Secreted gingipains from Porphyromonas gingivalis increase permeability in human cerebral microvascular endothelial cells through intracellular degradation of tight junction proteins
Open accessAbstract
Despite a clear correlation between the infiltration of periodontal pathogens in the brain and cognitive decline in Alzheimer's disease (AD), the precise mechanism underlying bacteria crossing the blood-brain barrier (BBB) remains unclear. The periodontal pathogen Porphyromonas gingivalis produces a unique class of cysteine proteases termed gingipains. Gingipains appear to be key virulence factors that exacerbate sporadic AD. We herein report that gingipains are involved in increasing permeability of hCMEC/D3 cell monolayer, human cerebral microvascular endothelial cell lines, through degradation of tight junction proteins including Zonula occludens-1 (ZO-1) and occludin. There was a significant decrease in the mean protein levels of ZO-1 and occludin after infection of hCMEC/D3 cells with wild-type (WT) P. gingivalis. However, infection of these cells with a gingipain-deficient P. gingivalis strain showed significantly lower reduction of the mean protein levels of either ZO-1 and occludin, compared to the WT strain. Similar results were obtained after treatment with culture supernatant from WT and gingipain-deficient P. gingivalis strains. In vitro digestion of human recombinant ZO-1 and occludin by WT P. gingivalis culture supernatant in the absence or presence of gingipain inhibitors indicated that gingipains directly degraded these tight junction proteins. A close immunohistochemical examination using anti-gingipain antibody further revealed that gingipains localized in the cytosol and nuclei of hCMEC/D3 cells after infection with WT P. gingivalis and treatment with its culture supernatant. Furthermore, intracellular localization of outer membrane vesicles (OMVs) bound gingipains from WT P. gingivalis and OMV-induced degradation of ZO-1 and occludin were also observed in hCMEC/D3 cells. Thus, the delivery of gingipains into the cerebral microvascular endothelial cells, probably through OMV, may be responsible for the BBB damage through intracellular degradation of ZO-1 and occludin.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Gingipains are potent proteolytic enzymes produced by Pg that degrade tight junction proteins of human endothelial cells.
“Gingipains are potent proteolytic enzymes produced by Pg that degrade tight junction proteins of human endothelial cells [3,4].”
From Oral Microbially-Induced Small Extracellular Vesicles Cross the Blood–Brain Barrier (2024) · cites this paper as [3]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Dominy, S. S., Lynch, C., Ermini, F., Benedyk, M., Marczyk, A., Konradi, A., Nguyen, M., Haditsch, U., Raha, D., Griffin, C., et al. (2019) · Science Advances · doi:10.1126/sciadv.aau3333 · Cited by 1,966 · see citing works
Ilievski, V., Zuchowska, P. K., Green, S. J., Toth, P. T., Ragozzino, M. E., Le, K., Aljewari, H. W., O’Brien-Simpson, N. M., Reynolds, E. C., & Watanabe, K. (2018) · PLOS ONE · doi:10.1371/journal.pone.0204941 · Cited by 372 · see citing works
Flagged first, then by citations
Research works printed but not linked: 33
- Ben Lagha, A., Pellerin, G., Vaillancourt, K., Grenier, D., 2021. Effects of a tart cherry (Prunus cerasus L.) phenolic extract on Porphyromonas gingivalis and its ability to impair the oral epithelial barrier. PLoS One 16, e0246194. https://doi.org/10.1371/ journal.pone.0246194. · DOI
- Ding, Y., Ren, J., Yu, H., Yu, W., Zhou, Y., 2018. Porphyromonas gingivalis, a periodontitis causing bacterium, induces memory impairment and age-dependent neuroinflammation in mice. Immun. Ageing 15, 6. https://doi.org/10.1186/s12979- 017-0110-7. · DOI
