Effect of toxins from different periodontitis-associated bacteria on human platelet function.
CD62P
Gram‐negative bacteria
lipopolysaccharides
periodontitis
platelets
vasodilator‐stimulated phosphoprotein
Journal
Molecular oral microbiology
ISSN: 2041-1014
Titre abrégé: Mol Oral Microbiol
Pays: Denmark
ID NLM: 101524770
Informations de publication
Date de publication:
26 Jul 2024
26 Jul 2024
Historique:
revised:
22
06
2024
received:
18
10
2023
accepted:
14
07
2024
medline:
26
7
2024
pubmed:
26
7
2024
entrez:
26
7
2024
Statut:
aheadofprint
Résumé
Periodontitis is caused by a dysbiosis of oral bacteria resulting in alveolar bone destruction and teeth loss. The role of platelets in pathogenesis of periodontitis is a subject of research. The release of toxins from periodontitis-associated bacteria may influence platelet function and contribute to the modulation of hemostatic or inflammatory responses. Therefore, we explored platelet function upon exposure to defined toxins: leukotoxin A from Aggregatibacter actinomycetemcomitans (LtxA), a synthetic version of the C14-Tri-LAN-Gly peptide from Fusobacterium nucleatum (C14), and lipopolysaccharides from Porphyromonas gingivalis (LPS). Light transmission aggregometry was performed after the addition of toxins to platelet-rich plasma in different doses. Flow cytometry was used to identify inhibitory effects of toxins by measuring phosphorylation of the vaso-dilator-stimulated phosphoprotein or to identify activating effects by the detection of CD62P expression. The release of chemokines derived from washed platelets was determined by immunoassays. Collagen-induced threshold aggregation values were diminished upon incubation with LtxA and C14, accompanied with an increase of vaso-dilator-stimulated phosphoprotein (VASP) phosphorylation, indicating platelet inhibition. In contrast, LPS did not affect aggregation but slightly enhanced CD62P expression under co-stimulation with low-dose thrombin pointing to slight platelet activation. The three toxins did not relevantly influence the secretion of chemokines. Although weak, the investigated toxins differently influenced human platelet function. LtxA and C14 mediated inhibitory effects, whereas LPS contributed to a slight activation of platelets. Further analysis of specific cellular responses mediated by bacterial toxins may render novel targets and suggestions for the treatment of periodontitis.
Sections du résumé
BACKGROUND
BACKGROUND
Periodontitis is caused by a dysbiosis of oral bacteria resulting in alveolar bone destruction and teeth loss. The role of platelets in pathogenesis of periodontitis is a subject of research. The release of toxins from periodontitis-associated bacteria may influence platelet function and contribute to the modulation of hemostatic or inflammatory responses. Therefore, we explored platelet function upon exposure to defined toxins: leukotoxin A from Aggregatibacter actinomycetemcomitans (LtxA), a synthetic version of the C14-Tri-LAN-Gly peptide from Fusobacterium nucleatum (C14), and lipopolysaccharides from Porphyromonas gingivalis (LPS).
METHODS
METHODS
Light transmission aggregometry was performed after the addition of toxins to platelet-rich plasma in different doses. Flow cytometry was used to identify inhibitory effects of toxins by measuring phosphorylation of the vaso-dilator-stimulated phosphoprotein or to identify activating effects by the detection of CD62P expression. The release of chemokines derived from washed platelets was determined by immunoassays.
RESULTS
RESULTS
Collagen-induced threshold aggregation values were diminished upon incubation with LtxA and C14, accompanied with an increase of vaso-dilator-stimulated phosphoprotein (VASP) phosphorylation, indicating platelet inhibition. In contrast, LPS did not affect aggregation but slightly enhanced CD62P expression under co-stimulation with low-dose thrombin pointing to slight platelet activation. The three toxins did not relevantly influence the secretion of chemokines.
CONCLUSIONS
CONCLUSIONS
Although weak, the investigated toxins differently influenced human platelet function. LtxA and C14 mediated inhibitory effects, whereas LPS contributed to a slight activation of platelets. Further analysis of specific cellular responses mediated by bacterial toxins may render novel targets and suggestions for the treatment of periodontitis.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2024 The Author(s). Molecular Oral Microbiology published by John Wiley & Sons Ltd.
