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Polymicrobial Oral Infection with Four Periodontal Bacteria Orchestrates a Distinct Inflammatory Response and Atherosclerosis in ApoE Mice.

Chukkapalli SS, Velsko IM, Rivera-Kweh MF, Zheng D, Lucas AR, Kesavalu L - PLoS ONE (2015)

Bottom Line: Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis.Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES.In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, College of Dentistry, University of Florida, Gainesville, Florida, United States of America.

ABSTRACT
Periodontal disease (PD) develops from a synergy of complex subgingival oral microbiome, and is linked to systemic inflammatory atherosclerotic vascular disease (ASVD). To investigate how a polybacterial microbiome infection influences atherosclerotic plaque progression, we infected the oral cavity of ApoE mice with a polybacterial consortium of 4 well-characterized periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, Tannerealla forsythia and Fusobacterium nucleatum, that have been identified in human atherosclerotic plaque by DNA screening. We assessed periodontal disease characteristics, hematogenous dissemination of bacteria, peripheral T cell response, serum inflammatory cytokines, atherosclerosis risk factors, atherosclerotic plaque development, and alteration of aortic gene expression. Polybacterial infections have established gingival colonization in ApoE hyperlipidemic mice and displayed invasive characteristics with hematogenous dissemination into cardiovascular tissues such as the heart and aorta. Polybacterial infection induced significantly higher levels of serum risk factors oxidized LDL (p < 0.05), nitric oxide (p < 0.01), altered lipid profiles (cholesterol, triglycerides, Chylomicrons, VLDL) (p < 0.05) as well as accelerated aortic plaque formation in ApoE mice (p < 0.05). Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis. Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES. In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules. This study demonstrates unique differences in the host immune response to a polybacterial periodontal infection with atherosclerotic lesion progression in a mouse model.

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Polybacterial infection elicits a distinct splenic T cell response.Representative images of the gating scheme are shown in panels 2 and 3, with panel 2 showing the lymphocyte gate, panel 3 showing the CD3+ CD4+ lymphocyte gate, and panel 4 showing the histogram of CD3+ CD4+ Receptor+ lymphocytes. (A). CD3+ CD4+ IFNγR+ cells indicate Th1 response. (B). CD3+ CD4+ IL4R+ cells indicate Th2 response. (C). CD3+ CD4+ IL17R+ cells indicate Th17 response. Poly—polybacterial infection. ** p < 0.01.
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pone.0143291.g002: Polybacterial infection elicits a distinct splenic T cell response.Representative images of the gating scheme are shown in panels 2 and 3, with panel 2 showing the lymphocyte gate, panel 3 showing the CD3+ CD4+ lymphocyte gate, and panel 4 showing the histogram of CD3+ CD4+ Receptor+ lymphocytes. (A). CD3+ CD4+ IFNγR+ cells indicate Th1 response. (B). CD3+ CD4+ IL4R+ cells indicate Th2 response. (C). CD3+ CD4+ IL17R+ cells indicate Th17 response. Poly—polybacterial infection. ** p < 0.01.

Mentions: In order to evaluate the T cell responses to polybacterial infection, the splenic T cell profile was assessed at 12 weeks of infection by flow cytometry. Specifically, the polarization of the T cell pool to that of a Th1, Th2 or Th17 phenotype was evaluated based on the percentage of T cells expressing IFNγR, IL4R or IL17R, respectively. The percentage of Th1 cells, characterized by expression of IFNγR, was not significantly different between the polybacterial-infected mice and sham-infected mice (Fig 2A). Polybacterial-infected mice had significantly (p < 0.01) elevated levels of IL4R-expressing T cells relative to sham-infected mice (Fig 2B), indicating a strong Th2 response, which aligns with the significant serum IgG and IgM antibody response. IL17R-expressing Th17 cell numbers were not significantly different between polybacterial-infected and sham-infected mice (Fig 2C). Further, we determined serum cytokine alteration by an inflammatory cytokine array to assess how chronic polybacterial infection altered systemic cytokine levels at 12 (n = 6) and 24 weeks (n = 6) of polybacterial infection. At 12 weeks, infected mice demonstrated decreased levels (> 2-fold) of three cytokines, Fas ligand, Interleukin (IL) IL-13, and SDF-1 (Stromal cell-derived factor 1), with increased levels of the chemokine RANTES (Regulated upon activation, Normal T cell expressed and secreted) relative to sham-infected mice (Table 3), indicating an altered inflammatory cytokine response to infection. By 24 weeks of infection, two cytokines involved in attracting and promoting proliferation of innate immune cells, KC (Keratinocyte Chemoattractant) and MCSF (Macrophage colony stimulating factor), were detected at lower levels in infected mice than sham-infected mice (Table 3). Eight cytokines were detected at levels greater than 2-fold higher in infected mice when compared to controls, including the strongly-up-regulated Th2-promoting cytokines IL-3, IL-4, and IL-13, which corresponds to the markedly increased serum antibody response and splenic T cell polarization. Additionally the T cell chemoattractants lymphotactin and RANTES were up-regulated, supporting T cell recruitment in response to the polybacterial infection. Elevated interferon gamma (IFNγ) and IL-1β also indicated infection-enhanced inflammation.