- Falconer, A.M.D., Chan, C.M., Gray, J., Nagashima, I., Holland, R.A., Shimizu, H., Pickford, A.R., Rowan, A.D., Wilkinson, D.J., 2019. Collagenolytic matrix metalloproteinases antagonize proteinase-activated receptor-2 activation, providing insights into extracellular matrix turnover. J. Biol. Chem. 294, 10266-10277. https://doi.org/10.1074/jbc.RA119.006974. · DOI
- Furuta, N., Tsuda, K., Omori, H., Yoshimori, T., Yoshimura, F., Amano, A., 2009. Porphyromonas gingivalis outer membrane vesicles enter human epithelial cells via an endocytic pathway and are sorted to lysosomal compartments. Infect. Immun. 77, 4187-4196. https://doi.org/10.1128/IAI.00009-09. · DOI
- Furutama, D., Matsuda, S., Yamawaki, Y., Hatano, S., Okanobu, A., Memida, T., Oue, H., Fujita, T., Ouhara, K., Kajiya, M., Mizuno, N., Kanematsu, T., Tsuga, K., Kurihara, H., 2020. IL-6 induced by periodontal inflammation causes neuroinflammation and disrupts the blood-brain barrier. Brain Sci. 10, 679. https://doi.org/10.3390/ brainsci10100679. · DOI
- Guo, Y., Nguyen, K.A., Potempa, J., 2010. Dichotomy of gingipains action as virulence factors: from cleaving substrates with the precision of a surgeon's knife to a meat chopper-like brutal degradation of proteins. Periodontol 54, 15-44. https://doi.org/ 10.1111/j.1600-0757.2010.00377.x. · DOI
- Ha, J.Y., Choi, S.Y., Lee, J.H., Hong, S.H., Lee, H.J., 2020. Delivery of periodontopathogenic extracellular vesicles to brain monocytes and microglial IL-6 promotion by RNA Cargo. Front. Mol. Biosci. 7, 596366. https://doi.org/10.3389/ fmolb.2020.596366. · DOI
- Haditsch, U., Roth, T., Rodriguez, L., Hancock, S., Cecere, T., Nguyen, M., Arastu- Kapur, S., Broce, S., Raha, D., Lynch, C.C., Holsinger, L.J., Dominy, S.S., Ermini, F., 2020. Alzheimer's disease-like neurodegeneration in Porphyromonas gingivalis infected neurons with persistent expression of active gingipains. J. Alzheimers Dis. 75, 1361-1376. https://doi.org/10.3233/JAD-200393. · DOI
- Han, E.C., Choi, S.Y., Lee, Y., Park, J.W., Hong, S.H., Lee, H.J., 2019. Extracellular RNAs in periodontopathogenic outer membrane vesicles promote TNF-α production in human macrophages and cross the blood-brain barrier in mice. Faseb. J. 33, 13412-13422. https://doi.org/10.1096/fj.201901575R. · DOI
- Holmer, J., Eriksdotter, M., Schultzberg, M., Pussinen, P.J., Buhlin, K., 2018. Association between periodontitis and risk of Alzheimer's disease, mild cognitive impairment and subjective cognitive decline: a case-control study. J. Clin. Periodontol. 45, 1287-1298. https://doi.org/10.1111/jcpe.13016. · DOI
- Huang, W., Zeng, F., Gu, Y., Jiang, M., Zhang, X., Yan, X., Kadowaki, T., Mizutani, S., Kashiwazaki, H., Ni, J., Wu, Z., 2021. Porphyromonas gingivalis infection induces synaptic failure via increased IL-1β production in leptomeningeal cells. J. Alzheimers Dis. 83, 665-681. https://doi.org/10.3233/JAD-210031. · DOI
- Ide, M., Harris, M., Stevens, A., Sussams, R., Hopkins, V., Culliford, D., Fuller, J., Ibbett, P., Raybould, R., Thomas, R., Puenter, U., Teeling, J., Perry, V.H., Holmes, C., 2016. Periodontitis and cognitive decline in Alzheimer's disease. PLoS One 11, e0151081. https://doi.org/10.1371/journal.pone.0151081. · DOI · Paper page
- Inaba, H., Sugita, H., Kuboniwa, M., Iwai, S., Hamada, M., Noda, T., Morisaki, I., Lamont, R.J., Amano, A., 2014. Porphyromonas gingivalis promotes invasion of oral squamous cell carcinoma through induction of proMMP9 and its activation. Cell Microbiol. 16, 131-145. https://doi.org/10.1111/cmi.12211. · DOI
- Kadowaki, T., Baba, A., Abe, N., Takii, R., Hashimoto, M., Tsukuba, T., Okazaki, S., Suda, Y., Asao, T., Yamamoto, K., 2004. Suppression of pathogenicity of Porphyromonas gingivalis by newly developed gingipain inhibitors. Mol. Pharmacol. 66, 1599-1606. https://doi.org/10.1124/mol.104.004366. · DOI
