title: Enhancement of antiviral activity of human alpha-defensin 5 against herpes simplex virus 2 by arginine mutagenesis at adaptive evolution sites.
abstract: Herpes simplex virus 2 (HSV-2) infection is still one of the common causes of sexually transmitted diseases worldwide. The prevalence of HSV strains resistant to traditional nucleoside antiviral agents has led to the development of novel antiviral drugs. Human alpha-defensin 5 (HD5), a kind of endogenous antimicrobial peptide expressed in the epithelia of the small intestine and urogenital tract, displays natural antiviral activity. Based on arginine-rich features and adaptive evolution characteristics of vertebrate defensins, we conducted a screen for HD5 derivatives with enhanced anti-HSV-2 activity by a single arginine substitution at the adaptive evolution sites. Cell protection assay and temporal antiviral studies showed that HD5 and its mutants displayed affirmatory but differential anti-HSV-2 effects in vitro by inhibiting viral adhesion and entry. Inspiringly, the E21R-HD5 mutant had significantly higher antiviral activity than natural HD5, which is possibly attributed to the stronger binding affinity of the E21R-HD5 mutant with HSV-2 capsid protein gD, indicating that E21R mutation can increase the anti-HSV-2 potency of HD5. In a mouse model of lethal HSV-2 infection, prophylactic and/or therapeutic treatment with E21R-HD5 via intravaginal instillation remarkably alleviated the symptoms and delayed disease progress and resulted in about a 1.5-fold-higher survival rate than in the HD5 group. Furthermore, the E21R variant exhibited a 2-fold-higher antiviral potency against HIV-1 over parental HD5 in vitro. This study demonstrates that arginine mutagenesis at appropriate evolution sites may significantly enhance the antiviral activity of HD5, which also paves a facile way to search for potent antiviral drugs based on natural antimicrobial peptides.
authors.affiliation: Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing, China.
authors.forename: Aiping
authors.initials: A
authors.lastname: Wang
authors.forename: Fang
authors.initials: F
authors.lastname: Chen
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Wang
authors.forename: Mingqiang
authors.initials: M
authors.lastname: Shen
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.forename: Jian
authors.initials: J
authors.lastname: Hu
authors.forename: Song
authors.initials: S
authors.lastname: Wang
authors.forename: Fang
authors.initials: F
authors.lastname: Geng
authors.forename: Cheng
authors.initials: C
authors.lastname: Wang
authors.forename: Xinze
authors.initials: X
authors.lastname: Ran
authors.forename: Yongping
authors.initials: Y
authors.lastname: Su
authors.forename: Tianmin
authors.initials: T
authors.lastname: Cheng
authors.forename: Junping
authors.initials: J
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology*; D036022:Capsid Proteins / Q000187:drug effects / Q000378:metabolism; D002522:Chlorocebus aethiops; D019143:Evolution, Molecular; D005260:Female; D015658:HIV Infections / Q000188:drug therapy* / Q000517:prevention & control; D015497:HIV-1 / Q000187:drug effects; D006561:Herpes Simplex / Q000188:drug therapy* / Q000517:prevention & control; D018258:Herpesvirus 2, Human / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D016296:Mutagenesis; D009154:Mutation; D016415:Sequence Alignment; D014709:Vero Cells; D019562:Viral Load; D053585:Virus Attachment / Q000187:drug effects*; D023084:alpha-Defensins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D019380:Anti-HIV Agents; D000998:Antiviral Agents; D036022:Capsid Proteins; C500225:DEFA5 protein, human; D023084:alpha-Defensins