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. 2023 Dec;12(1):2192815.
doi: 10.1080/22221751.2023.2192815.

Rational development of multicomponent mRNA vaccine candidates against mpox

Rong-Rong Zhang  1 , Zheng-Jian Wang  1 , Yi-Long Zhu  2   3 , Wei Tang  1 , Chao Zhou  1 , Suo-Qun Zhao  1 , Mei Wu  1 , Tao Ming  1 , Yong-Qiang Deng  1 , Qi Chen  1 , Ning-Yi Jin  2 , Qing Ye  1 , Xiao Li  2 , Cheng-Feng Qin  1   4
Affiliations

Rational development of multicomponent mRNA vaccine candidates against mpox

Rong-Rong Zhang et al. Emerg Microbes Infect. 2023 Dec.

Abstract

The re-emerging mpox (formerly monkeypox) virus (MPXV), a member of Orthopoxvirus genus together with variola virus (VARV) and vaccinia virus (VACV), has led to public health emergency of international concern since July 2022. Inspired by the unprecedent success of coronavirus disease 2019 (COVID-19) mRNA vaccines, the development of a safe and effective mRNA vaccine against MPXV is of high priority. Based on our established lipid nanoparticle (LNP)-encapsulated mRNA vaccine platform, we rationally constructed and prepared a panel of multicomponent MPXV vaccine candidates encoding different combinations of viral antigens including M1R, E8L, A29L, A35R, and B6R. In vitro and in vivo characterization demonstrated that two immunizations of all mRNA vaccine candidates elicit a robust antibody response as well as antigen-specific Th1-biased cellular response in mice. Importantly, the penta- and tetra-component vaccine candidates AR-MPXV5 and AR-MPXV4a showed superior capability of inducing neutralizing antibodies as well as of protecting from VACV challenge in mice. Our study provides critical insights to understand the protection mechanism of MPXV infection and direct evidence supporting further clinical development of these multicomponent mRNA vaccine candidates.

Keywords: Mpox virus; mRNA vaccine; mouse model; multicomponent; protective antigen.

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Conflict of interest statement

No potential conflict of interest was reported by the author(s).

Figures

Figure 1.
Figure 1.
Design and characterization of MPXV mRNA vaccine candidate encoding multiple antigens. (A) Construction and encapsulation of mRNA-LNPs encoding multiple proteins of MPXV. (B) MPXV protein expression in HEK293T cells. Cells were transfected with antigen-encoded mRNAs and detected at 24 h post transfection by indirect immunofluorescence staining. Scar bar, 50 μm. (C) Representative size distribution graph of A35R-LNP, B6R-LNP, M1R-LNP, A29L-LNP and E8L-LNP.
Figure 2.
Figure 2.
Multicomponent mRNA vaccine induces a robust antibody response in mice. Groups of BALB/c mice were immunized with mRNA vaccine or placebo and boosted with an equal dose 3 weeks later. Sera sample were collected at indicated times post immunization. (A) Schematic diagram of immunization and challenge experiment. (B) VACV specific IgG antibody titres were determined by ELISA. (C) Neutralizing antibody levels against VACV were determined by PRNT assay. Data are shown as mean ± SEM. Significance was analysed by two-way ANOVA with multiple comparisons tests (ns, not significant, **p < 0.01, ***p < 0.001, ****p < 0.0001).
Figure 3.
Figure 3.
Antigen-specific CD4+ and CD8+ T cell responses following immunization. BALB/c mice were immunized i.m. with two doses of multicomponent mRNA vaccine. (A–J) Flow cytometry assay for IFN-γ, IL-2, IL-4 and TNF-α in splenocytes. Spleen was harvested and stimulated with MPXV A35R (A, F), B6R (B, G), M1R (C, H), A29L (D, I) and E8L (E, J) peptide pools 7 days after two immunizations. Data are shown as mean ± SEM. Significance was analysed by two-way ANOVA with multiple comparisons tests (ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).
Figure 4.
Figure 4.
Multicomponent mRNA vaccine protects mice from VACV challenge. (A) Groups of mice immunized with mRNA vaccine or placebo were intranasally challenged with 106 PFU of VACV. Weight changes were monitored for 20 days post infection. (B–D) Viral genome copies in nasal respiratory epithelium (B), lung (C) and throat swab (D) were determined by qPCR. (E–G) VACV titres in nasal respiratory epithelium (E), lung (F) and throat swab (G) were measured using standard plaque assay in BSC-1 cells. Data are shown as mean ± SEM. Significance was analysed by one-way ANOVA or two-way ANOVA with multiple comparisons tests (ns, not significant, ns, not significant, *p < 0.05, ****p < 0.0001).
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Grants and funding

This work was supported by the National Key Research and Development Program of China [2021YFC2302400], and the National Natural Science Foundation of China [82241069]. C.-F.Q. was supported by the National Science Fund for Distinguished Young Scholars [81925025], the Innovative Research Group from the NSFC [81621005], and the Innovation Fund for Medical Sciences [2019-I2M-5-049] from the Chinese Academy of Medical Sciences.