- Kamer, A.R., Craig, R.G., Dasanayake, A.P., Brys, M., Glodzik-Sobanska, L., de Leon, M. J., 2008a. Inflammation and Alzheimer's disease: possible role of periodontal diseases. Alzheimers Dement 4, 242-250. https://doi.org/10.1016/j. jalz.2007.08.004. · DOI
- Kamer, A.R., Dasanayake, A.P., Craig, R.G., Glodzik-Sobanska, L., Bry, M., de Leon, M.J., 2008b. Alzheimer's disease and peripheral infections: the possible contribution from periodontal infections, model and hypothesis. J. Alzheimers Dis. 13, 437-449. https://doi.org/10.3233/JAD-2008-13408. · DOI
- Li, X.F., Zhang, X.J., Zhang, C., Wang, L.N., Li, Y.R., Zhang, Y., He, T.T., Zhu, X.Y., Cui, L. L., Gao, B.L., 2018. Ulinastatin protects brain against cerebral ischemia/reperfusion injury through inhibiting MMP-9 and alleviating loss of ZO-1 and occludin proteins in mice. Exp. Neurol. 302, 68-74. https://doi.org/10.1016/j. expneurol.2017.12.016. · DOI
- Liu, Y., Wu, Z., Nakanishi, Y., Ni, J., Hayashi, Y., Takayama, F., Zhou, Y., Kadowaki, T., Nakanishi, H., 2017. Infection of microglia with Porphyromonas gingivalis promotes cell migration and an inflammatory response through the gingipain-mediated activation of protease-activated receptor-2 in mice. Sci. Rep. 7, 11759. https://doi. org/10.1038/s41598-017-12173-1. · DOI
- Lu, D.Y., Yu, W.H., Yeh, W.L., Tang, C.H., Leung, Y.M., Wong, K.L., Chen, Y.F., Lai, C.H., Fu, W.M., 2009. Hypoxia-induced matrix metalloproteinase-13 expression in astrocytes enhances permeability of brain endothelial cells. J. Cell. Physiol. 220, 163-173. https://doi.org/10.1002/jcp.21746. · DOI
- Nakanishi, H., Nonaka, S., Wu, Z., 2020. Microglial Cathepsin B and Porphyromonas gingivalis gingipains as potential therapeutic targets for sporadic Alzheimer's disease. CNS Neurol. Disord. -Drug Targets 19, 495-502. https://doi.org/10.2174/ 1871527319666200708125130. · DOI
- Nakao, R., Takashiba, S., Kosono, S., Yoshida, M., Watanabe, H., Ohnishi, M., Senpuku, H., 2014. Effect of Porphyromonas gingivalis outer membrane vesicles on gingipain-mediated detachment of cultured oral epithelial cells and immune responses. Microbes Infect. 16, 6-16. https://doi.org/10.1016/j. micinf.2013.10.005. · DOI
- Nara, P.L., Sindelar, D., Penn, M.S., Potempa, J., Griffin, W.S.T., 2021. Porphyromonas gingivalis outer membrane vesicles as the major driver of and explanation for neuropathogenesis, the cholinergic hypothesis, iron dyshomeostasis, and salivary lactoferrin in Alzheimer's disease. J. Alzheimers Dis. 82, 1417-1450. https://doi. org/10.3233/JAD-210448. · DOI · Paper page
- Nonaka, S., Nakanishi, H., 2020. Secreted gingipains from Porphyromonas gingivalis induce microglia migration through endosomal signaling by protease-activated receptor 2. Neurochem. Int. 140, 104840. https://doi.org/10.1016/j. neuint.2020.104840. · DOI
- Okamura, H., Hirota, K., Yoshida, K., Weng, Y., He, Y., Shiotsu, N., Ikegame, M., Uchida- Fukuhara, Y., Tanai, A., Guo, J., 2021. Outer membrane vesicles of Porphyromonas gingivalis: novel communication tool and strategy. Jpn. Dent. Sci. Rev. 57, 138-146. https://doi.org/10.1016/j.jdsr.2021.07.003. · DOI
- Poole, S., Singhrao, S.K., Kesavalu, L., Curtis, M.A., Crean, S., 2013. Determining the presence of periodontopathic virulence factors in short-term postmortem Alzheimer's disease brain tissue. J. Alzheimers Dis. 36, 665-677. https://doi.org/ 10.3233/JAD-121918. · DOI
- Sato, N., Matsumoto, T., Kawaguchi, S., Seya, K., Matsumiya, T., Ding, J., Aizawa, T., Imaizumi, T., 2021. Porphyromonas gingivalis lipopolysaccharide induces interleukin- 6 and c-c motif chemokine ligand 2 expression in cultured hCMEC/D3 human brain microvascular endothelial cells. Gerodontology. https://doi.org/10.1111/ ger.12545. · DOI