Références
Assinger, A., Buchberger, E., Laky, M., Esfandeyari, A., Brostjan, C., & Volf, I. (2011). Periodontopathogens induce soluble P‐selectin release by endothelial cells and platelets. Thrombosis Research, 127(1), e20–e26. https://doi.org/10.1016/j.thromres.2010.10.023
Balashova, N. V., Crosby, J. A., Al Ghofaily, L., & Kachlany, S. C. (2006). Leukotoxin confers beta‐hemolytic activity to Actinobacillus actinomycetemcomitans. Infection and Immunity, 74(4), 2015–2021. https://doi.org/10.1128/IAI.74.4.2015‐2021.2006
Bartova, J., Sommerova, P., Lyuya‐Mi, Y., Mysak, J., Prochazkova, J., Duskova, J., Janatova, T., & Podzimek, S. (2014). Periodontitis as a risk factor of atherosclerosis. Journal of Immunology Research, 2014, 636893. https://doi.org/10.1155/2014/636893
Berman, C. L., Yeo, E. L., Wencel‐Drake, J. D., Furie, B. C., Ginsberg, M. H., & Furie, B. (1986). A platelet alpha granule membrane protein that is associated with the plasma membrane after activation. Characterization and subcellular localization of platelet activation‐dependent granule‐external membrane protein. Journal of Clinical Investigation, 78(1), 130–137. https://doi.org/10.1172/JCI112542
Chen, W. A., Fletcher, H. M., Gheorghe, J. D., Oyoyo, U., & Boskovic, D. S. (2020). Platelet plug formation in whole blood is enhanced in the presence of Porphyromonas gingivalis. Molecular Oral Microbiology, 35(6), 251–259. https://doi.org/10.1111/omi.12314
Clemetson, K. J. (2012). Platelets and primary haemostasis. Thrombosis Research, 129(3), 220–224. https://doi.org/10.1016/j.thromres.2011.11.036
Dixon, D. R., & Darveau, R. P. (2005). Lipopolysaccharide heterogeneity: innate host responses to bacterial modification of lipid a structure. Journal of Dental Research, 84(7), 584–595. https://doi.org/10.1177/154405910508400702
Duerschmied, D., Bode, C., & Ahrens, I. (2014). Immune functions of platelets. Thrombosis and Haemostasis, 112(4), 678–691. https://doi.org/10.1160/TH14‐02‐0146
Fujita, A., Oogai, Y., Kawada‐Matsuo, M., Nakata, M., Noguchi, K., & Komatsuzawa, H. (2021). Expression of virulence factors under different environmental conditions in Aggregatibacter actinomycetemcomitans. Microbiology and Immunology, 65(3), 101–114. https://doi.org/10.1111/1348‐0421.12864
Hajishengallis, G., Darveau, R. P., & Curtis, M. A. (2012). The keystone‐pathogen hypothesis. Nature Reviews Microbiology, 10(10), 717–725. https://doi.org/10.1038/nrmicro2873
Herbert, B. A., Novince, C. M., & Kirkwood, K. L. (2016). Aggregatibacter actinomycetemcomitans, a potent immunoregulator of the periodontal host defense system and alveolar bone homeostasis. Molecular Oral Microbiology, 31(3), 207–227. https://doi.org/10.1111/omi.12119
Jockel‐Schneider, Y., Kobsar, A., Stellzig‐Eisenhauer, A., Vogel, U., Störk, S., Frantz, S., Schlagenhauf, U., & Eigenthaler, M. (2018). Wild‐type isolates of Porphyromonas gingivalis derived from periodontitis patients display major variability in platelet activation. Journal of Clinical Periodontology, 45(6), 693–700. https://doi.org/10.1111/jcpe.12895
