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The antimicrobial peptide HBD-2 and the Toll-like receptors-2 and -4 are induced in synovial membranes in case of septic arthritis

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Abstract

Septic arthritis is frequently observed especially in immune-compromised or chronically diseased patients and leads to functional impairment due to tissue destruction. Recently, production of antimicrobial peptides (AMP) was observed in articular cartilage after exposure to bacteria. This report examines the role of synoviocyte-derived AMPs in innate defense mechanisms of articular joints. Samples of healthy, low-grade synovialitis and septic synovial membranes were assessed for the expression of human β-defensin-2 (HBD-2) and Toll-like receptor-2 and -4 (TLR) by immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). A stable synoviocyte line (K4IM) was used for in vitro experiments and assayed for endogenous HBD-2 and TLR production after exposure to inflammatory cytokines or bacterial supernatants by reverse transcription polymerase chain reaction (RT-PCR), real-time RT-PCR, Western blot, ELISA, and dual luciferase assay. Healthy human synovial membranes and cultured synoviocytes are able to produce HBD-2 and TLR-1–5 at basal expression levels. Samples of bacteria-colonized synovial membranes produce higher levels of HBD-2 when compared with samples of healthy tissues. K4IM synoviocytes exposed to Staphylococcus aureus, Pseudomonas aeruginosa, or proinflammatory cytokines demonstrated a clear HBD-2 transcription and protein induction. TLR-2 and -4 are known to have a critical role in the recognition of gram-positive and gram-negative bacteria in epithelia and are induced in mesenchymal synoviocytes after bacterial exposure on transcription and on protein level. This report demonstrates an unappreciated role of synovial membranes: samples of septic synovial membranes and cultured synoviocytes exposed to bacteria produce increased amounts of the AMP HBD-2 and the bacteria recognition receptors TLR-2 and -4. The induction of anti-inflammatory pathways in infected synoviocytes suggests involvement in intra-articular defense mechanisms.

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Acknowledgments

We wish to thank Susanne Echterhagen, Christiane Jaeschke, Patrycja Kozak, Inka Kronenbitter, Ursula Mundt, Michaela Nicolau, Angela Rüben, Sonja Seiter, and Kirsten Vosen for their expert technical assistance and Clemens Franke for the drawing. The K4IM and the HSE cells were a courtesy from Christian Kaps. This work was supported by grants of the Deutsche Forschungsgemeinschaft (SFB 617, project A22; DFG Va 220/2-1, Pu 214/3-2, Pu 214/4-2, Pu 214/5-2, and PA 738/9-1), from the “Stiftung zur Förderung der Medizinischen Forschung” of the Medical Faculty of the University of Kiel (to D V and TP), from the Hensel Stiftung (to DV and FP), and by the “Verein zur Förderung der Erforschung und Bekämpfung rheumatischer Erkrankungen Bad Bramstedt e.V.”, and by BMBF/Wilhelm-Roux (FKZ 9/16 to FP).

Conflicts of interest

We declare that we have no conflict of interest.

Authors’ contributions

DV, TP, EK, CW, LB, FP, MT, and SL performed the experiments, and BT and AS contributed to the draft manuscript; DV and TP contributed equally to the present work. The manuscript has been read and approved by all authors.

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Authors and Affiliations

  1. Department of Orthopaedic Trauma Surgery, UKSH Campus Kiel, Kiel, Germany

    D. Varoga, S. Lippross & A. Seekamp

  2. Department of Anatomy, Christian-Albrechts-University, Kiel, Germany

    D. Varoga, E. Klostermeier, C. Wruck, L. O. Brandenburg, B. Tillmann & T. Pufe

  3. Institute of Anatomy and Cell Biology, Martin-Luther-University of Halle-Wittenberg, Halle, Germany

    F. Paulsen

  4. Department of Anatomy and Cell Biology, RWTH Aachen University, Wendlingweg 2, 52074, Aachen, Germany

    C. Wruck, L. O. Brandenburg, M. Tohidnezhad & T. Pufe

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  1. D. Varoga
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  2. E. Klostermeier
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  3. F. Paulsen
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  4. C. Wruck
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  7. M. Tohidnezhad
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  8. A. Seekamp
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  9. B. Tillmann
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  10. T. Pufe
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Corresponding author

Correspondence to T. Pufe.

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The experiments comply with the current laws of Germany.

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Varoga, D., Klostermeier, E., Paulsen, F. et al. The antimicrobial peptide HBD-2 and the Toll-like receptors-2 and -4 are induced in synovial membranes in case of septic arthritis. Virchows Arch 454, 685–694 (2009). https://doi.org/10.1007/s00428-009-0780-4

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  • DOI: https://doi.org/10.1007/s00428-009-0780-4

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