Antimicrobial peptides derived from the cartilage.-specific C-type Lectin Domain Family 3 Member A (CLEC3A) - potential in the prevention and treatment of septic arthritis

D Elezagic; M Mörgelin; G Hermes; A Hamprecht; G Sengle; D Lau; S Höllriegl; R Wagener; M Paulsson; T Streichert; A R Klatt

doi:10.1016/j.joca.2019.06.007

Antimicrobial peptides derived from the cartilage.-specific C-type Lectin Domain Family 3 Member A (CLEC3A) - potential in the prevention and treatment of septic arthritis

Osteoarthritis Cartilage. 2019 Oct;27(10):1564-1573. doi: 10.1016/j.joca.2019.06.007. Epub 2019 Jul 4.

Authors

D Elezagic¹, M Mörgelin², G Hermes¹, A Hamprecht³, G Sengle⁴, D Lau¹, S Höllriegl⁵, R Wagener⁶, M Paulsson⁷, T Streichert¹, A R Klatt⁸

Affiliations

¹ Institute for Clinical Chemistry, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany.
² Department of Clinical Sciences, Division of Infection Medicine, Biomedical Center, Lund University, 22184 Lund, Sweden.
³ Institute for Medical Microbiology, Immunology and Hygiene, Faculty of Medicine, University of Cologne, 50935 Cologne, Germany.
⁴ Center for Biochemistry, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany; Center for Molecular Medicine Cologne, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany; Department of Pediatrics and Adolescent Medicine, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.
⁵ Cologne Braunsfeld Trinity Hospital, 50933 Cologne, Germany.
⁶ Center for Biochemistry, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany; Center for Molecular Medicine Cologne, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany.
⁷ Center for Biochemistry, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany; Center for Molecular Medicine Cologne, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany; Cologne Excellence Cluster on Cellular Stress Responses in Ageing-associated Diseases, University of Cologne, 50931 Cologne, Germany.
⁸ Institute for Clinical Chemistry, Faculty of Medicine, University of Cologne, 50931 Cologne, Germany. Electronic address: andreas.klatt@uk-koeln.de.

PMID: 31279936
DOI: 10.1016/j.joca.2019.06.007

Abstract

Objective: To investigate the antimicrobial activity of peptides derived from C-type Lectin Domain Family 3 Member A (CLEC3A), shed light on the mechanism of antimicrobial activity and assess their potential application in prevention and treatment of septic arthritis.

Design: We performed immunoblot to detect CLEC3A peptides in human cartilage extracts. To investigate their antimicrobial activity, we designed peptides and recombinantly expressed CLEC3A domains and used them to perform viable count assays using E.coli, P.aeruginosa and S.aureus. We investigated the mechanism of their antimicrobial activity by fluorescence and scanning electron microscopy, performed ELISA-style immunoassays and transmission electron microscopy to test for lipopolysaccharide binding and surface plasmon resonance to test for lipoteichoic acid (LTA) binding. We coated CLEC3A peptides on titanium, a commonly used prosthetic material, and performed fluorescence microscopy to quantify bacterial adhesion. Moreover, we assessed the peptides' cytotoxicity against primary human chondrocytes using MTT cell viability assays.

Results: CLEC3A fragments were detected in human cartilage extracts. Moreover, bacterial supernatants lead to fragmentation of recombinant and cartilage-derived CLEC3A. CLEC3A-derived peptides killed E.coli, P.aeruginosa and S.aureus, permeabilized bacterial membranes and bound lipopolysaccharide and LTA. Coating CLEC3A antimicrobial peptides (AMPs) on titanium lead to significantly reduced bacterial adhesion to the material. In addition, microbicidal concentrations of CLEC3A peptides in vitro displayed no direct cytotoxicity against primary human chondrocytes.

Conclusions: We identify cartilage-specific AMPs originating from CLEC3A, resolve the mechanism of their antimicrobial activity and point to a novel approach in the prevention and treatment of septic arthritis using potent, non-toxic, AMPs.

Keywords: Antimicrobial peptides; Arthroplasty; Coating of prostheses; Septic arthritis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Anti-Bacterial Agents / pharmacology*
Anti-Bacterial Agents / therapeutic use*
Arthritis, Infectious / drug therapy*
Arthritis, Infectious / prevention & control*
Bacteria / drug effects*
Cartilage / metabolism
Humans
Lectins, C-Type* / metabolism
Peptides / metabolism
Peptides / therapeutic use*

Substances

Anti-Bacterial Agents
CLEC3A protein, human
Lectins, C-Type
Peptides