Defensins are broad-spectrum antimicrobial peptides that are components of innate immunity. To date, only epithelial surfaces and blood cells have been shown to produce these cationic peptides in bactericidal concentrations when challenged with microorganisms or inflammatory cytokines. Infections caused by gram-negative pathogens occur only infrequently in association with joint surgery. The present study was undertaken to investigate whether this may be explained by intraarticular production of gram-negative–specialized antimicrobial peptides.

Methods

Healthy articular cartilage and cultured T/C-28a2 chondrocytes were assessed, by reverse transcriptase–polymerase chain reaction (RT-PCR) and immunohistochemistry, for expression of various antimicrobial peptides. The expression of human β-defensin 2 (HBD-2) was studied in cultured chondrocytes after exposure to bacterial supernatants and proinflammatory cytokines and was assayed by real-time RT-PCR and immunoblot analysis. A septic arthritis mouse model was used to investigate the regulation of the murine homolog of HBD-2 in articular cartilage after bacterial inoculation.

Results

Healthy articular cartilage and T/C-28a2 chondrocytes were able to produce different antimicrobial peptides. After exposure to gram-negative bacteria and proinflammatory cytokines, expression of cartilage-derived HBD-2 strongly increased. Immunoblot analysis revealed up-regulation of the gram-negative–specialized HBD-2 in microbicidal doses. Immunohistochemistry analysis revealed induction of the murine homolog of HBD-2 in vivo after intraarticular injection of bacteria.

Conclusion

This study demonstrated a previously unrecognized function of human chondrocytes. In addition to its biomechanical properties, articular cartilage has the ability to produce antimicrobial substances when challenged with microorganisms. The expression of HBD-2 in microbicidal doses suggests that antimicrobial peptides may contribute to host defense mechanisms in articular joints.

