Pitarch Aida, Nombela César, Gil Concha
Department of Microbiology II, Faculty of Pharmacy, Complutense University of Madrid, Madrid, Spain.
Methods Mol Biol. 2009;470:369-411. doi: 10.1007/978-1-59745-204-5_26.
Candida albicans is a commensal inhabitant of the normal human microflora that can become pathogenic and invade almost all body sites and organs in response to both host-mediated and fungus-mediated mechanisms. Serologic responses to C. albicans that underlie its dichotomist relationship with the host (host-commensal and host-pathogen interactions) display a high degree of heterogeneity, resulting in distinct serum anti-Candida antibody signatures (molecular fingerprints of anti-Candida antibodies in serum) that can be used to discriminate commensal colonization from invasive disease. We describe the typical proteomic strategy to globally and integratively profile these host antibody responses and determine serum antibody signatures. This approach is based on the combination of classic immunoproteomics or serologic proteome analysis (two-dimensional electrophoresis followed by quantitative Western blotting and mass spectrometry) with data mining procedures. This global proteomic stratagem is a useful tool not only for obtaining an overview of different anti-Candida antibodies that are being elicited during the host-fungus interaction and, consequently, of the complex C. albicans immunome (the subset of the C. albicans proteome targeted by the immune system), but also for evaluating how this pathogen organism interacts with its host to trigger infection. In contrast with genomics and transcriptomics, this proteomic technology has the potential to detect antigenicity associated with posttranslational modification, subcellular localization, and other functional aspects that can be relevant in the host immune response. Furthermore, this strategy to define molecular fingerprints of serum anti-Candida antibodies may hopefully bring to light potential candidates for diagnosis, prognosis, risk stratification, clinical follow-up, therapeutic monitoring, and/or immunotherapy of candidiasis, especially of its life-threatening systemic forms.
白色念珠菌是人类正常微生物群的共生菌，在宿主介导和真菌介导的机制作用下，它可转变为致病菌并侵袭几乎所有身体部位和器官。白色念珠菌与宿主的二分关系（宿主-共生菌和宿主-病原体相互作用）背后的血清学反应表现出高度异质性，产生了独特的血清抗念珠菌抗体特征（血清中抗念珠菌抗体的分子指纹），可用于区分共生定植与侵袭性疾病。我们描述了一种典型的蛋白质组学策略，用于全面、综合地分析这些宿主抗体反应并确定血清抗体特征。该方法基于经典免疫蛋白质组学或血清蛋白质组分析（二维电泳，随后进行定量蛋白质印迹和质谱分析）与数据挖掘程序的结合。这种全局蛋白质组学策略不仅是一种有用的工具，可用于全面了解宿主-真菌相互作用过程中产生的不同抗念珠菌抗体，从而了解复杂的白色念珠菌免疫组（免疫系统靶向的白色念珠菌蛋白质组子集），还可用于评估这种病原体与宿主如何相互作用以引发感染。与基因组学和转录组学不同，这种蛋白质组学技术有潜力检测与翻译后修饰、亚细胞定位以及宿主免疫反应中可能相关的其他功能方面相关的抗原性。此外，这种定义血清抗念珠菌抗体分子指纹的策略有望揭示念珠菌病，尤其是其危及生命的全身形式的诊断、预后、风险分层、临床随访、治疗监测和/或免疫治疗的潜在候选物。
