Abstract
Improving vaccine immunogenicity by targeting antigens to dendritic cells has recently emerged as a new design strategy in vaccine development. In this study, the VP1 gene of foot-and-mouth disease virus (FMDV) serotype A was fused with the gene encoding human immunodeficiency virus (HIV) membrane glycoprotein gp120 or C2-V3 domain of hepatitis C virus (HCV) envelope glycoprotein E2, both of which are DC-SIGN-binding glycoproteins. After codon optimization, the VP1 protein and the two recombinant VP1-gp120 and VP1-E2 fusion proteins were expressed in Sf9 insect cells using the insect cell-baculovirus expression system. Western blotting showed that the VP1 protein and two recombinant VP1-gp120 and VP1-E2 fusion proteins were correctly expressed in the Sf9 insect cells and had good reactogenicity. Guinea pigs were then immunized with the purified proteins, and the resulting humoral and cellular immune responses were analyzed. The VP1-gp120 and VP1-E2 fusion proteins induced significantly higher specific anti-FMDV antibody levels than the VP1 protein and stronger cell-mediated immune responses. This study provides a new perspective for the development of novel FMDV subunit vaccines.
Highlights
Foot-and-mouth disease (FMD) is an acute, severe, and highly contagious disease that is caused by foot-and-mouth disease virus (FMDV), which infects cloven-hoofed animals such as cattle, pigs, and sheep
Restriction enzymes were purchased from New England Biolabs (NEB); the transfection Cellfectin5 II Reagent and Grace’s Insect Medium were purchased from Invitrogen; the mouse anti-His tag monoclonal antibody and horseradish peroxidase- (HRP-)conjugated goat anti-mouse IgG were purchased from Abbkine (California, USA)
The synthesized gene products for the VP1 proteins and the VP1-gp120 and VP1-E2 recombinant fusion proteins were digested at the BamHI and HindIII recognition sites and cloned into the pFastBac 1 vector to construct the recombinant transfer vectors pFastBac-VP1, pFastBac-VP1gp120, and pFastBac-VP1-E2, respectively (Figure 1(a))
Summary
Foot-and-mouth disease (FMD) is an acute, severe, and highly contagious disease that is caused by foot-and-mouth disease virus (FMDV), which infects cloven-hoofed animals such as cattle, pigs, and sheep. Vaccination is the most reliable and effective means of preventing and controlling FMD. Traditional FMD vaccines play an important role in the prevention and control of FMD, they present a number of shortcomings, such as incomplete inactivation of the virus and escape of live viruses from vaccine production facilities [3, 4]. The development of safe and effective new genetically engineered vaccines is required for the prevention, control, and eventual elimination of FMD in the future. The immune effects of these new genetically engineered vaccines are not superior to those of traditional inactivated vaccines. Vaccine research has focused on the adoption of new design strategies to BioMed Research International further improve the immunogenicity of these new genetically engineered vaccines
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