Cranberry Proanthocyanidins Neutralize the Effects of Aggregatibacter actinomycetemcomitans Leukotoxin
Abstract
:1. Introduction
2. Material and Methods
2.1. Cranberry PACs Isolation and Characterization
2.2. Effect of Cranberry PACs on Leukotoxin Gene (ltxA, ltxB, ltxC, and ltxD) Expression
2.3. LtxA Purification
2.4. Cell Culture
2.5. Real-Time Cell Viability
2.6. Cell Membrane Permeability Assay
2.7. Identification of Apoptotic Cells by Annexin Staining and Determination of Caspase-1 Activation
2.8. Caspase-1 Quantification and Cytokine Analysis
2.9. P2X7 and CIAS Gene Expression
2.10. Determination of Intracellular Reactive Oxygen Species (ROS)
2.11. LtxA Binding Assay
2.12. Statistical Analysis
3. Results
3.1. Effect of Cranberry PACs on the Expression of Leukotoxin Genes and Secretion of LtxA
3.2. Effect of Cranberry PACs on Leukotoxin Activity
3.3. Identification of Apoptotic Cells by Annexin Staining and Determination of Caspase 1 Activation
3.4. Caspase-1 Quantification and Cytokine Analysis
3.5. P2X7 and CIAS Gene Expression
3.6. Measurement of Intracellular ROS and Superoxide
3.7. Binding of FITC–LtxA to Macrophages
4. Discussion
Author Contributions
Funding
Conflicts of Interest
References
- Henderson, B.; Ward, J.M.; Ready, D. Aggregatibacter (Actinobacillus) actinomycetemcomitans: A triple A* periodontopathogen? Periodontol. 2000 2010, 54, 78–105. [Google Scholar] [CrossRef] [PubMed]
- Albandar, J.M. Aggressive and acute periodontal diseases. Periodontol. 2000 2014, 65, 7–12. [Google Scholar] [CrossRef] [PubMed]
- Susin, C.; Haas, A.N.; Albandar, J.M. Epidemiology and demographics of aggressive periodontitis. Periodontol. 2000 2014, 65, 27–45. [Google Scholar] [CrossRef]
- Hoglund Aberg, C.; Kwamin, F.; Claesson, R.; Dahlen, G.; Johansson, A.; Haubek, D. Progression of attachment loss is strongly associated with presence of the JP2 genotype of Aggregatibacter actinomycetemcomitans: A prospective cohort study of a young adolescent population. J. Clin. Periodontol. 2014, 41, 232–241. [Google Scholar] [CrossRef]
- Fine, D.H.; Markowitz, K.; Furgang, D.; Fairlie, K.; Ferrandiz, J.; Nasri, C.; McKiernan, M.; Gunsolley, J. Aggregatibacter actinomycetemcomitans and its relationship to initiation of localized aggressive periodontitis: Longitudinal cohort study of initially healthy adolescents. J. Clin. Microbiol. 2007, 45, 3859–3869. [Google Scholar] [CrossRef]
- Haubek, D.; Ennibi, O.K.; Poulsen, K.; Vaeth, M.; Poulsen, S.; Kilian, M. Risk of aggressive periodontitis in adolescent carriers of the JP2 clone of Aggregatibacter (Actinobacillus) actinomycetemcomitans in Morocco: A prospective longitudinal cohort study. Lancet 2008, 371, 237–242. [Google Scholar] [CrossRef]
- Aberg, C.H.; Sjodin, B.; Lakio, L.; Pussinen, P.J.; Johansson, A.; Claesson, R. Presence of Aggregatibacter actinomycetemcomitans in young individuals: A 16-year clinical and microbiological follow-up study. J. Clin. Periodontol. 2009, 36, 815–822. [Google Scholar] [CrossRef]
- Aberg, C.H.; Kelk, P.; Johansson, A. Aggregatibacter actinomycetemcomitans: Virulence of its leukotoxin and association with aggressive periodontitis. Virulence 2015, 6, 188–195. [Google Scholar] [CrossRef]
