Abstract
BackgroundLongicin is a defensin-like peptide, identified from the midgut epithelium of hard tick Haemaphysalis longicornis. Several studies have already shown the antimicrobial and parasiticidal activities of longicin peptide and one of its synthetic partial analogs, longicin P4. In this study, longicin peptides were tested for potential antiviral activity against Langat virus (LGTV), a tick-borne flavivirus.MethodsLongicin P1 and P4 peptides were chemically synthesized. Antiviral activity of the longicin peptides against LGTV was evaluated through in vitro virucidal assays, wherein the antiviral efficacy was determined by reduction in number of viral foci and virus yield. Additionally, longicin P4 was also tested for its activity against human adenovirus, a non-enveloped virus. Lastly, to assess the importance of longicin on the innate antiviral immunity of H. longicornis ticks, gene silencing through RNAi was performed.ResultsLongicin P4 produced significant viral foci reduction and lower virus yield against LGTV, while longicin P1 failed to demonstrate the same results. Conversely, both longicin partial analogs (P1 and P4) did not show significant antiviral activity when tested on adenovirus. In addition, longicin-silenced ticks showed significantly higher virus titer after 7 days post-infection but a significantly lower titer was detected after an additional 14 days of observation as compared to the Luc dsRNA-injected ticks. Mortality in both groups did not show any significant difference.ConclusionOur results suggest that longicin P4 has in vitro antiviral activity against LGTV but not against a non-enveloped virus such as adenovirus. Likewise, though most cationic antimicrobial peptides like longicin act directly on target membranes, the exact mechanism of membrane targeting of longicin P4 in enveloped viruses, such as LGTV, requires further investigation. Lastly, while the in vitro virucidal capacity of longicin P4 was confirmed in this study, the role of the endogenous tick longicin in the antiviral defense of H. longicornis against LGTV still remains to be demonstrated.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1344-5) contains supplementary material, which is available to authorized users.
Highlights
Longicin is a defensin-like peptide, identified from the midgut epithelium of hard tick Haemaphysalis longicornis
We investigated the virucidal activity of longicin P4 against Langat virus (LGTV), a member of tick-borne encephalitis virus (TBEV) serocomplex of the Flaviviridae family
Cytotoxicity activity of the longicin P4 peptide To eliminate the possibility that foci reduction was due to reduction in the number of viable host cells, we examined the cell growth inhibitory effect of partial peptides longicin P1 and P4 on BHK-21 cells (Fig. 1)
Summary
Longicin is a defensin-like peptide, identified from the midgut epithelium of hard tick Haemaphysalis longicornis. Antimicrobial peptides are ancient immune molecules that are important in invertebrate and vertebrate host defenses [8, 9] These peptides display broad-spectrum biological activity against bacteria, yeast, fungi, protozoan parasites and enveloped viruses [10,11,12] and have been demonstrated to possess immunomodulatory properties [13]. Numerous small molecules such as defensins, lysozymes or by tick-specific antimicrobial compounds such as microplusin provide the direct antimicrobial defense in ticks [2]
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