Salivary Levels of Antibacterial Peptide (LL-37/hCAP-18) and Cotinine in Patients With Chronic Periodontitis
Corresponding Author
Yasuo Takeuchi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Correspondence: Yasuo Takeuchi, Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, 113-8549 Tokyo, Japan. Fax: 81-3-5803-0196; e-mail: [email protected].Search for more papers by this authorToshiyuki Nagasawa
Section of Periodontology and Endodontology, Health Sciences University of Hokkaido, Hokkaido, Japan.
Search for more papers by this authorSayaka Katagiri
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorSatoshi Kitagawara
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorHiroaki Kobayashi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorTatsurou Koyanagi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorYuichi Izumi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Global Center of Excellence Program for Tooth and Bone Research, Tokyo Medical and Dental University.
Search for more papers by this authorCorresponding Author
Yasuo Takeuchi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Correspondence: Yasuo Takeuchi, Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, 113-8549 Tokyo, Japan. Fax: 81-3-5803-0196; e-mail: [email protected].Search for more papers by this authorToshiyuki Nagasawa
Section of Periodontology and Endodontology, Health Sciences University of Hokkaido, Hokkaido, Japan.
Search for more papers by this authorSayaka Katagiri
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorSatoshi Kitagawara
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorHiroaki Kobayashi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorTatsurou Koyanagi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Search for more papers by this authorYuichi Izumi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Global Center of Excellence Program for Tooth and Bone Research, Tokyo Medical and Dental University.
Search for more papers by this authorAbstract
Background: The purpose of this study is to examine the relationship between salivary LL-37 levels and clinical severity in patients with chronic periodontitis (CP). The presence/absence of four periodontopathic bacteria and salivary cotinine levels were also examined to assess the impact of these factors on LL-37 production.
Methods: Unstimulated salivary samples were collected from 69 patients with CP. Salivary concentrations of LL-37 and cotinine were measured by enzyme-linked immunosorbent assay. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola in saliva were detected by polymerase chain reaction. Periodontal examination included determination of probing depth (PD), clinical attachment level, bleeding on probing, and plaque control record.
Results: Mean salivary LL-37 concentration was 225.0 ± 227.2 ng/mL, and a high prevalence of periodontopathic bacteria was observed. The stepwise ordinal logistic regression model showed that high salivary LL-37 levels were significantly associated with the presence of T. denticola and higher percentage of teeth with PD ≥5 mm. In addition, higher salivary cotinine levels (≥8 ng/mL) were negatively associated with salivary LL-37 levels.
Conclusions: Salivary LL-37 level was positively correlated with severe periodontal destruction, and production was apparently associated with periodontopathic bacterial infection. The negative correlations between salivary LL-37 and cotinine levels also suggest that smoking or long-term exposure to environmental tobacco smoke can lead to lower LL-37 levels in the oral cavity and increased risk of periodontitis.
REFERENCES
- 1Gudmundsson GH, Agerberth B. Neutrophil antibacterial peptides, multifunctional effector molecules in the mammalian immune system. J Immunol Methods 1999; 232: 45–54.
- 2Diamond G, Beckloff N, Ryan LK. Host defense peptides in the oral cavity and the lung: Similarities and differences. J Dent Res 2008; 87: 915–927.
- 3Guthmiller JM, Vargas KG, Srikantha R, et al. Susceptibilities of oral bacteria and yeast to mammalian cathelicidins. Antimicrob Agents Chemother 2001; 45: 3216–3219.
- 4Ouhara K, Komatsuzawa H, Yamada S, et al. Susceptibilities of periodontopathogenic and cariogenic bacteria to antibacterial peptides, β-defensins and LL37, produced by human epithelial cells. J Antimicrob Chemother 2005; 55: 888–896.
- 5Altman H, Steinberg D, Porat Y, et al. In vitro assessment of antimicrobial peptides as potential agents against several oral bacteria. J Antimicrob Chemother 2006; 58: 198–201.