REFERENCES

1 Hultgren OH, Stenson M, Tarkowski A. Role of IL-12 in Staphylococcus aureus-triggered arthritis and sepsis. Arthritis Res 2001; 3: 41–7.
10.1186/ar138
CAS PubMed Web of Science® Google Scholar
2 Ryan MJ, Kavanagh R, Wall PG, Hazleman BL. Bacterial joint infections in England and Wales: analysis of bacterial isolates over a four year period. Br J Rheumatol 1997; 36: 370–3.
10.1093/rheumatology/36.3.370
CAS PubMed Web of Science® Google Scholar
3 Conforti L, Petrovic M, Mohammad D, Lee S, Ma Q, Barone S, et al. Hypoxia regulates expression and activity of Kv1.3 channels in T lymphocytes: a possible role in T cell proliferation. J Immunol 2003; 170: 695–702.
10.4049/jimmunol.170.2.695
CAS PubMed Web of Science® Google Scholar
4 Harder J, Bartels J, Christophers E, Schroder JM. A peptide antibiotic from human skin [letter]. Nature 1997; 387: 861.
10.1038/43088
CAS PubMed Web of Science® Google Scholar
5 Dorschner RA, Lin KH, Murakami M, Gallo RL. Neonatal skin in mice and humans expresses increased levels of antimicrobial peptides: innate immunity during development of the adaptive response. Pediatr Res 2003; 53: 566–72.
10.1203/01.PDR.0000057205.64451.B7
CAS PubMed Web of Science® Google Scholar
6 Schroder JM, Harder J. Human β-defensin-2. Int J Biochem Cell Biol 1999; 31: 645–51.
10.1016/S1357-2725(99)00013-8
CAS PubMed Web of Science® Google Scholar
7 Singh PK, Jia HP, Wiles K, Hesselberth J, Liu L, Conway BA, et al. Production of β-defensins by human airway epithelia. Proc Natl Acad Sci U S A 1998; 95: 14961–6.
10.1073/pnas.95.25.14961
CAS PubMed Web of Science® Google Scholar
8 Harder J, Meyer-Hoffert U, Teran LM, Schwichtenberg L, Bartels J, Maune S, et al. Mucoid Pseudomonas aeruginosa, TNF-α, and IL-1-β, but not IL-6, induce human β-defensin-2 in respiratory epithelia. Am J Respir Cell Mol Biol 2000; 22: 714–21.
10.1165/ajrcmb.22.6.4023
CAS PubMed Web of Science® Google Scholar
9 Zasloff M. Antimicrobial peptides of multicellular organisms. Nature 2002; 415: 389–95.
10.1038/415389a
CAS PubMed Web of Science® Google Scholar
10 Tsutsumi-Ishii Y, Nagaoka I. NF-κB-mediated transcriptional regulation of human β-defensin-2 gene following lipopolysaccharide stimulation. J Leukoc Biol 2002; 71: 154–62.
10.1189/jlb.71.1.154
CAS PubMed Web of Science® Google Scholar
11 Bensch KW, Raida M, Magert HJ, Schulz-Knappe P, Forssmann WG. HBD-1: a novel β-defensin from human plasma. FEBS Lett 1995; 17: 368: 331–5.
10.1016/0014-5793(95)00687-5
Web of Science® Google Scholar
12 Harder J, Bartels J, Christophers E, Schroder JM. Isolation and characterization of human β-defensin-3, a novel human inducible peptide antibiotic. J Biol Chem 2001; 276: 5707–13.
10.1074/jbc.M008557200
CAS PubMed Web of Science® Google Scholar
13 Garcia JR, Krause A, Schulz S, Rodriguez-Jimenez FJ, Kluver E, Adermann K, et al. Human β-defensin 4: a novel inducible peptide with a specific salt-sensitive spectrum of antimicrobial activity. FASEB J 2001; 15: 1819–21.
10.1096/fj.00-0865fje
CAS PubMed Web of Science® Google Scholar
14 Yang D, Chertov O, Bykovskaia SN, Chen Q, Buffo MJ, Shogan J, et al. β-defensins: linking innate and adaptive immunity through dendritic and T cell CCR6. Science 1999; 286: 525–8.
10.1126/science.286.5439.525
CAS PubMed Web of Science® Google Scholar
15 Sorensen OE, Cowland JB, Theilgaard-Monch K, Liu L, Ganz T, Borregaard N. Wound healing and expression of antimicrobial peptides/polypeptides in human keratinocytes, a consequence of common growth factors. J Immunol 2003; 170: 5583–9.
10.4049/jimmunol.170.11.5583
CAS PubMed Web of Science® Google Scholar
16 Koczulla R, von Degenfeld G, Kupatt C, Krotz F, Zahler S, Gloe T, et al. An angiogenic role for the human peptide antibiotic LL-37/hCAP-18. J Clin Invest 2003; 11: 1665–72.
10.1172/JCI17545
Web of Science® Google Scholar
17 Bals R, Wang X, Meegalla RL, Wattler S, Weiner DJ, Nehls MC, et al. Mouse β-defensin 3 is an inducible antimicrobial peptide expressed in the epithelia of multiple organs. Infect Immun 1999; 67: 3542–7.
10.1128/IAI.67.7.3542-3547.1999
CAS PubMed Web of Science® Google Scholar
18 Yamaguchi Y, Nagase T, Makita R, Fukuhara S, Tomita T, Tominaga T, et al. Identification of multiple novel epididymis-specific β-defensin isoforms in humans and mice. J Immunol 2002; 169: 2516–23.
10.4049/jimmunol.169.5.2516
CAS PubMed Web of Science® Google Scholar
19 Morrison GM, Davidson DJ, Dorin JR. A novel mouse βa defensin, Defb2, which is upregulated in the airways by lipopolysaccharide. FEBS Lett 1999; 442: 112–6.
10.1016/S0014-5793(98)01630-5
CAS PubMed Web of Science® Google Scholar
20 Jia HP, Wowk SA, Schutte BC, Lee SK, Vivado A, Tack BF, et al. A novel murine β-defensin expressed in tongue, esophagus, and trachea. J Biol Chem 2000; 275: 33314–20.
10.1074/jbc.M006603200
CAS PubMed Web of Science® Google Scholar
21 Burd RS, Furrer JL, Sullivan J, Smith AL. Murine β-defensin-3 is an inducible peptide with limited tissue expression and broad-spectrum antimicrobial activity. Shock 2002; 18: 461–4.
10.1097/00024382-200211000-00013
PubMed Web of Science® Google Scholar
22 Paulsen F, Pufe T, Petersen W, Tillmann B. Expression of natural peptide antibiotics in human articular cartilage and synovial membrane. Clin Diagn Lab Immunol 2001; 8: 1021–3.
