Abstract
Vibrio vulnificus causes fatal infections in humans, and antibiotics are commonly used in treatment regimens against V. vulnificus infection. However, the therapeutic effects of antibiotics are limited by multidrug resistance. In this study, we demonstrated that an antimicrobial peptide (AMP), HPA3PHis, loaded onto a gold nanoparticle-DNA aptamer (AuNP-Apt) conjugate (AuNP-Apt-HPA3PHis) is an effective therapeutic tool against V. vulnificus infection in vivo in mice. HPA3PHis induced bacterial cell death through the disruption of membrane integrity of V. vulnificus. The introduction of AuNP-Apt-HPA3PHis into V. vulnificus-infected HeLa cells dramatically reduced intracellular V. vulnificus by 90%, leading to an increase in the viability of the infected cells. Moreover, when V. vulnificus-infected mice were intravenously injected with AuNP-Apt-HPA3PHis, a complete inhibition of V. vulnificus colonization was observed in the mouse organs, leading to a 100% survival rate among the treated mice, whereas all the control mice died within 40 hours of being infected. Therefore, this study demonstrated the potential of an AMP delivered by AuNP-Apt as an effective and rapid treatment option against infection caused by a major pathogen in humans and aquatic animals.
Highlights
The emergence and spread of multidrug-resistant bacteria are global concerns
We investigated the effectiveness of a Helicobacter pylori-derived antimicrobial peptide (AMP), HPA3P, delivered by AuNP-Apt against V. vulnificus infection, which involved mechanisms of infection and virulence different from those of S. typhimurium
This His-tagging to HPA3P did not alter the minimal inhibitory concentrations (MICs) and minimal bactericidal activity (MBC) against V. vulnificus (Table 1), and AuNP-AptHis itself exhibited no bactericidal activity up to 10 nM
Summary
The emergence and spread of multidrug-resistant bacteria are global concerns. Overcoming antibiotic resistance is an urgent and imperative issue. HP(2-20), a 19-amino-acid peptide (AKKVFKRLEKLFSKIQNDK) derived from the N-terminus of Helicobacter pylori ribosomal protein L1 (RpL1), shows broad-spectrum antimicrobial activity against bacteria, fungi, and protozoa without causing hemolysis[14,15]. The instability of peptides in vivo and low penetrability in host cells are the major limiting factors for the clinical application of AMPs17. To overcome these limitations, we developed an AMP delivery system using gold nanoparticle-DNA aptamer (AuNP-Apt) conjugate. We investigated the effectiveness of a Helicobacter pylori-derived AMP, HPA3P, delivered by AuNP-Apt against V. vulnificus infection, which involved mechanisms of infection and virulence different from those of S. typhimurium. We found that the in vivo delivery of HPA3P loaded on AuNP-Apt effectively inhibited V. vulnificus, leading to 100% survival rate among V. vulnificus-infected mice
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