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. 2007 May;75(5):2540-7.
doi: 10.1128/IAI.01957-06. Epub 2007 Feb 16.

Gingival epithelial cell transcriptional responses to commensal and opportunistic oral microbial species (V体育平台登录)

Yoshiaki Hasegawa  1 , Jeffrey J Mans, Song Mao, M Cecilia Lopez, Henry V Baker, Martin Handfield, Richard J Lamont
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Gingival epithelial cell transcriptional responses to commensal and opportunistic oral microbial species (V体育官网)

Yoshiaki Hasegawa et al. Infect Immun. 2007 May.

Abstract (V体育平台登录)

Transcriptional profiling and ontology tools were utilized to define the biological pathways of gingival epithelial cells modulated by coculture with the oral commensal Streptococcus gordonii and the opportunistic commensal Fusobacterium nucleatum. Overall, F. nucleatum and S. gordonii perturbed the gingival epithelial cell transcriptome much less significantly than the oral pathogens Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans perturbed the transcriptome, indicating that there was a greater degree of host adaptation by the commensal species (M. Handfield, J. J. Mans, G. Zheng, M. C. Lopez, S. Mao, A. Progulske-Fox, G. Narasimhan, H. V. Baker, and R. J. Lamont, Cell. Microbiol. 7:811-823, 2005) VSports手机版. The biological pathways significantly impacted by F. nucleatum and S. gordonii included the mitogen-activated protein kinase (MAPK) and Toll-like receptor signaling pathways. Differential regulation of GADD45 and DUSP4, key components of the MAPK pathway, was confirmed at the protein level by Western blotting. Modulation of the MAPK pathway is likely to affect host cell proliferation and differentiation. In addition, both the MAPK and Toll-like receptor pathways ultimately converge on cytokine gene expression. An enzyme-linked immunosorbent assay of secreted interleukin-6 (IL-6) and IL-8 demonstrated that F. nucleatum induced production of these cytokines, whereas S. gordonii inhibited secretion from the epithelial cells. Stimulation of secretion of proinflammatory cytokines from epithelial cells may reflect the invasive phenotype of F. nucleatum and contribute to the greater pathogenic potential of F. nucleatum than of S. gordonii. .

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Figures

FIG. 1.
FIG. 1.
Hierarchical clustering of variance-normalized gene expression data. The expression pattern of the cRNAs analyzed by using microarrays was determined by a supervised analysis of the variance-normalized data set of differentially expressed genes (P < 0.001, BRB ArrayTools) with the algorithm Cluster and was displayed with Treeview. Each row represents an individual gene element on the array, and each column represents the expression states of cRNAs for the challenge condition indicated. Each expression data point represents the relative fluorescence intensity of the cRNA from F. nucleatum-infected cells (columns Fn R1 to Fn R4) or S. gordonii-infected cells (columns Sg R1 to Sg R4) to the fluorescence intensity of the cRNA from uninfected cells (columns CTRL R1 to CTRL R4). The distance matrix used to determine the relatedness of samples through gene expression space was 1 − Pearson's correlation coefficient. The cluster is subdivided into three groups consisting of genes that were repressed (green), genes that were induced (red), and genes whose expression did not change (black). The variation in gene expression for a given gene is expressed as the distance from the mean observation for that gene according to the color scale below the heat map.
FIG. 2.
FIG. 2.
Western immunoblots of HIGK infected with F. nucleatum (Fn) or S. gordonii (Sg) and uninfected controls (CTRL) for 1, 2, or 6 h. The blots were probed with antibodies to GADD45α (upper panel), GADD45β (middle panel), and DUSP4 (lower panel) and then stripped and reprobed with antibodies to β-actin. The graphs show the results of densitometric analyses of the ratio of test protein band intensity to β-actin band intensity.
FIG. 3.
FIG. 3.
MAPK-related pathways containing F. nucleatum and S. gordonii differentially regulated genes at P < 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature (see text for details). Red indicates up-regulation, green indicates down-regulation, and gray indicates no change in expression. +P indicates phosphorylation, and −P indicates dephosphorylation. An arrow indicates a molecular interaction resulting in activation, and a line without an arrowhead indicates a molecular interaction resulting in inhibition. e, expression.
FIG. 4.
FIG. 4.
Enzyme-linked immunosorbent assay of IL-6 (A) and IL-8 (B) accumulation in HIGK supernatants following coculture with F. nucleatum or S. gordonii for 2, 4, 6, or 8 h. Ctrl, uninfected control. The error bars indicate standard deviations (n = 3). One asterisk, P < 0.05; two asterisks, P < 0.001 (as determined by a t test).
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References

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