- Dahlen, G.; Claesson, R.; Aberg, C.H.; Haubek, D.; Johansson, A.; Kwamin, F. Subgingival bacteria in Ghanaian adolescents with or without progression of attachment loss. J. Oral Microbiol. 2014, 6. [Google Scholar] [CrossRef]
- Johansson, A. Aggregatibacter actinomycetemcomitans leukotoxin: A powerful tool with capacity to cause imbalance in the host inflammatory response. Toxins 2011, 3, 242–259. [Google Scholar] [CrossRef]
- Kieselbach, T.; Zijnge, V.; Granstrom, E.; Oscarsson, J. Proteomics of Aggregatibacter actinomycetemcomitans outer membrane vesicles. PLoS ONE 2015, 10, e0138591. [Google Scholar] [CrossRef] [PubMed]
- Hajishengallis, G. The inflammophilic character of the periodontitis-associated microbiota. Mol. Oral Microbiol. 2014, 29, 248–257. [Google Scholar] [CrossRef] [PubMed]
- Meyle, J.; Chapple, I. Molecular aspects of the pathogenesis of periodontitis. Periodontol. 2000 2015, 69, 7–17. [Google Scholar] [CrossRef] [PubMed]
- Kelk, P.; Abd, H.; Claesson, R.; Sandstrom, G.; Sjostedt, A.; Johansson, A. Cellular and molecular response of human macrophages exposed to Aggregatibacter actinomycetemcomitans leukotoxin. Cell Death Dis. 2011, 2, e126. [Google Scholar] [CrossRef] [PubMed]
- Taabazuing, C.Y.; Okondo, M.C.; Bachovchin, D.A. Pyroptosis and apoptosis pathways engage in bidirectional crosstalk in monocytes and macrophages. Cell Chem. Biol. 2017, 24, 507–514. [Google Scholar] [CrossRef] [PubMed]
- Fink, S.L.; Cookson, B.T. Caspase-1-dependent pore formation during pyroptosis leads to osmotic lysis of infected host macrophages. Cell. Microbiol. 2006, 8, 1812–1825. [Google Scholar] [CrossRef]
- Kelk, P.; Johansson, A.; Claesson, R.; Hanstrom, L.; Kalfas, S. Caspase 1 involvement in human monocyte lysis induced by Actinobacillus actinomycetemcomitans leukotoxin. Infect. Immun. 2003, 71, 4448–4455. [Google Scholar] [CrossRef]
- Kelk, P.; Claesson, R.; Hanstrom, L.; Lerner, U.H.; Kalfas, S.; Johansson, A. Abundant secretion of bioactive interleukin-1beta by human macrophages induced by Actinobacillus actinomycetemcomitans leukotoxin. Infect. Immun. 2005, 73, 453–458. [Google Scholar] [CrossRef]
- Jorgensen, I.; Rayamajhi, M.; Miao, E.A. Programmed cell death as a defence against infection. Nat. Rev. Immunol. 2017, 17, 151–164. [Google Scholar] [CrossRef]
- Kaye, E.K. Nutrition, dietary guidelines and optimal periodontal health. Periodontol. 2000 2012, 58, 93–111. [Google Scholar] [CrossRef]
- Bodet, C.; Piche, M.; Chandad, F.; Grenier, D. Inhibition of periodontopathogen-derived proteolytic enzymes by a high-molecular-weight fraction isolated from cranberry. J. Antimicrob. Chemother. 2006, 57, 685–690. [Google Scholar] [CrossRef] [PubMed]
- Bodet, C.; Chandad, F.; Grenier, D. Cranberry components inhibit interleukin-6, interleukin-8, and prostaglandin E production by lipopolysaccharide-activated gingival fibroblasts. Eur. J. Oral Sci. 2007, 115, 64–70. [Google Scholar] [CrossRef] [PubMed]
- Bodet, C.; Chandad, F.; Grenier, D. Inhibition of host extracellular matrix destructive enzyme production and activity by a high-molecular-weight cranberry fraction. J. Periodontal Res. 2007, 42, 159–168. [Google Scholar] [CrossRef]