- Seyama, M., Yoshida, K., Yoshida, K., Fujiwara, N., Ono, K., Eguchi, T., Kawai, H., Guo, J., Weng, Y., Haoze, Y., Uchibe, K., Ikegame, M., Sasaki, A., Nagatsuka, H., Okamoto, K., Okamura, H., Ozaki, K., 2020. Outer membrane vesicles of Porphyromonas gingivalis attenuate insulin sensitivity by delivering gingipains to the liver. Biochim. Biophys. Acta (BBA) -Mol. Basis Dis. 1866, 165731. https://doi.org/ 10.1016/j.bbadis.2020.165731. · DOI
- Singhrao, S.K., Harding, A., Simmons, T., Robinson, S., Kesavalu, L., Crean, S., 2014. Oral inflammation, tooth loss, risk factors, and association with progression of Alzheimer's disease. J. Alzheimers Dis. 42, 723-737. https://doi.org/10.3233/JAD- 140387. · DOI
- Singhrao, S.K., Olsen, I., 2018. Are Porphyromonas gingivalis outer membrane vesicles microbullets for sporadic Alzheimer's disease manifestation? J. Alzheimers Dis. Rep. 2, 219-228. https://doi.org/10.3233/ADR-180080. · DOI
- Tsuzuno, T., Takahashi, N., Yamada-Hara, M., Yokoji-Takeuchi, M., Sulijaya, B., Aoki- Nonaka, Y., Matsugishi, A., Katakura, K., Tabeta, K., Yamazaki, K., 2021. Ingestion of Porphyromonas gingivalis exacerbates colitis via intestinal epithelial barrier disruption in mice. J. Periodontal. Res. 56, 275-288. https://doi.org/10.1111/ jre.12816. · DOI
- Wu, Z., Nakanishi, H., 2014. Connection between periodontitis and Alzheimer's disease: possible roles of microglia and leptomeningeal cells. J. Pharmacol. Sci. 126, 8-13. https://doi.org/10.1254/jphs.14r11cp. · DOI
- Zhang, Z., Liu, D., Liu, S., Zhang, S., Pan, Y., 2020. The role of Porphyromonas gingivalis outer membrane vesicles in periodontal disease and related systemic diseases. Front. Cell. Infect. Microbiol. 10, 585917. https://doi.org/10.3389/fcimb.2020.585917. · DOI
- Zhou, Q., Wang, Y.W., Ni, P.F., Chen, Y.N., Dong, H.Q., Qian, Y.N., 2018. Effect of tryptase on mouse brain microvascular endothelial cells via protease-activated receptor 2. J. Neuroinflammation 15, 248. https://doi.org/10.1186/s12974-018- 1287-1. · DOI
Cited by
Sorted by citations
Oral Microbially-Induced Small Extracellular Vesicles Cross the Blood–Brain Barrier
Elashiry, M., Carroll, A., Yuan, J., Liu, Y., Hamrick, M., Cutler, C. W., Wang, Q., & Elsayed, R. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25084509 · Cited by 21 · see citing works
“A role for periodontitis (PD) in the pathogenesis of AD has recently been proposed by several studies [3–13].”
Result
- Authors
- Nonaka, S., Kadowaki, T., & Nakanishi, H.
- Affiliations
- Yasuda Women's University (JP)
- Nagasaki University (JP)
- Corresponding author
- Nonaka, Saori
- Nakanishi, Hiroshi
- Venue
- Neurochemistry International
- Publisher
- Elsevier BV
- Published
- 2022-03
- Volume
- 154
- Pages
- 105282
- ISSN
- 0197-0186 (ISSN-L)
- OpenAlex
- W4205248667
- Cited by
- 99
- References
- 2
- Citations by year
- 2026: 24 · 2025: 32 · 2024: 21 · 2023: 16 · 2022: 6
- FWCI
- 7.20
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Neurochemistry International (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Promotion and Mutual Aid Corporation for Private Schools of Japan
- Japan Society for the Promotion of Science (22K15287, JP21K06383)
- Topics
- Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
- Health Sciences › Dentistry › Periodontics › Oral microbiology and periodontitis research
- Life Sciences › Immunology and Microbiology › Microbiology › Bacterial Infections and Vaccines
- Keywords
- Porphyromonas gingivalis, gingipains, blood-brain barrier permeability, tight junction proteins, ZO-1, occludin, brain microvascular endothelial cells, Alzheimer's disease, periodontal pathogens, outer membrane vesicles, cysteine protease, bacterial virulence factors
- MeSH terms
- Gingipain Cysteine Endopeptidases, Humans, Permeability, Porphyromonas gingivalis, Adhesins, Bacterial, Endothelial Cells, Tight Junction Proteins
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
