Clin Exp Vaccine Res.  2014 Jan;3(1):100-109. 10.7774/cevr.2014.3.1.100.

Development of porcine respiratory and reproductive syndrome virus replicon vector for foot-and-mouth disease vaccine

Affiliations
  • 1College of Veterinary Medicine and Veterinary Science Research institute, Konkuk University, Seoul, Korea. virus@konkuk.ac.kr

Abstract

PURPOSE
Foot-and-mouth disease (FMD) is an economically important global animal disease. To control FMD virus (FMDV) outbreaks, a lot of different novel approaches have been attempted. In this study, we proposed a novel porcine reproductive and respiratory syndrome virus (PRRSV) as a replicon vector to express FMDV structural protein.
MATERIALS AND METHODS
PRRSV infectious clone (PRRSVK418DM) was used to develop an expression vector through the reverse genetic manipulation of PRRSV; FMDVP12A3C gene of serotype O was synthesized and used for an antigen. MARC-145 cells (African green monkey kidney epithelial cell line) were used for electroporation mediated transfection. The transfection or the expression of P12A3C and N protein of PRRSV was analyzed by either replicon containing PRRSV alone or by co-infection of helper PRRSV.
RESULTS
We constructed PRRSVK418DM replicon vector containing FMDVP12A3C, and genome sequences were confirmed by subsequent sequence analysis. In vitro expression of P12A3C and PRRSV N protein was confirmed by immunofluorescence antibody assay using antibodies specific for PRRSV N protein (anti-PRRSV N MAb), FMDV-VP1 (anti-VP1 MAb).
CONCLUSION
The results indicate that PRRSV replicon vector can be a promising novel vector system to control FMDV and useful for vaccine development in the future.

Keyword

Replicon; FMDV; PRRSV; Genetic vectors

MeSH Terms

Animals
Antibodies
Cercopithecus aethiops
Clone Cells
Coinfection
Disease Outbreaks
Electroporation
Epithelial Cells
Fluorescent Antibody Technique
Foot-and-Mouth Disease*
Genetic Vectors
Genome
Kidney
Porcine respiratory and reproductive syndrome virus*
Replicon*
Sequence Analysis
Transfection
Viruses
Antibodies
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