- Feghali, K.; Feldman, M.; La, V.D.; Santos, J.; Grenier, D. Cranberry proanthocyanidins: Natural weapons against periodontal diseases. J. Agric. Food Chem. 2012, 60, 5728–5735. [Google Scholar] [CrossRef]
- La, V.D.; Labrecque, J.; Grenier, D. Cytoprotective effect of proanthocyanidin-rich cranberry fraction against bacterial cell wall-mediated toxicity in macrophages and epithelial cells. Phytother. Res. 2009, 23, 1449–1452. [Google Scholar] [CrossRef]
- Foo, L.Y.; Lu, Y.; Howell, A.B.; Vorsa, N. A-Type proanthocyanidin trimers from cranberry that inhibit adherence of uropathogenic P-fimbriated Escherichia coli. J. Nat. Prod. 2000, 63, 1225–1228. [Google Scholar] [CrossRef]
- Foo, L.Y.; Lu, Y.; Howell, A.B.; Vorsa, N. The structure of cranberry proanthocyanidins which inhibit adherence of uropathogenic P-fimbriated Escherichia coli in vitro. Phytochemistry 2000, 54, 173–181. [Google Scholar] [CrossRef]
- Isaza, M.P.; Duncan, M.S.; Kaplan, J.B.; Kachlany, S.C. Screen for leukotoxin mutants in Aggregatibacter actinomycetemcomitans: Genes of the phosphotransferase system are required for leukotoxin biosynthesis. Infect. Immun. 2008, 76, 3561–3568. [Google Scholar] [CrossRef]
- Tsai, C.C.; Shenker, B.J.; DiRienzo, J.M.; Malamud, D.; Taichman, N.S. Extraction and isolation of a leukotoxin from Actinobacillus actinomycetemcomitans with polymyxin B. Infect. Immun. 1984, 43, 700–705. [Google Scholar]
- Fine, D.H.; Furgang, D.; Schreiner, H.C.; Goncharoff, P.; Charlesworth, J.; Ghazwan, G.; Fitzgerald-Bocarsly, P.; Figurski, D.H. Phenotypic variation in Actinobacillus actinomycetemcomitans during laboratory growth: Implications for virulence. Microbiology 1999, 145 Pt 6, 1335–1347. [Google Scholar] [CrossRef]
- Kachlany, S.C.; Fine, D.H.; Figurski, D.H. Purification of secreted leukotoxin (LtxA) from Actinobacillus actinomycetemcomitans. Protein Expr. Purif. 2002, 25, 465–471. [Google Scholar] [CrossRef]
- Rovera, G.; Santoli, D.; Damsky, C. Human promyelocytic leukemia cells in culture differentiate into macrophage-like cells when treated with a phorbol diester. Proc. Natl. Acad. Sci. USA 1979, 76, 2779–2783. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Feldman, M.; La, V.D.; Lombardo Bedran, T.B.; Palomari Spolidorio, D.M.; Grenier, D. Porphyromonas gingivalis-mediated shedding of extracellular matrix metalloproteinase inducer (EMMPRIN) by oral epithelial cells: A potential role in inflammatory periodontal disease. Microbes Infect. 2011, 13, 1261–1269. [Google Scholar] [CrossRef] [PubMed]
- Kittichotirat, W.; Bumgarner, R.E.; Asikainen, S.; Chen, C. Identification of the pangenome and its components in 14 distinct Aggregatibacter actinomycetemcomitans strains by comparative genomic analysis. PLoS ONE 2011, 6, e22420. [Google Scholar] [CrossRef] [PubMed]
- Kittichotirat, W.; Bumgarner, R.E.; Chen, C. Evolutionary divergence of Aggregatibacter actinomycetemcomitans. J. Dent. Res. 2016, 95, 94–101. [Google Scholar] [CrossRef] [Green Version]
