The Immungenicity and Cross-Neutralizing Activity of Enterovirus 71 Vaccine Candidate Strains

Xia Xiao,Yebin Yu,Yuyi Liao,Haifeng Xiao,Yuan Li,Jikai Zhang,Jiankai Liu,Kangping Zhou,Meirong Gu,Junqiang Xu,Yingzhi Xu,Kun Cai,Gaimei Zhang,Guoshun Li,Peiyu Zeng

doi:10.15212/zoonoses-2021-0008

Abstract

Objective: This study aimed to evaluate enterovirus 71 (EV-A71) vaccine candidate strains, including their genotypes, immunogenicity and cross-neutralization capacity. Methods: From clinical samples, EV-A71 strains were separated by using Vero cells. Six strains were chosen for vaccine candidates, and the sequences were analyzed. To detect the immunogenicity of the strains, we used them to immunize NIH mice at 0 and 14 days. Cytopathic effects (CPE) were examined to determine the EV-A71 neutralizing antibody (NTAb) titer 14 d after the first and second inoculations. To evaluate the cross-neutralizing capacity of the EV-A71 vaccine candidate strains, we tested serum immunized mice with ten EV-A71 genotype strains. Results: Six EV-A71 vaccine candidate strains were identified, all belonging to sub-genotype C4, the prevalent genotype in China. The sequence similarity of the VP1 regions of the six candidate vaccine strains and three approved inactivated vaccines was 97.58%–97.77%, and the VP1 amino acid similarity was 98.65%–99.33%. Experiments were performed to evaluate the immunogenicity and cross-neutralizing activity of the EV-A71 vaccine candidate strains. The strains had good immunogenicity 14 d after two immunizations, inducing an NTAb titer ranging from 1:94 to 1:346. The NTAb seroconversion rates 14 d after one immunization were above 80% (except HB0007), and significantly increased immunogenicity of EV-A71 strains was observed post-inoculation. Furthermore, our candidate vaccine strains had broad cross-neutralizing activity after challenge with ten sub-genotypes of EV-A71. The highest NTAb titer/lowest NTAb titer ratios of sera against EV-A71 sub-genotypes were 8.0 (JS0002), 8.0 (JS0005), 21.3 (HB0005), 21.3 (HB0007), 10.7 (HB0040) and 8.0 (GD0002), respectively. Conclusions: Our EV-A71 strains had good immunogenicity and cross-neutralization activity, and have the potential to serve as vaccine strains for multivalent hand, foot and mouth disease vaccines.

Highlights

Enterovirus 71 (EV-A71), a single-stranded RNA virus in the Picornaviridae family, has been associated with a range of diseases, including hand, foot and mouth disease (HFMD) and central nervous system complications
We identified six EV-A71 vaccine candidate strains belonging to sub-genotype C4, which is currently prevalent in China
The results revealed that the EV-A71 strains could be divided into three genotypes: A, B and C.The EV-A71 vaccine candidate strains belonged to genotype C4 (C4a) (Fig 1), which remains predominant in mainland China.The sequence similarity of the VP1 regions among the six candidate vaccine strains and the reference strains was 97.58%–97.77%, and the VP1 amino acid similarity was 98.65%–99.33%.The amino acids at sites 146, 220, 226, 237, 241, 283, 289 and 293 differed (Table 1)

Summary

Introduction

Enterovirus 71 (EV-A71), a single-stranded RNA virus in the Picornaviridae family, has been associated with a range of diseases, including hand, foot and mouth disease (HFMD) and central nervous system complications. The EV-A71 BrCr strain (A sub-genotype) was first isolated from a woman with encephalitis in 1969 in California [5], and was the only A genotype among EV-A71 virus strains. In the 1980s, the B1 and B2 sub-genotypes became epidemics in Europe and America [6,7,8,9]. the C1 sub-genotype gradually replaced B2 in the late 1980s to early 1990s. In 1997, hundreds of children died from HFMD due to the C2, B3 and B4 sub-genotypes in the Asia-pacific region. In 2004–2005, C4 replaced the B4 sub-genotype as the main genotype in Taiwan [10]. In mainland China, C4 has remained the predominant subgenotype since its first isolation in 1997 [3,11,12]

Methods

Results

Discussion

Conclusion