Joss, A., Adler, R., & Lang, N. P. (1994). Bleeding on probing. A parameter for monitoring periodontal conditions in clinical practice. Journal of Clinical Periodontology, 21(6), 402–408. https://doi.org/10.1111/j.1600‐051x.1994.tb00737.x
Kachlany, S. C. (2010). Aggregatibacter actinomycetemcomitans leukotoxin: From threat to therapy. Journal of Dental Research, 89(6), 561–570. https://doi.org/10.1177/0022034510363682
Kachlany, S. C., Fine, D. H., & Figurski, D. H. (2000). Secretion of RTX leukotoxin by Actinobacillus actinomycetemcomitans. Infection and Immunity, 68(11), 6094–6100. https://doi.org/10.1128/IAI.68.11.6094‐6100.2000
Koessler, J., Niklaus, M., Weber, K., Koessler, A., Kuhn, S., Boeck, M., & Kobsar, A. (2019). The Role of human platelet preparation for toll‐like receptors 2 and 4 related platelet responsiveness. TH Open, 03(2), e94–e102. https://doi.org/10.1055/s‐0039‐1685495
Kozarov, E. V., Dorn, B. R., Shelburne, C. E., Dunn, W. A. Jr., & Progulske‐Fox, A. (2005). Human atherosclerotic plaque contains viable invasive Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. Arteriosclerosis, Thrombosis, and Vascular Biology, 25(3), e17–18. https://doi.org/10.1161/01.ATV.0000155018.67835.1a
Kraig, E., Dailey, T., & Kolodrubetz, D. (1990). Nucleotide sequence of the leukotoxin gene from Actinobacillus actinomycetemcomitans: homology to the alpha‐hemolysin/leukotoxin gene family. Infection and Immunity, 58(4), 920–929. https://doi.org/10.1128/iai.58.4.920‐929.1990
Krueger, E., & Brown, A. C. (2020). Aggregatibacter actinomycetemcomitans leukotoxin: From mechanism to targeted anti‐toxin therapeutics. Mol Oral Microbiol, 35(3), 85–105. https://doi.org/10.1111/omi.12284
Laky, M., Assinger, A., Esfandeyari, A., Bertl, K., Haririan, H., & Volf, I. (2011). Decreased phosphorylation of platelet vasodilator‐stimulated phosphoprotein in periodontitis–a role of periodontal pathogens. Thrombosis Research, 128(2), 155–160. https://doi.org/10.1016/j.thromres.2011.02.016
Lally, E. T., Golub, E. E., & Kieba, I. R. (1994). Identification and immunological characterization of the domain of Actinobacillus actinomycetemcomitans leukotoxin that determines its specificity for human target cells. Journal of Biological Chemistry, 269(49), 31289–31295. https://doi.org/10.1016/S0021‐9258(18)47421‐2
Lang, N. P., Joss, A., Orsanic, T., Gusberti, F. A., & Siegrist, B. E. (1986). Bleeding on probing—A predictor for the progression of periodontal‐disease. Journal of Clinical Periodontology, 13(6), 590–596. https://doi.org/10.1111/j.1600‐051X.1986.tb00852.x
McNicol, A. (2015). Bacteria‐induced intracellular signalling in platelets. Platelets, 26(4), 309–316. https://doi.org/10.3109/09537104.2015.1014470
Mills, K., Kelly, P., Tiraby, JG., Lioux, T., & Perouzel, E. (2016). Patent of novel compound, International Publication Number: WO 2016/008946 A1; World Intellectual Property Organization.
