PMID- 31095741
OWN - NLM
STAT- MEDLINE
DCOM- 20200115
LR  - 20221207
IS  - 1600-0765 (Electronic)
IS  - 0022-3484 (Linking)
VI  - 54
IP  - 6
DP  - 2019 Dec
TI  - The host defense peptide LL-37 is internalized by human periodontal ligament 
      cells and prevents LPS-induced MCP-1 production.
PG  - 662-670
LID - 10.1111/jre.12667 [doi]
AB  - OBJECTIVE: The human host defense peptide LL-37 both shows antimicrobial effects 
      and modulates host cell properties. Here, we assess the effects of synthesized 
      LL-37 on lipopolysaccharide (LPS)-induced inflammation in human periodontal 
      ligament (PDL) cells and investigates underlying mechanisms. BACKGROUND: LL-37 
      has been detected in the periodontal tissues, but its functional importance for 
      PDL cell innate immune responses is not known. METHODS: Human PDL cells were 
      obtained from premolars extracted on orthodontic indications. Cellular 
      pro-inflammatory monocyte chemoattractant protein-1 (MCP-1) mRNA expression was 
      determined using quantitative real-time RT-PCR. MCP-1 protein production was 
      assessed by western blot and ELISA. Internalization of LL-37 by PDL cells was 
      visualized by immunocytochemistry. Nuclear factor kappa-light-chain-enhancer of 
      activated B-cell (NF-kappaB) activity was assessed by western blot of phosphorylated 
      p65, phosphorylated p105, and IkappaBalpha proteins. Binding of LL-37 to PDL cell DNA was 
      determined by isolation and purification of DNA and dot blot for LL-37 
      immunoreactivity. RESULTS: Treatment with LL-37 (1 micromol/L) for 24 hours prevented 
      LPS-induced stimulation of MCP-1 expression analyzed both on transcript and on 
      protein levels. Stimulation with LL-37 (1 micromol/L) for 24 hours had no effect on 
      toll-like receptor (TLR)2 and TLR4 transcript expression, suggesting that LL-37 
      acts downstream of the TLRs. Preincubation with LL-37 for 60 minutes followed by 
      stimulation with LPS for 24 hours in the absence of LL-37 completely prevented 
      LPS-evoked MCP-1 transcript expression, implying that LL-37 acts intracellularly 
      and not via binding and neutralization of LPS. In PDL cells stimulated with LL-37 
      for 60 minutes, the peptide was internalized as demonstrated by 
      immunocytochemistry, suggesting an intracellular mechanism of action. LL-37 
      immunoreactivity was observed both in the cytosol and in the nucleus. 
      Downregulation of LPS-induced MCP-1 by LL-37 was not mediated by reduction in 
      NF-kappaB activity as shown by unaltered expression of phosphorylated p65, 
      phosphorylated p105, and IkappaBalpha NF-kappaB proteins in the presence of LL-37. 
      Immunoreactivity for LL-37 was observed in PDL cell DNA treated with but not 
      without 0.1 and 1 micromol/L LL-37 for 60 minutes in vitro. CONCLUSION: LL-37 
      abolishes LPS-induced MCP-1 production in human PDL cells through an 
      intracellular, NF-kappaB-independent mechanism which probably involves direct 
      interaction between LL-37 and DNA.
CI  - (c) 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
FAU - Aidoukovitch, Alexandra
AU  - Aidoukovitch A
AD  - Department of Experimental Medical Science, Lund University, Lund, Sweden.
AD  - Folktandvarden Skane, Lund, Sweden.
FAU - Anders, Emma
AU  - Anders E
AD  - Department of Experimental Medical Science, Lund University, Lund, Sweden.
FAU - Dahl, Sara
AU  - Dahl S
AD  - Department of Experimental Medical Science, Lund University, Lund, Sweden.
FAU - Nebel, Daniel
AU  - Nebel D
AD  - Department of Experimental Medical Science, Lund University, Lund, Sweden.
FAU - Svensson, Daniel
AU  - Svensson D
AD  - Department of Experimental Medical Science, Lund University, Lund, Sweden.
AD  - Department of Women's and Children's Health, Karolinska Institute, Solna, Sweden.
FAU - Nilsson, Bengt-Olof
AU  - Nilsson BO
AUID- ORCID: 0000-0003-2337-8412
AD  - Department of Experimental Medical Science, Lund University, Lund, Sweden.
LA  - eng
GR  - 20171116/The Royal Physiographic Society/
GR  - 20171031/The Swedish Dental Society/
GR  - 20180764/The Crafoord Foundation/
GR  - OFRS 732821/The Research Funds for Oral Health Related Research by Region Skane/
GR  - Medical Faculty, Lund University/
PT  - Journal Article
DEP - 20190516
PL  - United States
TA  - J Periodontal Res
JT  - Journal of periodontal research
JID - 0055107
RN  - 0 (Antimicrobial Cationic Peptides)
RN  - 0 (CCL2 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (NF-kappa B)
RN  - 0 (Cathelicidins)
SB  - IM
MH  - Antimicrobial Cationic Peptides/*pharmacology
MH  - Chemokine CCL2/*metabolism
MH  - Humans
MH  - Lipopolysaccharides
MH  - NF-kappa B/metabolism
MH  - Periodontal Ligament/*cytology/drug effects
MH  - Cathelicidins
OTO - NOTNLM
OT  - NF-kappaB
OT  - antimicrobial peptide
OT  - inflammation
OT  - innate immunity
EDAT- 2019/05/17 06:00
MHDA- 2020/01/16 06:00
CRDT- 2019/05/17 06:00
PHST- 2018/12/14 00:00 [received]
PHST- 2019/03/15 00:00 [revised]
PHST- 2019/04/20 00:00 [accepted]
PHST- 2019/05/17 06:00 [pubmed]
PHST- 2020/01/16 06:00 [medline]
PHST- 2019/05/17 06:00 [entrez]
AID - 10.1111/jre.12667 [doi]
PST - ppublish
SO  - J Periodontal Res. 2019 Dec;54(6):662-670. doi: 10.1111/jre.12667. Epub 2019 May 
      16.