Description of the interaction between Candida albicans and macrophages by mixed and quantitative proteome analysis without isolation

Abstract

Candida albicans is an opportunistic pathogen that causes fatal diseases in immunocompromised hosts. Host resistance against C. albicans relies on ingestion of the pathogen by macrophages. Analysis of the escaping behavior of C. albicans from macrophages is required to understand the onset of systemic candidiasis. In this study, native interactions of C. albicans with macrophages were investigated by proteome analysis using high efficiency of long monolithic silica capillary column. Using this system, we developed a method of “mixed and quantitative proteome analysis” in which C. albicans and macrophages were simultaneously analyzed by nanoLC–MS/MS without the need to isolate the two individual living cells. Two hundred twenty-seven proteins from C. albicans and five proteins from macrophages were identified as candidate interaction-specific molecules. C. albicans seemed to produce glucose through a β-oxidation pathway, a glyoxylate cycle, and gluconeogenesis for escape from macrophages. Up-regulation of stress-related and candidate pathogenic proteins in C. albicans indicated how C. albicans endured the harsh environment inside the macrophages. Down-regulation of apoptosis-associated protein NOA1- and chaperone HSPA1A-syntheses in macrophage indicated that C. albicans was able to escape from macrophages in part by suppressing the production of these macrophage proteins.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-015-0127-2) contains supplementary material, which is available to authorized users.

Dissecting Candida albicans Infection from the Perspective of C. albicans Virulence and Omics Approaches on Host–Pathogen Interaction: A Review

“…Kitahara et al analyzed the escaping behavior of C. albicans from macrophages by using mixed and quantitative proteome analysis in isolation [124]. The authors identified five proteins from macrophages and 237 proteins from C. albicans as candidate interaction-specific molecules.…”

“…Meanwhile, C. albicans -induced macrophage proteins were linked with a chaperone and apoptosis. The authors postulated that C. albicans can endure the harsh environment inside macrophages through upregulation of stress-related proteins, while the escape of C. albicans from macrophages could be associated with the production of glucose from a β-oxidation pathway and downregulation of chaperone HSPA1A- and apoptosis-associated protein, NOA1-syntheses, in macrophages [124]. In summary, proteome research on host–pathogen interaction with respect to Candida infection is still an emerging field.…”

Challenges and Strategies for Proteome Analysis of the Interaction of Human Pathogenic Fungi with Host Immune Cells

“…This approach has also been applied for the first report of a mixed and quantitative dual proteome analysis that has been recently published based on the interaction of C. albicans with murine macrophages. The authors identified 483 C. albicans and 1253 macrophage proteins in total, where of 227 C. albicans and five macrophage proteins of the co-culture experiment exhibited a differential expression compared to separate C. albicans and macrophage mono-culture controls [161]. Reporter ion quantification using the TMT labeling approach was applied to reveal relative quantitative changes on the proteome level [161].…”

“…The authors identified 483 C. albicans and 1253 macrophage proteins in total, where of 227 C. albicans and five macrophage proteins of the co-culture experiment exhibited a differential expression compared to separate C. albicans and macrophage mono-culture controls [161]. Reporter ion quantification using the TMT labeling approach was applied to reveal relative quantitative changes on the proteome level [161]. The protein median normalization approach cannot be recommended for all types of experimental setups and outcomes, in particular when a certain response on the actual protein level shows a significant but moderate differential regulation (e.g., 2.5-fold up) of numerous proteins in one direction and a drastic differential regulation (e.g., 25-fold down) of few proteins in the other direction.…”

The effect of fluconazole and amphotericin B on macrophage functions

“…Once activated, they act directly by destroying bacteria, parasites, viruses, fungi and tumor cells with reactive oxidants and hydrolytic enzymes, or indirectly by releasing specific mediators such as interleukin-1, tumor necrosis factor-alpha and others which can regulate other cells' activities. They are also responsible for processing the antigens and presenting the digested peptides to lymphocytes, while playing the leading role in damage repair (9)(10)(11) Original article and other compromises in host defense due to use of these therapies, it is important to reverse or at least lessen immunosuppresion in this patients (2,4). Fungal infections are treated with antifungal drugs.…”