Polymicrobial Oral Infection with Four Periodontal Bacteria Orchestrates a Distinct Inflammatory Response and Atherosclerosis in ApoE Mice.

Chukkapalli SS, Velsko IM, Rivera-Kweh MF, Zheng D, Lucas AR, Kesavalu L - PLoS ONE (2015)

Polybacterial infection elicits a distinct splenic T cell response.Representative images of the gating scheme are shown in panels 2 and 3, with panel 2 showing the lymphocyte gate, panel 3 showing the CD3+ CD4+ lymphocyte gate, and panel 4 showing the histogram of CD3+ CD4+ Receptor+ lymphocytes. (A). CD3+ CD4+ IFNγR+ cells indicate Th1 response. (B). CD3+ CD4+ IL4R+ cells indicate Th2 response. (C). CD3+ CD4+ IL17R+ cells indicate Th17 response. Poly—polybacterial infection. ** p < 0.01.
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Related In: Results  -  Collection

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pone.0143291.g002: Polybacterial infection elicits a distinct splenic T cell response.Representative images of the gating scheme are shown in panels 2 and 3, with panel 2 showing the lymphocyte gate, panel 3 showing the CD3+ CD4+ lymphocyte gate, and panel 4 showing the histogram of CD3+ CD4+ Receptor+ lymphocytes. (A). CD3+ CD4+ IFNγR+ cells indicate Th1 response. (B). CD3+ CD4+ IL4R+ cells indicate Th2 response. (C). CD3+ CD4+ IL17R+ cells indicate Th17 response. Poly—polybacterial infection. ** p < 0.01.
Mentions: In order to evaluate the T cell responses to polybacterial infection, the splenic T cell profile was assessed at 12 weeks of infection by flow cytometry. Specifically, the polarization of the T cell pool to that of a Th1, Th2 or Th17 phenotype was evaluated based on the percentage of T cells expressing IFNγR, IL4R or IL17R, respectively. The percentage of Th1 cells, characterized by expression of IFNγR, was not significantly different between the polybacterial-infected mice and sham-infected mice (Fig 2A). Polybacterial-infected mice had significantly (p < 0.01) elevated levels of IL4R-expressing T cells relative to sham-infected mice (Fig 2B), indicating a strong Th2 response, which aligns with the significant serum IgG and IgM antibody response. IL17R-expressing Th17 cell numbers were not significantly different between polybacterial-infected and sham-infected mice (Fig 2C). Further, we determined serum cytokine alteration by an inflammatory cytokine array to assess how chronic polybacterial infection altered systemic cytokine levels at 12 (n = 6) and 24 weeks (n = 6) of polybacterial infection. At 12 weeks, infected mice demonstrated decreased levels (> 2-fold) of three cytokines, Fas ligand, Interleukin (IL) IL-13, and SDF-1 (Stromal cell-derived factor 1), with increased levels of the chemokine RANTES (Regulated upon activation, Normal T cell expressed and secreted) relative to sham-infected mice (Table 3), indicating an altered inflammatory cytokine response to infection. By 24 weeks of infection, two cytokines involved in attracting and promoting proliferation of innate immune cells, KC (Keratinocyte Chemoattractant) and MCSF (Macrophage colony stimulating factor), were detected at lower levels in infected mice than sham-infected mice (Table 3). Eight cytokines were detected at levels greater than 2-fold higher in infected mice when compared to controls, including the strongly-up-regulated Th2-promoting cytokines IL-3, IL-4, and IL-13, which corresponds to the markedly increased serum antibody response and splenic T cell polarization. Additionally the T cell chemoattractants lymphotactin and RANTES were up-regulated, supporting T cell recruitment in response to the polybacterial infection. Elevated interferon gamma (IFNγ) and IL-1β also indicated infection-enhanced inflammation.

Bottom Line: Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis.Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES.In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, College of Dentistry, University of Florida, Gainesville, Florida, United States of America.

ABSTRACT
Periodontal disease (PD) develops from a synergy of complex subgingival oral microbiome, and is linked to systemic inflammatory atherosclerotic vascular disease (ASVD). To investigate how a polybacterial microbiome infection influences atherosclerotic plaque progression, we infected the oral cavity of ApoE mice with a polybacterial consortium of 4 well-characterized periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, Tannerealla forsythia and Fusobacterium nucleatum, that have been identified in human atherosclerotic plaque by DNA screening. We assessed periodontal disease characteristics, hematogenous dissemination of bacteria, peripheral T cell response, serum inflammatory cytokines, atherosclerosis risk factors, atherosclerotic plaque development, and alteration of aortic gene expression. Polybacterial infections have established gingival colonization in ApoE hyperlipidemic mice and displayed invasive characteristics with hematogenous dissemination into cardiovascular tissues such as the heart and aorta. Polybacterial infection induced significantly higher levels of serum risk factors oxidized LDL (p < 0.05), nitric oxide (p < 0.01), altered lipid profiles (cholesterol, triglycerides, Chylomicrons, VLDL) (p < 0.05) as well as accelerated aortic plaque formation in ApoE mice (p < 0.05). Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis. Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES. In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules. This study demonstrates unique differences in the host immune response to a polybacterial periodontal infection with atherosclerotic lesion progression in a mouse model.

Show MeSH
Related in: MedlinePlus