Recombinant fimbriae protein of Porphyromonas gingivalis induces an inflammatory response via the TLR4/NF‑κB signaling pathway in human peripheral blood mononuclear cells

Cai,Jing; Chen,Jiangman; Guo,Huanxu; Pan,Yaping; Zhang,Yibo; Zhao,Wei; Li,Xin; Li,Yonggang

doi:10.3892/ijmm.2019.4069

Recombinant fimbriae protein of Porphyromonas gingivalis induces an inflammatory response via the TLR4/NF‑κB signaling pathway in human peripheral blood mononuclear cells

Authors:
- Jing Cai
- Jiangman Chen
- Huanxu Guo
- Yaping Pan
- Yibo Zhang
- Wei Zhao
- Xin Li
- Yonggang Li
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Affiliations: Department of Periodontics and Oral Biology, School of Stomatology, Jinzhou Medical University, Jinzhou, Liaoning 121000, P.R. China, Department of Periodontics and Oral Biology, School of Stomatology, China Medical University, Shenyang, Liaoning 110002, P.R. China, Department of Pathogeny Biology, Jinzhou Medical University, Jinzhou, Liaoning 121000, P.R. China
Published online on: January 21, 2019 https://doi.org/10.3892/ijmm.2019.4069
Pages: 1430-1440
Copyright: © Cai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Porphyromonas gingivalis (P. gingivalis) is a periodontal pathogen that may accumulate with other organisms in subgingival plaque biofilms and is associated with periodontal disease. P. gingivalis fimbriae (FimA) is a filamentous structure on the surface of bacteria that is closely associated with bacterial adhesion to and colonization of host tissues, and serves an essential role in biofilm formation. The present study aimed to construct P. gingivalis FimA prokaryotic expression plasmids, purify a FimA fusion protein and explore the effect of a recombinant FimA protein on the inflammatory response in human peripheral blood mononuclear cells (PBMCs). P. gingivalis FimA prokaryotic expression plasmids were constructed by gene cloning and recombination technology. SDS‑PAGE was used to evaluate the purified recombinant FimA protein. The cell proliferation rate and inflammatory cytokine expression of PBMCs treated with the FimA fusion protein with or without transfection with toll‑like receptor 4 (TLR4) small interfering (si)RNA were detected by CCK‑8 assays and ELISAs, respectively. The expression levels of TLR4, nuclear factor kappa‑light‑chain‑enhancer of activated B cells (NF‑κB) and myeloid differentiation primary response 88 (MyD88) in PBMCs were detected by western blot analysis and reverse transcription quantitative polymerase chain reaction. A FimA fusion protein with high purity was obtained. FimA fusion protein treatment significantly increased PBMC proliferation and promoted the release of tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑6, matrix metalloproteinase (MMP)‑8 and MMP‑9 in PBMCs. TLR4 interference reversed the effects of the FimA fusion protein on PBMC proliferation and inflammatory cytokine release. The expression levels of TLR4, NF‑κB and MyD88 in PBMCs were significantly increased following treatment with the FimA fusion protein, while the expression levels of these genes at the mRNA and protein levels decreased significantly in PBMCs following FimA fusion protein treatment and TLR4 interference. The FimA fusion protein increased PBMC proliferation and promoted the release of the inflammatory cytokines TNF‑α, IL‑6, MMP‑8 and MMP‑9 via the TLR4/NF‑κB signaling pathway. FimA may serve as a promising therapeutic strategy for periodontal disease.

