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Identification of Candida albicans genes expressed during oropharyngeal candidiasis

, : Identification of Candida albicans genes expressed during oropharyngeal candidiasis. Abstracts of the Interscience Conference on Antimicrobial Agents & Chemotherapy 41: 377

Candida albicans regulates expression of multiple genes during infection. Previous studies of C. albicans gene expression in vivo have extrapolated findings from in vitro systems or animal models, which might not be relevant to human infections. In the present study, we used In Vivo Induced Antigen Technology (IVIAT) to identify genes expressed by C. albicans during oropharyngeal candidiasis (OPC) among HIV-infected patients. Pooled sera from 5 patients with OPC were adsorbed with lab-grown whole cells and cell lysates of 3 C. albicans clinical strains. A C. albicans expression library was created by ligating genomic fragments from clinical isolates into pET30 vector. The adsorbed sera were used to screen the library by colony blot, plasmid inserts from reactive colonies were sequenced, and C. albicans ORFs identified. Thus far, we have screened over 40,000 colonies, and identified 34 ORFs that encode antigens that are repeatedly reactive with sera. To date, the reactivity of 14 of these ORFs have been confirmed by Western analysis. We next verified that expression of each of the 14 ORFs was upregulated in vivo by quantitative RT-PCR using template RNA obtained from patients' OPC samples versus RNA from in vitro grown cells, and standardized to expression of EBF-1. Eight of the in vivo induced ORFs represent previously identified C. albicans genes, while 6 represent previously unidentified genes with homology to S. cerevisiae genes. The genes fall into 5 distinct classes: 1) adaptive responses to stress (4 genes), including DNA repair and tolerance to heat, acidity, and elevated glucose concentrations; 2) regulatory functions (4), including regulation of yeast-to-hyphal morphogenesis (3) and cell division (1); 3) energy utitlization and acquisition of amino acids or nucleotides (2); 4) cell surface structure and phospholipid metabolism (2); 5) genes of unknown function (2). Four of our induced genes represent known virulence factors for C. albicans.


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