10.1128/CDLI.8.5.1021-1023.2001
CAS PubMed Web of Science® Google Scholar
23 Paulsen F, Pufe T, Conradi L, Varoga D, Tsokos M, Papendieck J, et al. Antimicrobial peptides are expressed and produced in healthy and inflamed human synovial membranes. J Pathol 2002; 198: 369–77.
10.1002/path.1224
CAS PubMed Web of Science® Google Scholar
24 Goldring MB, Birkhead JR, Suen LF, Yamin R, Mizuno S, Glowacki J, et al. Interleukin-1 β-modulated gene expression in immortalized human chondrocytes. J Clin Invest 1994; 94: 2307–16.
10.1172/JCI117595
CAS PubMed Web of Science® Google Scholar
25 Loeser R, Sadiev S, Tan L, Goldring MB. Integrin expression by primary and immortalized human chondrocytes: evidence of a differential role for α1β1 and α2β1 integrins mediating chondrocyte adhesion to types II and VI collagen. Osteoarthritis Cartilage 2000; 8: 96–105.
10.1053/joca.1999.0277
CAS PubMed Web of Science® Google Scholar
26 Osaki M, Tan L, Choy BK, Yoshida Y, Auron PE, Cheah KS, et al. The TATA-containing core promoter of the type II collagen gene (COL2A1) is the target of interferon-γ-mediated inhibition in human chondrocytes: requirement for Stat1α, Jak1, and Jak2. Biochem J 2003; 369: 103–15.
10.1042/bj20020928
CAS PubMed Web of Science® Google Scholar
27 Parker WL, Goldring MB, Philip A. Endoglin is expressed on human chondrocytes and forms a heteromeric complex with betaglycan in a ligand and type II TGF-β receptor independent manner. J Bone Miner Res 2003; 18: 289–302.
10.1359/jbmr.2003.18.2.289
CAS PubMed Web of Science® Google Scholar
28 Tan L, Peng H, Osaki M, Choy BK, Auron PE, Sandell LJ, et al. Egr-1 mediates transcriptional repression of COL2A1 promoter activity by interleukin-1β. J Biol Chem 2003; 278: 17688–700.
10.1074/jbc.M301676200
CAS PubMed Web of Science® Google Scholar
29 Grall F, Gu X, Tan L, Cho JY, Inan MS, Pettit A, et al. Responses to the proinflammatory cytokines interleukin-1 and tumor necrosis factor α in cells derived from rheumatoid synovium and other joint tissues involve nuclear factor κB–mediated induction of the Ets transcription factor ESE-1. Arthritis Rheum 2003; 48: 1249–60.
10.1002/art.10942
CAS PubMed Web of Science® Google Scholar
30 Deng GM, Nilsson IM, Verdrengh M, Tarkowski A. Intraarticularly localized bacterial DNA containing CpG motifs induces arthritis. Nat Med 1999; 5: 702–5.
10.1038/9554
CAS PubMed Web of Science® Google Scholar
31 Liu L, Roberts AA, Ganz T. By IL-1 signaling, monocyte-derived cells dramatically enhance the epidermal antimicrobial response to lipopolysaccharide. J Immunol 2003; 170: 575–80.
10.4049/jimmunol.170.1.575
CAS PubMed Web of Science® Google Scholar
32 Bals R, Wang X, Wu Z, Freeman T, Bafna V, Zasloff M, et al. Human β-defensin 2 is a salt-sensitive peptide antibiotic expressed in human lung. J Clin Invest 1998; 102: 874–80.
10.1172/JCI2410
CAS PubMed Web of Science® Google Scholar
33 Becker MN, Diamond G, Verghese MW, Randell SH. CD-14-dependent lipopolysaccharide-induced β-defensin-2 expression in human tracheobronchial epithelium. J Biol Chem 2000; 275: 29731–6.
10.1074/jbc.M000184200
CAS PubMed Web of Science® Google Scholar
34 Valore EV, Park CH, Quale AJ, Wiles KR, McCray PB, Ganz T. Human β-defensin-1: an antimicrobial peptide of urogenital tissue. J Clin Invest 1998; 101: 1633–42.
10.1172/JCI1861
CAS PubMed Web of Science® Google Scholar
35 O'Neil DA, Porter EM, Elewaut D, Anderson GM, Eckmann L, Ganz T, et al. Expression and regulation of the human β-defensins hBD-1 and hBD-2 in intestinal epithelium. J Immunol 1999; 163: 6718–24.
10.4049/jimmunol.163.12.6718
CAS PubMed Web of Science® Google Scholar
36 Spitznagel JK. Antibiotic proteins of human neutrophils. J Clin Invest 1990; 86: 1381–6.
10.1172/JCI114851
CAS PubMed Web of Science® Google Scholar
37 Harder J, Schroder JM. RNase-7, a novel innate immune defense antimicrobial protein of healthy human skin. J Biol Chem 2002; 277: 46779–84.
10.1074/jbc.M207587200
CAS PubMed Web of Science® Google Scholar
38 Ong PY, Ohtake T, Brandt C, Strickland I, Boguniewicz M, Ganz T, et al. Endogenous antimicrobial peptides and skin infections in atopic dermatitis. N Engl J Med 2002; 347: 1151–60.
10.1056/NEJMoa021481
CAS PubMed Web of Science® Google Scholar
39 Putsep K, Carlsson G, Boman HG, Andersson M. Deficiency of antibacterial peptides in patients with morbus Kostmann: an observation study. Lancet 2002; 360: 1116–7.
10.1016/S0140-6736(02)11201-3
PubMed Web of Science® Google Scholar
40 Salzman NH, Ghosh D, Huttner KM, Paterson Y, Bevins CL. Protection against enteric salmonellosis in transgenic mice expressing a human intestinal defensin. Nature 2003; 422: 522–6.
10.1038/nature01520
CAS PubMed Web of Science® Google Scholar

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Volume50, Issue11

November 2004

Pages 3526-3534

Production of endogenous antibiotics in articular cartilage

Abstract

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Production of endogenous antibiotics in articular cartilage

Abstract

Objective

Methods

Results

Conclusion

REFERENCES

Citing Literature

References

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