- 6Braff MH, Hawkins MA, Di Nardo A, et al. Structure-function relationships among human cathelicidin peptides: Dissociation of antimicrobial properties from host immunostimulatory activities. J Immunol 2005; 174: 4271–4278.
- 7Lee SH, Jun HK, Lee HR, Chung CP, Choi BK. Antibacterial and lipopolysaccharide (LPS)-neutralising activity of human cationic antimicrobial peptides against periodontopathogens. Int J Antimicrob Agents 2010; 35: 138–145.
- 8Inomata M, Into T, Murakami Y. Suppressive effect of the antimicrobial peptide LL-37 on expression of IL-6, IL-8 and CXCL10 induced by Porphyromonas gingivalis cells and extracts in human gingival fibroblasts. Eur J Oral Sci 2010; 118: 574–581.
- 9Mookherjee N, Brown KL, Bowdish DM, et al. Modulation of the TLR-mediated inflammatory response by the endogenous human host defense peptide LL-37. J Immunol 2006; 176: 2455–2464.
- 10Nagaoka I, Tamura H, Hirata M. An antimicrobial cathelicidin peptide, human CAP18/LL-37, suppresses neutrophil apoptosis via the activation of formyl-peptide receptor-like 1 and P2X7. J Immunol 2006; 176: 3044–3052.
- 11Barlow PG, Li Y, Wilkinson TS, et al. The human cationic host defense peptide LL-37 mediates contrasting effects on apoptotic pathways in different primary cells of the innate immune system. J Leukoc Biol 2006; 80: 509–520.
- 12Zheng Y, Niyonsaba F, Ushio H, et al. Cathelicidin LL-37 induces the generation of reactive oxygen species and release of human alpha-defensins from neutrophils. Br J Dermatol 2007; 157: 1124–1131.
- 13De Yang, Chen Q, Schmidt AP, et al. LL-37, the neutrophil granule- and epithelial cell-derived cathelicidin, utilizes formyl peptide receptor-like 1 (FPRL1) as a receptor to chemoattract human peripheral blood neutrophils, monocytes, and T cells. J Exp Med 2000; 192: 1069–1074.
- 14Chertov O, Michiel DF, Xu L, et al. Identification of defensin-1, defensin-2, and CAP37/azurocidin as T-cell chemoattractant proteins released from interleukin-8-stimulated neutrophils. J Biol Chem 1996; 271: 2935–2940.
- 15Nagaoka I, Hirota S, Niyonsaba F, et al. Cathelicidin family of antibacterial peptides CAP18 and CAP11 inhibit the expression of TNF-alpha by blocking the binding of LPS to CD14(+) cells. J Immunol 2001; 167: 3329–3338.
- 16Koczulla R, von Degenfeld G, Kupatt C, et al. An angiogenic role for the human peptide antibiotic LL-37/hCAP-18. J Clin Invest 2003; 111: 1665–1672.
- 17Pütsep K, Carlsson G, Boman HG, Andersson M. Deficiency of antibacterial peptides in patients with morbus Kostmann: An observation study. Lancet 2002; 360: 1144–1149.
- 18de Haar SF, Jansen DC, Schoenmaker T, De Vree H, Everts V, Beertsen W. Loss-of-function mutations in cathepsin C in two families with Papillon-Lefèvre syndrome are associated with deficiency of serine proteinases in PMNs. Hum Mutat 2004; 23: 524.
- 19Karlsson J, Carlsson G, Ramme KG, et al. Low plasma levels of the protein pro-LL-37 as an early indication of severe disease in patients with chronic neutropenia. Br J Haematol 2007; 137: 166–169.
- 20Türkoğlu O, Emingil G, Kütükçüler N, Atilla G. Gingival crevicular fluid levels of cathelicidin LL-37 and interleukin-18 in patients with chronic periodontitis. J Periodontol 2009; 80: 969–976.