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1	Pennisi E: A mouthful of microbes. Science. 307:1899–1901. 2005. View Article : Google Scholar : PubMed/NCBI
2	Schutzhold S, Kocher T, Biffar R, Hoffmann T, Schmidt CO, Micheelis W, Jordan R and Holtfreter B: Changes in prevalence of periodontitis in two German population-based studies. J Clin Periodontol. 42:121–130. 2015. View Article : Google Scholar
3	Hasiuk P, Hasiuk N, Kindiy D, Ivanchyshyn V, Kalashnikov D and Zubchenko S: Characteristics of cellular composition of periodontal pockets. Interv Med Appl Sci. 8:172–177. 2016.
4	Aimetti M, Perotto S, Castiglione A, Mariani GM, Ferrarotti F and Romano F: Prevalence of periodontitis in an adult population from an urban area in North Italy: Findings from a cross-sectional population-based epidemiological survey. J Clin Periodontol. 42:622–631. 2015. View Article : Google Scholar : PubMed/NCBI
5	Souza JA, Medeiros MC, Rocha FR, de Aquino SG, Ávila-Campos MJ, Spolidorio LC, Zamboni DS and Graves DT: Role of NOD2 and RIP2 in host-microbe interactions with Gram-negative bacteria: Insights from the periodontal disease model. Innate Immun. 22:598–611. 2016. View Article : Google Scholar : PubMed/NCBI
6	Haffajee AD and Socransky SS: Microbial etiological agents of destructive periodontal diseases. Periodontol. 5:78–111. 2000. View Article : Google Scholar
7	Tsuda K, Furuta N, Inaba H, Kawai S, Hanada K, Yoshimori T and Amano A: Functional analysis of alpha5beta1 integrin and lipid rafts in invasion of epithelial cells by Porphyromonas gingivalis using fluorescent beads coated with bacterial membrane vesicles. Cell Struct Funct. 33:123–132. 2008. View Article : Google Scholar : PubMed/NCBI
8	Bozkurt SB, Hakki SS, Hakki EE, Durak Y and Kantarci A: Porphyromonas gingivalis lipopolysaccharide induces a pro-inflammatory human gingival fibroblast phenotype. Inflammation. 40:144–153. 2017. View Article : Google Scholar
9	Zheng C, Wu J and Xie H: Differential expression and adherence of Porphyromonas gingivalis FimA genotypes. Mol Oral Microbiol. 26:388–395. 2011. View Article : Google Scholar : PubMed/NCBI
10	Dashper SG, Mitchell HL, Seers CA, Gladman SL, Seemann T, Bulach DM, Chandry PS, Cross KJ, Cleal SM and Reynolds EC: Porphyromonas gingivalis uses specific domain rearrangements and allelic exchange to generate diversity in surface virulence factors. Front Microbiol. 8:482017. View Article : Google Scholar :
11	Seo NR, Ji MK, Park SW, Lee K, Bae JC, Yun KD, Park C, Oh GJ, Kim JH and Lim HP: Effect on adhesion of porphyromonas gingivalis by titanium nitride sputter coating or plasma nitriding of titanium. J Nanosci Nanotechnol. 17:2633–2636. 2017. View Article : Google Scholar
12	Feng X, Zhang L, Xu L, Meng H, Lu R, Chen Z, Shi D and Wang X: Detection of eight periodontal microorganisms and distribution of Porphyromonas gingivalis fimA genotypes in Chinese patients with aggressive periodontitis. J Periodontol. 85:150–159. 2014. View Article : Google Scholar
13	Zhou W, Zhang X, Zhu CL, He ZY, Liang JP and Song ZC: Melatonin receptor agonists as the 'perioceutics' agents for periodontal disease through modulation of porphyromonas gingivalis virulence and inflammatory response. PLoS One. 11:e1664422016. View Article : Google Scholar
14	Ishikado A, Uesaki S, Suido H, Nomura Y, Sumikawa K, Maeda M, Miyauchi M, Takata T and Makino T: Human trial of liposomal lactoferrin supplementation for periodontal disease. Biol Pharm Bull. 33:1758–1762. 2010. View Article : Google Scholar : PubMed/NCBI
15	Basic A, Alizadehgharib S, Dahlén G and Dahlgren U: Hydrogen sulfide exposure induces NLRP3 inflammasome-dependent IL-1β and IL-18 secretion in human mononuclear leukocytes in vitro. Clin Exp Dent Res. 3:115–120. 2017. View Article : Google Scholar
16	Cutando A, Montero J, Gómez-de Diego R, Ferrera MJ and Lopez-Valverde A: Effect of topical application of melatonin on serum levels of C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in patients with type 1 or type 2 diabetes and periodontal disease. J Clin Exp Dent. 7:e628–e633. 2015.PubMed/NCBI
17	Otenio CC, Fonseca I, Martins MF, Ribeiro LC, Assis NM, Ferreira AP and Ribeiro RA: Expression of IL-1β, IL-6, TNF-α, and iNOS in pregnant women with periodontal disease. Genet Mol Res. 11:4468–4478. 2012. View Article : Google Scholar : PubMed/NCBI
18	Marcaccini AM, Novaes AB Jr, Meschiari CA, Souza SL, Palioto DB, Sorgi CA, Faccioli LH, Tanus-Santos JE and Gerlach RF: Circulating matrix metalloproteinase-8 (MMP-8) and MMP-9 are increased in chronic periodontal disease and decrease after non-surgical periodontal therapy. Clin Chim Acta. 409:117–122. 2009. View Article : Google Scholar : PubMed/NCBI
19	Zhou Q and Amar S: Identification of signaling pathways in macrophage exposed to Porphyromonas gingivalis or to its purified cell wall components. J Immunol. 179:7777–7790. 2007. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
