Korean J Ophthalmol.  2013 Jun;27(3):199-203. 10.3341/kjo.2013.27.3.199.

Anti-adenoviral Effects of Human Cationic Antimicrobial Protein-18/LL-37, an Antimicrobial Peptide, by Quantitative Polymerase Chain Reaction

• Uchio E
• Inoue H
• Kadonosono K

• KMID: 1798058
• DOI: http://doi.org/10.3341/kjo.2013.27.3.199

Abstract

PURPOSE
Antimicrobial peptides have an important role in self-protection of the ocular surface. Human cationic antimicrobial protein (hCAP)-18 is a linear, alpha-helical peptide that consists of a conserved pro-sequence called a cathelin-like domain and a C-terminal peptide named LL-37. We investigated the in vitro anti-adenoviral activity of hCAP-18/LL-37 in several adenovirus types, inducing keratoconjunctivitis.
METHODS
A549 cells were used for viral cell culture, and human adenovirus (HAdV) types 3 (HAdV3, species B), 4 (species E), 8, 19a, and 37 (species D) were used. The cytotoxicity of LL-37 was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay to obtain 50% cytotoxic concentration. After pretreatment of A549 cells with serial dilutions of LL-37 for 24 hours, adenovirus was cultured for seven days, and adenoviral DNA was quantitatively measured by real-time polymerase chain reaction (PCR).
RESULTS
The 50% effective concentration of LL-37 obtained by real-time PCR ranged between 118 and 270 microM. LL-37 showed a significant inhibitory effect on adenoviral proliferation in all adenovirus types except HAdV4 in a dose-dependent manner.
CONCLUSIONS
LL-37 has significant inhibitory activity against HAdV3, 8, and 19, which induce keratoconjunctivitis. These results indicate that hCAP-18/LL-37 may be a possible candidate for the treatment of HAdV keratoconjunctivitis.

Keyword

Adenoviridae; Antiviral agents; Human cationic antimicrobial protein-18; LL-37; Viral conjunctivitis

MeSH Terms

Adenocarcinoma
Adenoviridae/*drug effects/*genetics
Adenoviridae Infections/*drug therapy/virology
Antimicrobial Cationic Peptides/*pharmacology
Cell Line, Tumor
DNA, Viral/genetics
Humans
Keratoconjunctivitis/*drug therapy/virology
Lung Neoplasms
Reverse Transcriptase Polymerase Chain Reaction/methods
Antimicrobial Cationic Peptides
DNA, Viral

Figure

• 

  Fig. 1 Toxicity assay results of LL-37 in the A549 cell line.

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