Abstract
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. It produces severe economic losses in the livestock industry. Currently available vaccines are based on inactivated FMD virus (FMDV). The use of empty capsids as a subunit vaccine has been reported to be a promising candidate because it avoids the use of virus in the vaccine production and conserves the conformational epitopes of the virus. In this report, we explored transient gene expression (TGE) in serum-free suspension-growing mammalian cells for the production of FMDV recombinant empty capsids as a subunit vaccine. The recombinant proteins produced, assembled into empty capsids and induced protective immune response against viral challenge in mice. Furthermore, they were recognized by anti-FMDV bovine sera. By using this technology, we were able to achieve expression levels that are compatible with the development of a vaccine. Thus, TGE of mammalian cells is an easy to perform, scalable and cost-effective technology for the production of a recombinant subunit vaccine against FMDV.
Highlights
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals such as cattle, pigs, sheep and deer
Suspension growing 293-6E cells were transfected with pTT5-P12A3C or with the combination of pTT5-VP0, pTT5-VP3 and pTT5-VP1
Western blot assay of the cell lysates showed that when the pTT5-P12A3C plasmid was used for transfection, the P12A capsid precursor was successfully cleaved by protease 3C into the structural proteins VP0 (37 kDa), VP3 (23 kDa) and VP1 (23 kDa) (Figure 2)
Summary
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals such as cattle, pigs, sheep and deer. The disease is endemic in many parts of the developing world and continues to pose a serious threat to livestock industries. It is of economic importance because the presence of the disease in developing countries results in severe restrictions to international trade and an outbreak in FMD-free countries can cause billionaire losses [1]. Vaccination is still a major strategy in developing countries to control FMD [2]. Foot-and-mouth disease virus (FMDV), a member of the family Picornaviridae, genus Aphthovirus, is a non-enveloped single positive-sense stranded RNA virus and the etiological agent of the disease [3]. An update of the antigenic composition of the vaccine is required when new field strains appear
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