Impact of commensal flora on periodontal immune response to lipopolysaccharide

Daiki Fukuhara, Koichiro Irie, Yoko Uchida, Kota Kataoka, Kentaro Akiyama, Daisuke Ekuni, Takaaki Tomofuji, Manabu Morita

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background: Commensal flora are involved in the appropriate development of the mature immune system. However, it is unclear how commensal flora contribute to immune responses against periodontal pathogens, including the response to lipopolysaccharide (LPS). The purpose of this study was to evaluate the expression of immune responses after topical application of LPS in germ-free (GF) and specific-pathogen-free (SPF) mice. Methods: GF and SPF mice at 8 weeks of age were randomly divided into four groups each: a baseline group (n = 4/group) and three experimental groups (n = 6/group). Experimental groups received topical application of Porphyromonas gingivalis LPS (10 μg/μL) into the palatal gingival sulcus. Sampling was performed before LPS application (baseline) and at 3, 24, or 72 hours after LPS application. The numbers of neutrophils, CD4+, and CD8+ T cells in periodontal tissue were evaluated by immunohistochemistry. Expression of genes encoding cytokines, chemokines, and a transcription factor was determined by real-time PCR. Results: SPF mice, but not GF mice, showed an increased number of CD4+ T cells in the periodontal tissue at 3 hours after LPS application, compared with the number at baseline (p < 0.05). Gene expressions of tumor necrosis factor-α (Tnf-α) and forkhead box protein p3 (Foxp3) was also significantly higher in the SPF mice than in the GF mice at 3 hours after LPS application (p < 0.05). The number of neutrophils peaked at 24 hours in both GF and SPF mice. Conclusions: LPS-exposed SPF mice exhibited increases in the number of CD4+ T cells and in Tnf-α and Foxp3 gene expression in periodontal tissue compared with LPS-exposed GF mice.

Original languageEnglish
Pages (from-to)1213-1220
Number of pages8
JournalJournal of Periodontology
Volume89
Issue number10
DOIs
Publication statusPublished - Jan 1 2018

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Lipopolysaccharides
Specific Pathogen-Free Organisms
T-Lymphocytes
Gene Expression
Neutrophils
Tumor Necrosis Factor-alpha
Forkhead Transcription Factors
Porphyromonas gingivalis
Chemokines
Real-Time Polymerase Chain Reaction
Immune System
Transcription Factors
Immunohistochemistry
Cytokines

Keywords

  • Animal model
  • Immune system
  • Lipopolysaccharide
  • Microbiota

ASJC Scopus subject areas

  • Periodontics

Cite this

Impact of commensal flora on periodontal immune response to lipopolysaccharide. / Fukuhara, Daiki; Irie, Koichiro; Uchida, Yoko; Kataoka, Kota; Akiyama, Kentaro; Ekuni, Daisuke; Tomofuji, Takaaki; Morita, Manabu.

In: Journal of Periodontology, Vol. 89, No. 10, 01.01.2018, p. 1213-1220.

Research output: Contribution to journalArticle

Fukuhara, Daiki ; Irie, Koichiro ; Uchida, Yoko ; Kataoka, Kota ; Akiyama, Kentaro ; Ekuni, Daisuke ; Tomofuji, Takaaki ; Morita, Manabu. / Impact of commensal flora on periodontal immune response to lipopolysaccharide. In: Journal of Periodontology. 2018 ; Vol. 89, No. 10. pp. 1213-1220.
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abstract = "Background: Commensal flora are involved in the appropriate development of the mature immune system. However, it is unclear how commensal flora contribute to immune responses against periodontal pathogens, including the response to lipopolysaccharide (LPS). The purpose of this study was to evaluate the expression of immune responses after topical application of LPS in germ-free (GF) and specific-pathogen-free (SPF) mice. Methods: GF and SPF mice at 8 weeks of age were randomly divided into four groups each: a baseline group (n = 4/group) and three experimental groups (n = 6/group). Experimental groups received topical application of Porphyromonas gingivalis LPS (10 μg/μL) into the palatal gingival sulcus. Sampling was performed before LPS application (baseline) and at 3, 24, or 72 hours after LPS application. The numbers of neutrophils, CD4+, and CD8+ T cells in periodontal tissue were evaluated by immunohistochemistry. Expression of genes encoding cytokines, chemokines, and a transcription factor was determined by real-time PCR. Results: SPF mice, but not GF mice, showed an increased number of CD4+ T cells in the periodontal tissue at 3 hours after LPS application, compared with the number at baseline (p < 0.05). Gene expressions of tumor necrosis factor-α (Tnf-α) and forkhead box protein p3 (Foxp3) was also significantly higher in the SPF mice than in the GF mice at 3 hours after LPS application (p < 0.05). The number of neutrophils peaked at 24 hours in both GF and SPF mice. Conclusions: LPS-exposed SPF mice exhibited increases in the number of CD4+ T cells and in Tnf-α and Foxp3 gene expression in periodontal tissue compared with LPS-exposed GF mice.",
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AU - Fukuhara, Daiki

