Hydrolytic Gene Expression during Oroesophageal and Gastric Candidiasis in Immunocompetent and Immunodeficient Gnotobiotic Mice

Schofield, David A.; Westwater, Caroline; Warner, Thomas F.; Nicholas, Peter J.; Paulling, Emily E.; Balish, Edward

doi:10.1086/377182

Cited by 35 publications

(29 citation statements)

References 36 publications

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“…albicans SAP1-3 and SAP4-6 gene families are not pseudogenes that have subsisted to purifying selection and concerted evolution, but apparently have acquired new and specific functions in nitrogen metabolism, switching, and cell adherence (Cassone et al 2002, Felk et al 2002, Ripeau et al 2002, Schofield et al 2003. C. tropicalis has only one gene (SAPT4) related to the C. albicans SAP1-3 and SAP4-6 families.…”

Section: Discussionmentioning

confidence: 99%

Phylogeny and evolution of the aspartyl protease family from clinically relevant Candida species

Parra-Ortega

Cruz-Torres

Villa‐Tanaca

et al. 2009

Mem. Inst. Oswaldo Cruz

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Section: Discussionmentioning

confidence: 99%

Phylogeny and evolution of the aspartyl protease family from clinically relevant Candida species

Parra-Ortega

Cruz-Torres

Villa‐Tanaca

et al. 2009

Mem. Inst. Oswaldo Cruz

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“…Many authors have described the use of RT-PCR to study SAP gene expression in various disease models and human clinical specimens (Schaller et al, 1998(Schaller et al, , 2003Naglik et al, 1999Naglik et al, , 2003Ripeau et al, 2002;Schofield et al, 2003). Meaningful comparisons between the various SAP analyses and our ALS analysis are complicated by the varying detection limits for each assay.…”

Section: Discussionmentioning

confidence: 99%

“…For example, incorporation of radionucleotides into the RT-PCR products serves to increase limit of detection (Naglik et al, 1999;2003); limit of detection is also enhanced by Southern blotting of the RT-PCR products (Ripeau et al, 2002). Design of primers to amplify smaller and similarly sized PCR products increases detection limit and consistency among the results for each gene (Naglik et al, 1999;2003;Schofield et al, 2003). The overall body of work on RT-PCR analysis of SAP gene expression suggests differential expression across the family.…”

Section: Discussionmentioning

confidence: 99%

“…Such assays have been developed and used widely for analysis of gene expression for other C. albicans gene families (Schaller et al, 1998(Schaller et al, , 2003Hube et al, 2000;Ripeau et al, 2002;Naglik et al, 1999Naglik et al, , 2003Schofield et al, 2003). This paper describes development of an ALS gene RT-PCR assay and its application to cells from the reconstituted human buccal epithelium (RHE) disease model, which was originally introduced into the C. albicans literature by Schaller et al (1998).…”

Section: Introductionmentioning

confidence: 99%

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RT-PCR detection of Candida albicans ALS gene expression in the reconstituted human epithelium (RHE) model of oral candidiasis and in model biofilms

et al. 2004

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An RT-PCR assay was developed to analyse expression patterns of genes in the Candida albicans ALS (agglutinin-like sequence) family. Inoculation of a reconstituted human buccal epithelium (RHE) model of mucocutaneous candidiasis with strain SC5314 showed destruction of the epithelial layer by C. albicans and also formation of an upper fungal layer that had characteristics similar to a biofilm. RT-PCR analysis of total RNA samples extracted from C. albicans-inoculated buccal RHE showed that ALS1, ALS2, ALS3, ALS4, ALS5 and ALS9 were consistently detected over time as destruction of the RHE progressed. Detection of transcripts from ALS7, and particularly from ALS6, was more sporadic, but not associated with a strictly temporal pattern. The expression pattern of ALS genes in C. albicans cultures used to inoculate the RHE was similar to that observed in the RHE model, suggesting that contact of C. albicans with buccal RHE does little to alter ALS gene expression. RT-PCR analysis of RNA samples extracted from model denture and catheter biofilms showed similar gene expression patterns to the buccal RHE specimens. Results from the RT-PCR analysis of biofilm RNA specimens were consistent between various C. albicans strains during biofilm development and were comparable to gene expression patterns in planktonic cells. The RT-PCR assay described here will be useful for analysis of human clinical specimens and samples from other disease models. The method will provide further insight into the role of ALS genes and their encoded proteins in the diverse interactions between C. albicans and its host.

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“…Expression of SAP1, SAP2 and SAP4-SAP6 was detected by RT-PCR during invasion of parenchymal organs after intraperitoneal infection of mice (Felk et al, 2002). In a mouse model of oroesophageal and gastric candidiasis, all SAP genes were detectably expressed during gastric candidiasis, while SAP1 and SAP3 were only sporadically and weakly expressed at different oral sites of infection (Schofield et al, 2003). Several investigators have also used RT-PCR to analyse the expression of specific SAP genes during colonization and infection of humans.…”

Section: Introductionmentioning

confidence: 99%

Secreted aspartic proteases are not required for invasion of reconstituted human epithelia by Candida albicans

2008

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A well-known virulence attribute of the human-pathogenic yeast Candida albicans is the secretion of aspartic proteases (Saps), which may contribute to colonization and infection of different host niches by degrading tissue barriers, destroying host defence molecules, or digesting proteins for nutrient supply. The role of individual Sap isoenzymes, which are encoded by a large gene family, for the pathogenicity of C. albicans has been investigated by assessing the virulence of mutants lacking specific SAP genes and by studying the expression pattern of the SAP genes in various models of superficial and systemic infections. We used a recombination-based genetic reporter system to detect the induction of the SAP1-SAP6 genes during infection of reconstituted human vaginal epithelium. Only SAP5, but none of the other tested SAP genes, was detectably activated in this in vitro infection model. To directly address the importance of the SAP1-SAP6 genes for invasion of reconstituted human epithelia (RHE), we constructed a set of mutants of the wild-type C. albicans model strain SC5314 in which either single or multiple SAP genes were specifically deleted. Even mutants lacking all of the SAP1-SAP3 or the SAP4-SAP6 genes displayed the same capacity to invade and damage both oral and vaginal RHE as their wild-type parental strain, in contrast to a nonfilamentous efg1D mutant that was avirulent under these conditions. We therefore conclude from these results that the secreted aspartic proteases Sap1p-Sap6p are not required for invasion of RHE by C. albicans.

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Hydrolytic Gene Expression during Oroesophageal and Gastric Candidiasis in Immunocompetent and Immunodeficient Gnotobiotic Mice

Cited by 35 publications

References 36 publications

Phylogeny and evolution of the aspartyl protease family from clinically relevant Candida species

Phylogeny and evolution of the aspartyl protease family from clinically relevant Candida species

RT-PCR detection of Candida albicans ALS gene expression in the reconstituted human epithelium (RHE) model of oral candidiasis and in model biofilms

Secreted aspartic proteases are not required for invasion of reconstituted human epithelia by Candida albicans

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