Cathelicidin antimicrobial peptides (human LL-37 and mouse
CRAMP) are mainly virucidal to enveloped virus. However, the effects and relative mechanisms of LL-37 and
CRAMP on non-enveloped virus are elusive. We herein found that
CRAMP expression was significantly up-regulated post non-enveloped Enterovirus 71 (EV71)
infection in different tissues of newborn ICR mice, while EV71 replication gradually declined post
CRAMP up-regulation, indicating the
antiviral potential of
cathelicidin against EV71. In vitro
antiviral assay showed that LL-37 and
CRAMP markedly reduced cytopathic effects (CPE), intracellular
viral RNA copy numbers, viral VP1
protein levels, and extracellular virons in U251 cells post EV71
infection, indicating that LL-37 and
CRAMP significantly inhibited EV71 replication. Mechanism of action assay showed that LL-37 and
CRAMP were not virucidal to EV71, but markedly regulated
antiviral immune response in U251 cells. Co-incubation of LL-37 or
CRAMP with U251 cells markedly increased the basal
interferon-β (IFN-β) expression and
interferon regulatory
transcription factor 3 (IRF3) phosphorylation, modestly enhanced IFN-β production and IRF3 phosphorylation upon EV71
infection, and significantly reduced
interleukin-6 (IL-6) production and
p38 mitogen-activated protein kinase (MAPK) activation post EV71
infection. Additionally, LL-37 and
CRAMP directly inhibited viral binding to U251 cells. Collectively, LL-37 and
CRAMP markedly inhibited EV71 replication via regulating
antiviral response and inhibiting viral binding, providing potent candidates for
peptide drug development against EV71
infection.