- 21Puklo M, Guentsch A, Hiemstra PS, Eick S, Potempa J. Analysis of neutrophil-derived antimicrobial peptides in gingival crevicular fluid suggests importance of cathelicidin LL-37 in the innate immune response against periodontogenic bacteria. Oral Microbiol Immunol 2008; 23: 328–335.
- 22Yamamoto Y, Nishida N, Tanaka M, et al. Association between passive and active smoking evaluated by salivary cotinine and periodontitis. J Clin Periodontol 2005; 32: 1041–1046.
- 23Sayers NM, James JA, Drucker DB, Blinkhorn AS. Possible potentiation of toxins from Prevotella intermedia, Prevotella nigrescens, and Porphyromonas gingivalis by cotinine. J Periodontol 1999; 70: 1269–1275.
- 24Flemmig TF. Periodontitis. Ann Periodontol 1999; 4: 32–38.
- 25O'Leary TJ, Drake RB, Naylor JE. The plaque control record. J Periodontol 1972; 43: 38.
- 26Ashimoto A, Chen C, Bakker I, Slots J. Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions. Oral Microbiol Immunol 1996; 11: 266–273.
- 27Overhage J, Campisano A, Bains M, Torfs EC, Rehm BH, Hancock RE. Human host defense peptide LL-37 prevents bacterial biofilm formation. Infect Immun 2008; 76: 4176–4182.
- 28de Haar SF, Hiemstra PS, van Steenbergen MT, Everts V, Beertsen W. Role of polymorphonuclear leukocyte-derived serine proteinases in defense against Actinobacillus actinomycetemcomitans. Infect Immun 2006; 74: 5284–5291.
- 29Sanz M, Lau L, Herrera D, Morillo JM, Silva A. Methods of detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis in periodontal microbiology, with special emphasis on advanced molecular techniques: A review. J Clin Periodontol 2004; 31: 1034–1047.
- 30Hosokawa I, Hosokawa Y, Komatsuzawa H, et al. Innate immune peptide LL-37 displays distinct expression pattern from beta-defensins in inflamed gingival tissue. Clin Exp Immunol 2006; 146: 218–225.
- 31Kinane DF, Chestnutt IG. Smoking and periodontal disease. Crit Rev Oral Biol Med 2000; 11: 356–365.
- 32MacFarlane GD, Herzberg MC, Wolff LF, Hardie NA. Refractory periodontitis associated with abnormal polymorphonuclear leukocyte phagocytosis and cigarette smoking. J Periodontol 1992; 63: 908–913.
- 33Persson L, Bergström J, Ito H, Gustafsson A. Tobacco smoking and neutrophil activity in patients with periodontal disease. J Periodontol 2001; 72: 90–95.
- 34Giannopoulou C, Roehrich N, Mombelli A. Effect of nicotine-treated epithelial cells on the proliferation and collagen production of gingival fibroblasts. J Clin Periodontol 2001; 28: 769–775.
- 35Gutner M, Chaushu S, Balter D, Bachrach G. Saliva enables the antimicrobial activity of LL-37 in the presence of proteases of Porphyromonas gingivalis. Infect Immun 2009; 77: 5558–5563.
- 36Ji S, Hyun J, Park E, Lee BL, Kim KK, Choi Y. Susceptibility of various oral bacteria to antimicrobial peptides and to phagocytosis by neutrophils. J Periodontal Res 2007; 42: 410–419.
- 37Scott MG, Davidson DJ, Gold MR, Bowdish D, Hancock RE. The human antimicrobial peptide LL-37 is a multifunctional modulator of innate immune responses. J Immunol 2002; 169: 3883–3891.
- 38Fukumoto K, Nagaoka I, Yamataka A, et al. Effect of antibacterial cathelicidin peptide CAP18/LL-37 on sepsis in neonatal rats. Pediatr Surg Int 2005; 21: 20–24.
- 39Alalwani SM, Sierigk J, Herr C, et al. The antimicrobial peptide LL-37 modulates the inflammatory and host defense response of human neutrophils. Eur J Immunol 2010; 40: 1118–1126.
