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Gene profiling and signaling pathways of Candida albicans keratitis.

Yuan X, Mitchell BM, Wilhelmus KR - Mol. Vis. (2008)

Bottom Line: Selected genes encoding interleukins (IL), chemokine ligands, and other cytokines were confirmed by quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR).Pathways having more than 20% of their genes significantly upregulated signaled leukocyte extravasation, increased interleukin production, and affected toll-like receptors.Upregulated transcript levels for IL-1beta and IL-6 were confirmed by real-time RT-PCR.

View Article: PubMed Central - PubMed

Affiliation: Sid W Richardson Ocular Microbiology Laboratory, Cullen Eye Institute, Baylor College of Medicine, Houston, TX, USA.

ABSTRACT

Purpose: To compare the global gene expression patterns in uninfected and fungus-infected mouse corneas at the onset of Candida albicans keratitis.

Methods: Fungal keratitis was generated by scarifying the corneal epithelium of BALB/c mice followed by topical inoculation with Candida albicans. Corneal infection was allowed to progress for one day, and total RNA was then extracted from excised corneas. Microarray was performed to detect 45,102 murine genes and processed to identify genetic regulation of signaling pathways. Selected genes encoding interleukins (IL), chemokine ligands, and other cytokines were confirmed by quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR).

Results: Compared to mock-inoculated control eyes, genetic microarray analysis of Candida albicans keratitis showed that 3,977 genes (8.8%) changed at least twofold and 1,672 genes (3.7%) changed at least fourfold. Hierarchical clustering identified that upregulated genes affected immune and inflammatory responses, intercellular signaling, and cellular proliferation. Pathways having more than 20% of their genes significantly upregulated signaled leukocyte extravasation, increased interleukin production, and affected toll-like receptors. Upregulated transcript levels for IL-1beta and IL-6 were confirmed by real-time RT-PCR.

Conclusions: Host gene expression during the initial stage of Candida albicans keratitis involves pathways contributing to acute inflammation mediated by interleukins and other signals of leukocyte recruitment. This murine study confirms the involvement of innate immunity in the cornea during the initiation of Candida albicans keratitis.

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Related in: MedlinePlus

Differential upregulation and down-regulation of corneal genes. Microarray results compare gene expression levels in mouse corneas from Candida albicans-infected compared to corneas from mock-infected controls. The results were assigned to general categories based upon the known functions of the gene products. The number of genes expressing a twofold or greater significant difference (p≤0.05) is plotted for the listed categories. Red bars represent the number of genes significantly down-regulated, and black bars represent the number of genes significantly upregulated.
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f2: Differential upregulation and down-regulation of corneal genes. Microarray results compare gene expression levels in mouse corneas from Candida albicans-infected compared to corneas from mock-infected controls. The results were assigned to general categories based upon the known functions of the gene products. The number of genes expressing a twofold or greater significant difference (p≤0.05) is plotted for the listed categories. Red bars represent the number of genes significantly down-regulated, and black bars represent the number of genes significantly upregulated.

Mentions: Gene expression levels were assigned to general categories based upon known functions of gene products and the number of genes expressing greater than or equal to twofold significant difference (p≤0.05). While functional categories included some down-regulated genes, the number of upregulated genes was consistently more prevalent (Figure 2). Canonical analysis showed that the ratio of significantly upregulated genes within the pathways ranged from 0.13 to 0.34 (Table 4). Several signaling pathways included genes that were upregulated fourfold or more (Appendix 1). Similar results were found for selected genes detected by microarray and real-time reverse transcriptase (RT)–PCR analysis (Table 5), although CCR3 expression differed between the infected and mock-infected corneas.


Gene profiling and signaling pathways of Candida albicans keratitis.

Yuan X, Mitchell BM, Wilhelmus KR - Mol. Vis. (2008)

Differential upregulation and down-regulation of corneal genes. Microarray results compare gene expression levels in mouse corneas from Candida albicans-infected compared to corneas from mock-infected controls. The results were assigned to general categories based upon the known functions of the gene products. The number of genes expressing a twofold or greater significant difference (p≤0.05) is plotted for the listed categories. Red bars represent the number of genes significantly down-regulated, and black bars represent the number of genes significantly upregulated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2562425&req=5

f2: Differential upregulation and down-regulation of corneal genes. Microarray results compare gene expression levels in mouse corneas from Candida albicans-infected compared to corneas from mock-infected controls. The results were assigned to general categories based upon the known functions of the gene products. The number of genes expressing a twofold or greater significant difference (p≤0.05) is plotted for the listed categories. Red bars represent the number of genes significantly down-regulated, and black bars represent the number of genes significantly upregulated.
Mentions: Gene expression levels were assigned to general categories based upon known functions of gene products and the number of genes expressing greater than or equal to twofold significant difference (p≤0.05). While functional categories included some down-regulated genes, the number of upregulated genes was consistently more prevalent (Figure 2). Canonical analysis showed that the ratio of significantly upregulated genes within the pathways ranged from 0.13 to 0.34 (Table 4). Several signaling pathways included genes that were upregulated fourfold or more (Appendix 1). Similar results were found for selected genes detected by microarray and real-time reverse transcriptase (RT)–PCR analysis (Table 5), although CCR3 expression differed between the infected and mock-infected corneas.

Bottom Line: Selected genes encoding interleukins (IL), chemokine ligands, and other cytokines were confirmed by quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR).Pathways having more than 20% of their genes significantly upregulated signaled leukocyte extravasation, increased interleukin production, and affected toll-like receptors.Upregulated transcript levels for IL-1beta and IL-6 were confirmed by real-time RT-PCR.

View Article: PubMed Central - PubMed

Affiliation: Sid W Richardson Ocular Microbiology Laboratory, Cullen Eye Institute, Baylor College of Medicine, Houston, TX, USA.

ABSTRACT

Purpose: To compare the global gene expression patterns in uninfected and fungus-infected mouse corneas at the onset of Candida albicans keratitis.

Methods: Fungal keratitis was generated by scarifying the corneal epithelium of BALB/c mice followed by topical inoculation with Candida albicans. Corneal infection was allowed to progress for one day, and total RNA was then extracted from excised corneas. Microarray was performed to detect 45,102 murine genes and processed to identify genetic regulation of signaling pathways. Selected genes encoding interleukins (IL), chemokine ligands, and other cytokines were confirmed by quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR).

Results: Compared to mock-inoculated control eyes, genetic microarray analysis of Candida albicans keratitis showed that 3,977 genes (8.8%) changed at least twofold and 1,672 genes (3.7%) changed at least fourfold. Hierarchical clustering identified that upregulated genes affected immune and inflammatory responses, intercellular signaling, and cellular proliferation. Pathways having more than 20% of their genes significantly upregulated signaled leukocyte extravasation, increased interleukin production, and affected toll-like receptors. Upregulated transcript levels for IL-1beta and IL-6 were confirmed by real-time RT-PCR.

Conclusions: Host gene expression during the initial stage of Candida albicans keratitis involves pathways contributing to acute inflammation mediated by interleukins and other signals of leukocyte recruitment. This murine study confirms the involvement of innate immunity in the cornea during the initiation of Candida albicans keratitis.

Show MeSH
Related in: MedlinePlus