Abstract
The development of a negative marker vaccine against the foot-and-mouth disease virus (FMDV) will enhance the capabilities to differentiate vaccinated from infected animals and move forward in the progressive control pathway for the control of FMD. Here, we report the development of mutant FMDV of Asia1 with partial deletion of non-structural proteins 3A and 3B and characterization of their infectivity and protection response in the guinea pig model. The deleted FMDV Asia1/IND/63/1972 mutants, pAsiaΔ3A and pAsiaΔ3A3B1 were constructed from the full-length infectious clone pAsiaWT, the viable virus was rescued, and the genetic stability of the mutants was confirmed by 20 monolayer passages in BHK21 cells. The mutant Asia1 viruses showed comparable growth pattern and infectivity with that of AsiaWT in the cell culture. However, the AsiaΔ3A3B1 virus showed smaller plaque and lower virus titer with reduced infectivity in the suckling mice. In guinea pigs, the AsiaΔ3A3B1 virus failed to induce the disease, whereas the AsiaΔ3A virus induced typical secondary lesions of FMD. Vaccination with inactivated Asia1 mutant viruses induced neutralizing antibody response that was significantly lower than that of the parent virus on day 28 post-vaccination (dpv) in guinea pigs (P < 0.05). Furthermore, challenging the vaccinated guinea pigs with the homologous vaccine strain of FMDV Asia1 conferred complete protection. It is concluded that the mutant AsiaΔ3A3B1 virus has the potential to replace the wild-type virus for use as a negative marker vaccine after assessing the vaccine worth attributes in suspension cell and protective efficacy study in cattle.
Key points
• Deletion mutant viruses of FMDV Asia1, developed by PCR-mediated mutagenesis of NSP 3A and 3B1, were genetically stable.
• The growth kinetics and antigenic relatedness of the mutant viruses were comparable with that of the wild-type virus.
• Vaccination of guinea pigs with the deletion mutant viruses conferred complete protection upon challenge with the homologous virus.
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The data will be made available upon reasonable request.
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Acknowledgements
The authors thank the Director, ICAR-IVRI, Izatnagar and Joint Director, ICAR-IVRI, Bengaluru, for facilitating the research work. The authors also acknowledge SEPPIC-Air Liquide Healthcare Specialty Ingredients for providing Montanide ISA-201 VG adjuvant.
Funding
The work was supported by CAAST-ACLH (NAHEP/CAAST/2018–19) of ICAR-World Bank funded National Agricultural Higher Education Project (NAHEP). The first author was a recipient of UGC-Maulana Azad National Fellowship (Ref No.F117.1/201415/MANF201415CHRMIZ36717).
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H.L.: part of the PhD work, production of deletion clones, and characterization; S.E. and J.K.B.: cell culture and rescue of the virus. Initial infectious cDNA clone modification; R.P.T.S. and A.S.: serotyping differentiating ELISA; LPBE; N.K., G.R.R., and V.B.: statistical analysis, presentation of the graph, and TEM; P.S., M.P., and V.P.: animal experimentation and qPCR; H.J.D.: conception of the idea, overall execution, and manuscript preparation. All authors read and approved the manuscript.
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The animal experimental protocol was approved by the Institutional Animal Ethics Committee (IAEC) vide Letter F. No.8–56/Vol II/RCSS/2018–19/2 and carried out according to the guidelines of the Committee for the Purpose of Control and Supervision of Experiments in Animals (CPCSEA), Government of India.
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Lalzampuia, H., Elango, S., Biswal, J.K. et al. Infection and protection responses of deletion mutants of non-structural proteins of foot-and-mouth disease virus serotype Asia1 in guinea pigs. Appl Microbiol Biotechnol 106, 273–286 (2022). https://doi.org/10.1007/s00253-021-11692-2
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DOI: https://doi.org/10.1007/s00253-021-11692-2