- Brogan, J.M.; Lally, E.T.; Poulsen, K.; Kilian, M.; Demuth, D.R. Regulation of Actinobacillus actinomycetemcomitans leukotoxin expression: Analysis of the promoter regions of leukotoxic and minimally leukotoxic strains. Infect. Immun. 1994, 62, 501–508. [Google Scholar]
- Guthmiller, J.M.; Lally, E.T.; Korostoff, J. Beyond the specific plaque hypothesis: Are highly leukotoxic strains of Actinobacillus actinomycetemcomitans a paradigm for periodontal pathogenesis? Crit. Rev. Oral Biol. Med. 2001, 12, 116–124. [Google Scholar] [CrossRef]
- Johansson, A.; Claesson, R.; Hanstrom, L.; Sandstrom, G.; Kalfas, S. Polymorphonuclear leukocyte degranulation induced by leukotoxin from Actinobacillus actinomycetemcomitans. J. Periodontal Res. 2000, 35, 85–92. [Google Scholar] [CrossRef]
- Saxen, L.; Asikainen, S. Metronidazole in the treatment of localized juvenile periodontitis. J. Clin. Periodontol. 1993, 20, 166–171. [Google Scholar] [CrossRef]
- Walker, C.B. The acquisition of antibiotic resistance in the periodontal microflora. Periodontol. 2000 1996, 10, 79–88. [Google Scholar] [CrossRef]
- Deas, D.E.; Mealey, B.L. Response of chronic and aggressive periodontitis to treatment. Periodontol. 2000 2010, 53, 154–166. [Google Scholar] [CrossRef] [PubMed]
- Mombelli, A.; Gmur, R.; Gobbi, C.; Lang, N.P. Actinobacillus actinomycetemcomitans in adult periodontitis. II. Characterization of isolated strains and effect of mechanical periodontal treatment. J. Periodontol. 1994, 65, 827–834. [Google Scholar] [CrossRef] [PubMed]
- Kachlany, S.C. Aggregatibacter actinomycetemcomitans leukotoxin: From threat to therapy. J. Dent. Res. 2010, 89, 561–570. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Crosby, J.A.; Kachlany, S.C. TdeA, a TolC-like protein required for toxin and drug export in Aggregatibacter (Actinobacillus) actinomycetemcomitans. Gene 2007, 388, 83–92. [Google Scholar] [CrossRef] [Green Version]
- Balashova, N.V.; Shah, C.; Patel, J.K.; Megalla, S.; Kachlany, S.C. Aggregatibacter actinomycetemcomitans LtxC is required for leukotoxin activity and initial interaction between toxin and host cells. Gene 2009, 443, 42–47. [Google Scholar] [CrossRef]
- Fong, K.P.; Pacheco, C.M.; Otis, L.L.; Baranwal, S.; Kieba, I.R.; Harrison, G.; Hersh, E.V.; Boesze-Battaglia, K.; Lally, E.T. Actinobacillus actinomycetemcomitans leukotoxin requires lipid microdomains for target cell cytotoxicity. Cell. Microbiol. 2006, 8, 1753–1767. [Google Scholar] [CrossRef] [Green Version]
- Korostoff, J.; Wang, J.F.; Kieba, I.; Miller, M.; Shenker, B.J.; Lally, E.T. Actinobacillus actinomycetemcomitans leukotoxin induces apoptosis in HL-60 cells. Infect. Immun. 1998, 66, 4474–4483. [Google Scholar]
- Mangan, D.F.; Taichman, N.S.; Lally, E.T.; Wahl, S.M. Lethal effects of Actinobacillus actinomycetemcomitans leukotoxin on human T lymphocytes. Infect. Immun. 1991, 59, 3267–3272. [Google Scholar]
- Nalbant, A.; Chen, C.; Wang, Y.; Zadeh, H.H. Induction of T-cell apoptosis by Actinobacillus actinomycetemcomitans mutants with deletion of ltxA and cdtABC genes: Possible activity of GroEL-like molecule. Oral Microbiol. Immunol. 2003, 18, 339–349. [Google Scholar] [CrossRef]