Moll, G. N., Konings, W. N., & Driessen, A. J. (1999). Bacteriocins: mechanism of membrane insertion and pore formation. Antonie Van Leeuwenhoek, 76(1–4), 185–198. https://doi.org/10.1023/A:1002002718501
Muller, H. P. (2009). Multilevel modeling of gingival bleeding on probing in young adult carriers of non‐JP2‐like strains of Aggregatibacter actinomycetemcomitans. Clinical Oral Investigations, 13(2), 171–178. https://doi.org/10.1007/s00784‐008‐0218‐4
Nakamori, M., Hosomi, N., Nishi, H., Aoki, S., Nezu, T., Shiga, Y., Kinoshita, N., Ishikawa, K., Imamura, E., Shintani, T., Ohge, H., Kawaguchi, H., Kurihara, H., Wakabayashi, S., & Maruyama, H. (2020). Serum IgG titers against periodontal pathogens are associated with cerebral hemorrhage growth and 3‐month outcome. PLoS ONE, 15(10), e0241205. https://doi.org/10.1371/journal.pone.0241205
Nichols, F. C., Bajrami, B., Clark, R. B., Housley, W., & Yao, X. (2012). Free lipid A isolated from Porphyromonas gingivalis lipopolysaccharide is contaminated with phosphorylated dihydroceramide lipids: recovery in diseased dental samples. Infection and Immunity, 80(2), 860–874. https://doi.org/10.1128/IAI.06180‐11
Nicu, E. A., Van der Velden, U., Nieuwland, R., Everts, V., & Loos, B. G. (2009). Elevated platelet and leukocyte response to oral bacteria in periodontitis. Journal of Thrombosis and Haemostasis, 7(1), 162–170. https://doi.org/10.1111/j.1538‐7836.2008.03219.x
Niklaus, M., Klingler, P., Weber, K., Koessler, A., Kuhn, S., Boeck, M., Kobsar, A., & Koessler, J. (2022). Platelet toll‐like‐receptor‐2 and ‐4 mediate different immune‐related responses to bacterial ligands. TH Open, 06(3), e156–e167. https://doi.org/10.1055/a‐1827‐7365
Park, E., Na, H. S., Song, Y. R., Shin, S. Y., Kim, Y. M., & Chung, J. (2014). Activation of NLRP3 and AIM2 inflammasomes by Porphyromonas gingivalis infection. Infection and Immunity, 82(1), 112–123. https://doi.org/10.1128/IAI.00862‐13
Pashenkov, M. V., Murugina, N. E., Budikhina, A. S., & Pinegin, B. V. (2019). Synergistic interactions between NOD receptors and TLRs: Mechanisms and clinical implications. Journal of Leukocyte Biology, 105(4), 669–680. https://doi.org/10.1002/JLB.2RU0718‐290R
Perrella, G., Nagy, M., Watson, S. P., & Heemskerk, J. W. M. (2021). Platelet GPVI (glycoprotein VI) and thrombotic complications in the venous system. Arteriosclerosis, Thrombosis, and Vascular Biology, 41(11), 2681–2692. https://doi.org/10.1161/ATVBAHA.121.316108
Pussinen, P. J., Alfthan, G., Jousilahti, P., Paju, S., & Tuomilehto, J. (2007). Systemic exposure to Porphyromonas gingivalis predicts incident stroke. Atherosclerosis, 193(1), 222–228. https://doi.org/10.1016/j.atherosclerosis.2006.06.027
Schwarz, U. R., Geiger, J., Walter, U., & Eigenthaler, M. (1999). Flow cytometry analysis of intracellular VASP phosphorylation for the assessment of activating and inhibitory signal transduction pathways in human platelets–definition and detection of ticlopidine/clopidogrel effects. Thromb Haemost, 82(3), 1145–1152. https://doi.org/10.1055/s‐0037‐1614344
Schwarz, U. R., Walter, U., & Eigenthaler, M. (2001). Taming platelets with cyclic nucleotides. Biochemical Pharmacology, 62(9), 1153–1161. https://doi.org/10.1016/s0006‐2952(01)00760‐2
Senini, V., Amara, U., Paul, M., & Kim, H. (2019). Porphyromonas gingivalis lipopolysaccharide activates platelet Cdc42 and promotes platelet spreading and thrombosis. Journal of Periodontology, 90(11), 1336–1345. https://doi.org/10.1002/JPER.18‐0596
Shattil, S. J., Hoxie, J. A., Cunningham, M., & Brass, L. F. (1985). Changes in the platelet membrane glycoprotein IIb·IIIa complex during platelet activation. Journal of Biological Chemistry, 260(20), 11107–11114. https://doi.org/10.1016/S0021‐9258(17)39154‐8
Uehara, A., Fujimoto, Y., Kawasaki, A., Kusumoto, S., Fukase, K., & Takada, H. (2006). Meso‐diaminopimelic acid and meso‐lanthionine, amino acids specific to bacterial peptidoglycans, activate human epithelial cells through NOD1. Journal of Immunology, 177(3), 1796–1804. https://doi.org/10.4049/jimmunol.177.3.1796
Yu, K. M., Inoue, Y., Umeda, M., Terasaki, H., Chen, Z. Y., & Iwai, T. (2011). The periodontal anaerobe Porphyromonas gingivalis induced platelet activation and increased aggregation in whole blood by rat model. Thrombosis Research, 127(5), 418–425. https://doi.org/10.1016/j.thromres.2010.12.004