Immunoproteomic analysis of antibody responses to extracellular proteins of Candida albicans revealed the importance of glycosylation for antigen recognition.

Luo T, Krüger T, Knüpfer U, Kasper L, Wielsch N, Hube B, Kortgen A, Bauer M, Giamarellos-Bourboulis EJ, Dimopoulos G, Brakhage AA, Kniemeyer O (2016) Immunoproteomic analysis of antibody responses to extracellular proteins of Candida albicans revealed the importance of glycosylation for antigen recognition. J Proteome Res 15(8), 2394-2406.

Abstract

During infection, the human pathogenic fungus Candida albicans undergoes a yeast-to-hypha transition, secretes numerous proteins for invasion of host tissues and modulates the host's immune response. Little is known about the interplay of C. albicans secreted proteins and the host adaptive immune system. Here, we applied a combined 2D gel- and LC-MS/MS-based approach for the characterization of C. albicans extracellular proteins during the yeast-to-hypha transition, which led to a comprehensive C. albicans secretome map. The serological responses to C. albicans extracellular proteins were investigated by a 2D-immunoblotting approach combined with MS for protein identification. Based on the screening of sera from candidemia and three groups of non-candidemia patients, a core set of 19 immunodominant antibodies against secreted proteins of C. albicans was identified, seven of which represent potential diagnostic markers for candidemia (Xog1, Lip4, Asc1, Met6, Tsa1, Tpi1 and Prx1). Intriguingly, some secreted, strongly glycosylated protein antigens showed high cross-reactivity with sera from non-candidemia control groups. Enzymatic deglycosylation of proteins secreted from hyphae significantly impair sera antibody recognition. Furthermore, deglycosylation of the recombinantly produced, secreted aspartyl protease Sap6 confirmed a significant contribution of glycan epitopes to the recognition of Sap6 by antibodies in patients´ sera.

Leibniz-HKI-Authors

Axel A. Brakhage
Bernhard Hube
Lydia Kasper
Olaf Kniemeyer
Uwe Knüpfer
Thomas Krüger
Ting Luo

Identifier

doi: 10.1021/acs.jproteome.5b01065

PMID: 27386892