- Kwamin, F.; Gref, R.; Haubek, D.; Johansson, A. Interactions of extracts from selected chewing stick sources with Aggregatibacter actinomycetemcomitans. BMC Res. Notes 2012, 5, 203. [Google Scholar] [CrossRef] [Green Version]
- Diaz-de-Cerio, E.; Pasini, F.; Verardo, V.; Fernandez-Gutierrez, A.; Segura-Carretero, A.; Caboni, M.F. Psidium guajava L. leaves as source of proanthocyanidins: Optimization of the extraction method by RSM and study of the degree of polymerization by NP-HPLC-FLD-ESI-MS. J. Pharm. Biomed. Anal. 2017, 133, 1–7. [Google Scholar] [CrossRef] [PubMed]
- Miller, D.K.; Ayala, J.M.; Egger, L.A.; Raju, S.M.; Yamin, T.T.; Ding, G.J.; Gaffney, E.P.; Howard, A.D.; Palyha, O.C.; Rolando, A.M.; et al. Purification and characterization of active human interleukin-1 beta-converting enzyme from THP.1 monocytic cells. J. Biol. Chem. 1993, 268, 18062–18069. [Google Scholar] [PubMed]
- Li, P.; Allen, H.; Banerjee, S.; Franklin, S.; Herzog, L.; Johnston, C.; McDowell, J.; Paskind, M.; Rodman, L.; Salfeld, J.; et al. Mice deficient in IL-1 beta-converting enzyme are defective in production of mature IL-1 beta and resistant to endotoxic shock. Cell 1995, 80, 401–411. [Google Scholar] [CrossRef] [Green Version]
- Martinon, F.; Burns, K.; Tschopp, J. The inflammasome: A molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol. Cell 2002, 10, 417–426. [Google Scholar] [CrossRef]
- Ogura, Y.; Sutterwala, F.S.; Flavell, R.A. The inflammasome: First line of the immune response to cell stress. Cell 2006, 126, 659–662. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bloemen, V.; Schoenmaker, T.; de Vries, T.J.; Everts, V. IL-1beta favors osteoclastogenesis via supporting human periodontal ligament fibroblasts. J. Cell. Biochem. 2011, 112, 1890–1897. [Google Scholar] [CrossRef]
- Delaleu, N.; Bickel, M. Interleukin-1 beta and interleukin-18: Regulation and activity in local inflammation. Periodontol. 2000 2004, 35, 42–52. [Google Scholar] [CrossRef]
- Kanneganti, T.D.; Ozoren, N.; Body-Malapel, M.; Amer, A.; Park, J.H.; Franchi, L.; Whitfield, J.; Barchet, W.; Colonna, M.; Vandenabeele, P.; et al. Bacterial RNA and small antiviral compounds activate caspase-1 through cryopyrin/Nalp3. Nature 2006, 440, 233–236. [Google Scholar] [CrossRef] [Green Version]
- Mariathasan, S.; Weiss, D.S.; Newton, K.; McBride, J.; O’Rourke, K.; Roose-Girma, M.; Lee, W.P.; Weinrauch, Y.; Monack, D.M.; Dixit, V.M. Cryopyrin activates the inflammasome in response to toxins and ATP. Nature 2006, 440, 228–232. [Google Scholar] [CrossRef]
- Martinon, F.; Petrilli, V.; Mayor, A.; Tardivel, A.; Tschopp, J. Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature 2006, 440, 237–241. [Google Scholar] [CrossRef] [Green Version]
- Skaper, S.D.; Debetto, P.; Giusti, P. The P2X7 purinergic receptor: From physiology to neurological disorders. FASEB J. 2010, 24, 337–345. [Google Scholar] [CrossRef] [PubMed]
- Ferrari, D.; Pizzirani, C.; Adinolfi, E.; Lemoli, R.M.; Curti, A.; Idzko, M.; Panther, E.; Di Virgilio, F. The P2X7 receptor: A key player in IL-1 processing and release. J. Immunol. 2006, 176, 3877–3883. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Solini, A.; Menini, S.; Rossi, C.; Ricci, C.; Santini, E.; Blasetti Fantauzzi, C.; Iacobini, C.; Pugliese, G. The purinergic 2X7 receptor participates in renal inflammation and injury induced by high-fat diet: Possible role of NLRP3 inflammasome activation. J. Pathol. 2013, 231, 342–353. [Google Scholar] [CrossRef] [PubMed]
