Applied Microbiology and Biotechnology

, Volume 103, Issue 8, pp 3367–3379 | Cite as

Immunogenicity and protective efficacy of recombinant proteins consisting of multiple epitopes of foot-and-mouth disease virus fused with flagellin

  • Baofeng Cui
  • Xinsheng Liu
  • Peng Zhou
  • Yuzhen Fang
  • Donghong Zhao
  • Yongguang ZhangEmail author
  • Yonglu WangEmail author
Biotechnological products and process engineering


Many recent studies have shown that flagellin fused to heterologous antigens can induce significantly enhanced humoral and cellular immune responses through its adjuvant activity. Therefore, in this study, two key B cell epitopes and a truncated VP1 (ΔVP1) protein from foot-and-mouth disease virus (FMDV) were expressed as flagellin fusion proteins in different patterns. Specifically, ΔVP1 and two duplicates of two key B cell epitopes (2×B1B2) were fused separately to the C-terminus of flagellin with a universal exogenous T cell epitope to construct FT (Flagellin-Truncated VP1) and FME (Flagellin-Multiple Epitopes). In addition, the D3 domain of flagellin was replaced by ΔVP1 in FME, yielding FTME (Flagellin-Truncated VP1-Multiple Epitopes). The immunogenicity and protective efficacy of the three fusion proteins as novel FMDV vaccine candidates were evaluated. The results showed that FT, FME, and FTME elicited significant FMDV-specific IgG responses at 10 μg/dose compared with the mock group (P < 0.05), with FTME producing the highest response. No significant differences in the antibody response to FTME were observed between different immunization routes or among adjuvants (ISA-206, poly(I·C), MPLA, and CpG-ODN) in mice. In addition, at 30 μg/dose, all three fusion proteins significantly induced neutralizing antibody production and upregulated the levels of some cytokines, including TNF-α, IFN-γ, and IL-12, in guinea pigs. Importantly, all three fusion proteins provided effective protective immunity against FMDV challenge in guinea pigs, though different protection rates were found. The results presented in this study indicate that the FTME fusion protein is a promising novel vaccine candidate for the future prevention and control of foot-and-mouth disease.


FMDV Epitope Flagellin Recombinant fusion Vaccine 



This study was supported by the National Key Research and Development Program (2016YFD0501505), the Agriculture Research System of China (CARS-35), the Central Public Interest Scientific Institution Basal Research Fund (Y2016CG23), and the National Natural Science Foundation of China (31602095).

Compliance with ethical standards

Conflict of interests

The authors declare that they have no conflict of interest.

Ethics statement

All guinea pig experiments were performed in a biosafety level 3 laboratory at the Lanzhou Veterinary Research Institute (LVRI) at the Chinese Academy of Agricultural Sciences (CAAS). This study was approved by the Institutional Animal Use and Care Committee of the CAAS. All guinea pigs used in the present study were humanely bred during the experiments and were euthanized at the end of the study.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Baofeng Cui
    • 1
    • 2
    • 3
  • Xinsheng Liu
    • 1
    • 2
  • Peng Zhou
    • 1
    • 2
  • Yuzhen Fang
    • 1
    • 2
  • Donghong Zhao
    • 1
    • 2
  • Yongguang Zhang
    • 1
    • 2
    Email author
  • Yonglu Wang
    • 1
    • 2
    Email author
  1. 1.State Key Laboratory of Veterinary Etiological Biology, OIE/National Foot and Mouth Disease Reference Laboratory, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research InstituteChinese Academy of Agricultural SciencesLanzhouChina
  2. 2.Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhouChina
  3. 3.Lanzhou Institute of Biological Products Co., Ltd. (LIBP), a subsidiary company of China National Biotec Group Company Limited (CNBG)LanzhouChina

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