21	Wang D, Tao K, Xion J, Xu S, Jiang Y, Chen Q and He S: TAK-242 attenuates acute cigarette smoke-induced pulmonary inflammation in mouse via the TLR4/NF-κB signaling pathway. Biochem Biophys Res Commun. 472:508–515. 2016. View Article : Google Scholar : PubMed/NCBI
22	Sanz M, van Winkelhoff AJ, Herrera D, Dellemijn-Kippuw N, Simón R and Winkel E: Differences in the composition of the subgingival microbiota of two periodontitis populations of different geographical origin. A comparison between Spain and The Netherlands. Eur J Oral Sci. 108:383–392. 2000. View Article : Google Scholar : PubMed/NCBI
23	Sharma A, Honma K, Evans RT, Hruby DE and Genco RJ: Oral immunization with recombinant Streptococcus gordonii expressing porphyromonas gingivalis FimA domains. Infect Immun. 69:2928–2934. 2001. View Article : Google Scholar : PubMed/NCBI
24	Mogensen TH: Pathogen recognition and inflammatory signaling in innate immune defenses. Clin Microbiol Rev. 22:240–273. 2009. View Article : Google Scholar : PubMed/NCBI
25	Zhou Q, Desta T, Fenton M, Graves DT and Amar S: Cytokine profiling of macrophages exposed to Porphyromonas gingivalis, its lipopolysaccharide, or its FimA protein. Infect Immun. 73:935–943. 2005. View Article : Google Scholar : PubMed/NCBI
26	Suwatanapongched P, Surarit R, Srisatjaluk R and Offenbacher S: Translocation of Porphyromonas gingivalis infected mono-cytes and associated cellular responses. Asian Pac J Allergy Immunol. 28:192–199. 2010.PubMed/NCBI
27	Nozoe K, Sanui T, Takeshita M, Fukuda T, Haraguchi A, Aida Y and Nishimura F: Innate immune-stimulatory activity of Porphyromonas gingivalis fimbriae is eliminated by phase separation using Triton X-114. J Immunol Methods. 441:31–38. 2017. View Article : Google Scholar
28	Yun PL, Decarlo AA, Collyer C and Hunter N: Hydrolysis of interleukin-12 by Porphyromonas gingivalis major cysteine proteinases may affect local gamma interferon accumulation and the Th1 or Th2 T-cell phenotype in periodontitis. Infect Immun. 69:5650–5660. 2001. View Article : Google Scholar : PubMed/NCBI
29	Chaudhuri S, Pratap S, Paromov V, Li Z, Mantri CK and Xie H: Identification of a diguanylate cyclase and its role in Porphyromonas gingivalis virulence. Infect Immun. 82:2728–2735. 2014. View Article : Google Scholar : PubMed/NCBI
30	Mantri CK, Chen CH, Dong X, Goodwin JS, Pratap S, Paromov V and Xie H: Fimbriae-mediated outer membrane vesicle production and invasion of Porphyromonas gingivalis. Microbiologyopen. 4:53–65. 2015. View Article : Google Scholar :
31	Ogawa T, Asai Y, Hashimoto M and Uchida H: Bacterial fimbriae activate human peripheral blood monocytes utilizing TLR2, CD14 and CD11a/CD18 as cellular receptors. Eur J Immunol. 32:2543–2550. 2002. View Article : Google Scholar : PubMed/NCBI
32	Deng B, Fang F, Yang T, Yu Z, Zhang B and Xie X: Ghrelin inhibits AngII-induced expression of TNF-α, IL-8, MCP-1 in human umbilical vein endothelial cells. Int J Clin Exp Med. 8:579–588. 2015.
33	Rodrigues PH, Reyes L, Chadda AS, Bélanger M, Wallet SM, Akin D, Dunn W Jr and Progulske-Fox A: Porphyromonas gingivalis strain specific interactions with human coronary artery endothelial cells: A comparative study. PLoS One. 7:e526062012. View Article : Google Scholar
34	Nagano K, Hasegawa Y, Abiko Y, Yoshida Y, Murakami Y and Yoshimura F: Porphyromonas gingivalis FimA Fimbriae: Fimbrial assembly by fimA alone in the fim gene cluster and differential antigenicity among fimA genotypes. PLoS One. 7:e437222012. View Article : Google Scholar : PubMed/NCBI
35	Cochran DL: Inflammation and bone loss in periodontal disease. J Periodontol. 79:1569–1576. 2008. View Article : Google Scholar : PubMed/NCBI
36	Ikezawa I, Tai H, Shimada Y, Komatsu Y, Galicia JC and Yoshie H: Imbalance between soluble tumour necrosis factor receptors type 1 and 2 in chronic periodontitis. J Clin Periodontol. 32:1047–1054. 2005. View Article : Google Scholar : PubMed/NCBI
37	Chen Y, Zhou R, Yi Z, Li Y, Fu Y, Zhang Y, Li P, Li X and Pan Y: Porphyromonas gingivalis induced inflammatory responses and promoted apoptosis in lung epithelial cells infected with H1N1 via the Bcl-2/Bax/Caspase-3 signaling pathway. Mol Med Rep. 18:97–104. 2018.PubMed/NCBI
38	Graves DT, Naguib G, Lu H, Desta T and Amar S: Porphyromonas gingivalis fimbriae are pro-inflammatory but do not play a prominent role in the innate immune response to P. gingivalis. J Endotoxin Res. 11:13–18. 2005. View Article : Google Scholar : PubMed/NCBI
39	Hajishengallis G, Sojar HT, Sharma A, Martin M, Denardini E and Genco RJ: TLR4-dependent upregulation of costimulatory molecules in macrophages by p. gingivalis FimA. J Dent Res. 81:A532002.
40	Hajishengallis G, Martin M, Schifferle RE and Genco RJ: Counteracting interactions between lipopolysaccharide molecules with differential activation of toll-like receptors. Infect Immun. 70:6658–6664. 2002. View Article : Google Scholar : PubMed/NCBI