鼠 IgG 和与 Alexa Fluor 647 缀合的抗 GST 抗体分别用于检测抗体-抗原相互作用和 GST 融合蛋白的存在。感染后的血清C.白色念珠菌与 121 种蛋白质发生反应：C.波萨达西，81；P。巴西人，67； HKY之后，酵母蛋白微阵列上分别有63个蛋白。我们鉴定了三种真菌病原体共有的一组 16 种抗原蛋白。这些包括逆转录转座子衣壳蛋白、热休克蛋白和线粒体蛋白。我们之前通过质谱法对真菌细胞壁进行的研究中鉴定出了其中的五种蛋白质 (Ann. N. Y. Acad. Sci.2012,1273, 44–51)。获得的结果在蛋白质组水平上全面了解了真菌感染的免疫反应。它们还提供了对几种真菌病原体之间免疫反应蛋白共性的深入了解，并为全真菌疫苗提供了基础。
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, orParacoccidioides brasiliensis) or inoculated with PBS as a control. Another group was inoculated with heat-killed yeast (HKY) ofSaccharomyces cerevisiae. After 30 days, serum from mice in the groups were collected and used to probeS.cerevisiaeprotein 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 withC. albicansreacted 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.