AU - Irie, Koichiro

AU - Uchida, Yoko

AU - Kataoka, Kota

AU - Akiyama, Kentaro

AU - Ekuni, Daisuke

AU - Tomofuji, Takaaki

AU - Morita, Manabu

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N2 - Background: Commensal flora are involved in the appropriate development of the mature immune system. However, it is unclear how commensal flora contribute to immune responses against periodontal pathogens, including the response to lipopolysaccharide (LPS). The purpose of this study was to evaluate the expression of immune responses after topical application of LPS in germ-free (GF) and specific-pathogen-free (SPF) mice. Methods: GF and SPF mice at 8 weeks of age were randomly divided into four groups each: a baseline group (n = 4/group) and three experimental groups (n = 6/group). Experimental groups received topical application of Porphyromonas gingivalis LPS (10 μg/μL) into the palatal gingival sulcus. Sampling was performed before LPS application (baseline) and at 3, 24, or 72 hours after LPS application. The numbers of neutrophils, CD4+, and CD8+ T cells in periodontal tissue were evaluated by immunohistochemistry. Expression of genes encoding cytokines, chemokines, and a transcription factor was determined by real-time PCR. Results: SPF mice, but not GF mice, showed an increased number of CD4+ T cells in the periodontal tissue at 3 hours after LPS application, compared with the number at baseline (p < 0.05). Gene expressions of tumor necrosis factor-α (Tnf-α) and forkhead box protein p3 (Foxp3) was also significantly higher in the SPF mice than in the GF mice at 3 hours after LPS application (p < 0.05). The number of neutrophils peaked at 24 hours in both GF and SPF mice. Conclusions: LPS-exposed SPF mice exhibited increases in the number of CD4+ T cells and in Tnf-α and Foxp3 gene expression in periodontal tissue compared with LPS-exposed GF mice.

AB - Background: Commensal flora are involved in the appropriate development of the mature immune system. However, it is unclear how commensal flora contribute to immune responses against periodontal pathogens, including the response to lipopolysaccharide (LPS). The purpose of this study was to evaluate the expression of immune responses after topical application of LPS in germ-free (GF) and specific-pathogen-free (SPF) mice. Methods: GF and SPF mice at 8 weeks of age were randomly divided into four groups each: a baseline group (n = 4/group) and three experimental groups (n = 6/group). Experimental groups received topical application of Porphyromonas gingivalis LPS (10 μg/μL) into the palatal gingival sulcus. Sampling was performed before LPS application (baseline) and at 3, 24, or 72 hours after LPS application. The numbers of neutrophils, CD4+, and CD8+ T cells in periodontal tissue were evaluated by immunohistochemistry. Expression of genes encoding cytokines, chemokines, and a transcription factor was determined by real-time PCR. Results: SPF mice, but not GF mice, showed an increased number of CD4+ T cells in the periodontal tissue at 3 hours after LPS application, compared with the number at baseline (p < 0.05). Gene expressions of tumor necrosis factor-α (Tnf-α) and forkhead box protein p3 (Foxp3) was also significantly higher in the SPF mice than in the GF mice at 3 hours after LPS application (p < 0.05). The number of neutrophils peaked at 24 hours in both GF and SPF mice. Conclusions: LPS-exposed SPF mice exhibited increases in the number of CD4+ T cells and in Tnf-α and Foxp3 gene expression in periodontal tissue compared with LPS-exposed GF mice.

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KW - Lipopolysaccharide

KW - Microbiota

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