Design and preliminary study of VP1-encoding mRNA-lipid nanoparticles for Enterovirus 71
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Abstract
Background: Hand, foot, and mouth disease (HFMD) remains a major public health concern in the Asia-Pacific region, with Enterovirus 71 (EV71) being a major cause of severe neurological complications in children. The genetic diversity of EV71 highlights the need for adaptable vaccine platforms.
Objective: This study aimed to design and preliminarily evaluate an EV71 VP1-encoding mRNA-lipid nanoparticle (mRNA-LNP) construct as an antigen delivery approach for EV71 vaccine development.
Materials and methods: A codon-optimized VP1 mRNA was synthesized by in vitro transcription and formulated into lipid nanoparticles via microfluidic mixing. Physicochemical properties were characterized, and in vitro expression was assessed in FreeStyle 293F cells by western blot and immunofluorescence. VP1-specific humoral immune responses were evaluated in BALB/c mice following a prime-boost immunization regimen using ELISA.
Results: The mRNA-LNPs exhibited a mean particle size of 99.24±0.46 nm, a low polydispersity index (0.041±0.004), a slightly negative zeta potential (-20.54±0.98 mV), and an encapsulation efficiency of 92.56±1.53%. In vitro studies demonstrated VP1 protein expression following mRNA delivery. Immunization induced VP1-specific IgG responses, with antibody levels significantly increased after prime (1.254±0.775) and boost (4.225±0.322) compared with control groups (p<0.01 and p<0.001, respectively). Endpoint titers increased from a geometric mean titer of 2,000 (95% CI: 901.50-4,437.00) after priming to 256,000 following booster immunization.
Conclusion: This study demonstrates that EV71 VP1 mRNA-LNPs induce antigen-specific humoral immune responses in mice, providing preliminary evidence of their potential as an antigen delivery approach for EV71 and supporting further studies in vaccine development.
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Personal views expressed by the contributors in their articles are not necessarily those of the Journal of Associated Medical Sciences, Faculty of Associated Medical Sciences, Chiang Mai University.
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