- Harijith, A.; Ebenezer, D.L.; Natarajan, V. Reactive oxygen species at the crossroads of inflammasome and inflammation. Front. Physiol. 2014, 5, 352. [Google Scholar] [CrossRef]
- Fukumoto, J.; Fukumoto, I.; Parthasarathy, P.T.; Cox, R.; Huynh, B.; Ramanathan, G.K.; Venugopal, R.B.; Allen-Gipson, D.S.; Lockey, R.F.; Kolliputi, N. NLRP3 deletion protects from hyperoxia-induced acute lung injury. Am. J. Physiol. Cell Physiol. 2013, 305, C182–C189. [Google Scholar] [CrossRef]
- Heid, M.E.; Keyel, P.A.; Kamga, C.; Shiva, S.; Watkins, S.C.; Salter, R.D. Mitochondrial reactive oxygen species induces NLRP3-dependent lysosomal damage and inflammasome activation. J. Immunol. 2013, 191, 5230–5238. [Google Scholar] [CrossRef] [Green Version]
- Krueger, E.; Hayes, S.; Chang, E.H.; Yutuc, S.; Brown, A.C. Receptor-Based Peptides for Inhibition of Leukotoxin Activity. ACS Infect. Dis. 2018, 4, 1073–1081. [Google Scholar] [CrossRef]
- Stabholz, A.; Taichman, N.S.; Soskolne, W.A. Occurrence of Actinobacillus actinomycetemcomitans and anti-leukotoxin antibodies in some members of an extended family affected by Papillon-Lefevre syndrome. J. Periodontol. 1995, 66, 653–657. [Google Scholar] [CrossRef]
- Kleinfelder, J.W.; Topoll, H.H.; Preus, H.R.; Muller, R.F.; Lange, D.E.; Bocker, W. Microbiological and immunohistological findings in a patient with Papillon-Lefevre syndrome. J. Clin. Periodontol. 1996, 23, 1032–1038. [Google Scholar] [CrossRef]
Gene | Primer Sequence |
---|---|
GAPDH | Sense: 5′-GGTATCGTGGAAGGACTCATGAC-3′ Antisense: 5′-ATGCCAGTGAGCTTCCCGTTCAGC-3′ |
CIAS | Sense: 5′-CATTAAGATGGAGTTGCTGTTTGAC-3′ Antisense: 5′-CCGACAGTGGATATAGAACAGATAG-3 |
P2X7 | Sense: 5′-GAAACGGACTCTGATAAAAGTCTTC-3′ Antisense: 5′-TCTTCCTGTAGTAGTATTCGTTGAC-3′ |
16S rRNA | Sense: 5′-CCTGAATAATGTGGTGATAGTG-3′ Antisense: 5′-CCTCTCTCTATGAACAAGAACG-3′ |
ltxA | Sense: 5′-GTGCTAGGTAAACATCGGTAAAG-3′ Antisense: 5′-GACCACAGAGGCAATTAACC-3′ |
ltxB | Sense: 5′-CTTAGATATCAGTCAGGGAGAAG-3′ Antisense: 5′-CTCTCTGATACTTCGATTAAGCAAC-3′ |
ltxC | Sense: 5′-CATCTCTTGTTTATGACGACTG-3′ Antisense: 5′-GTTTATCGACTTTACCTCCATG-3′ |
ltxD | Sense: 5′-CCAGCAAGTCTCTGAAATTCG-3′ Antisense: 5′-CTTCTTCCGGCACAACTACC-3 |
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Ben Lagha, A.; Howell, A.; Grenier, D. Cranberry Proanthocyanidins Neutralize the Effects of Aggregatibacter actinomycetemcomitans Leukotoxin. Toxins 2019, 11, 662. https://doi.org/10.3390/toxins11110662
Ben Lagha A, Howell A, Grenier D. Cranberry Proanthocyanidins Neutralize the Effects of Aggregatibacter actinomycetemcomitans Leukotoxin. Toxins. 2019; 11(11):662. https://doi.org/10.3390/toxins11110662
Chicago/Turabian StyleBen Lagha, Amel, Amy Howell, and Daniel Grenier. 2019. "Cranberry Proanthocyanidins Neutralize the Effects of Aggregatibacter actinomycetemcomitans Leukotoxin" Toxins 11, no. 11: 662. https://doi.org/10.3390/toxins11110662