seed grant competition

Developing a promising vaccine candidate for porcine reproductive and respiratory syndrome
Jianrong Li, Food science and technology Porcine reproductive and respiratory syndrome (PRRS), caused by a virus (PRRSV), is considered the most economically devastating viral disease in global agriculture. It is estimated that annual losses associated with PRRS amount to half a billion dollars in the United States. Two genotypes of PRRSV, European (type 1) and North American (type 2), have been identified. Commercial PRRSV vaccines are currently available but do not effectively protect against multiple PRRSV strains. A successful vaccine has several components. First, the vaccine should be safe for pigs; second, the vaccine should trigger strong immunities; and finally, the vaccine should cross-protect against both types of PRRS. The project team used vesicular stomatitis virus (VSV) as the vector, or means, to deliver the PRRS vaccine. VSV is commonly used as a vector because it can deliver a broad range of viral genes into cells or animals. The VSV vector is particularly attractive for developing vaccines against three types of viral pathogens: viruses that cause persistent infection such as HIV; that are highly infectious such as Ebola; and that cannot be grown in cell culture such as human norovirus. Recently, VSV vectored HIV entered phase I in human clinical trials. The team aims to develop VSV as a vehicle to induce proteins that trigger defensive immune responses against PRRSV. The team went through several steps in its search for an effective PRRS vaccine. They first tried to show that live attenuated VSV would be an excellent vaccine vector for PRRS. Researchers developed a genetically engineered (recombinant) methyltransferase (MTase)-defective VSV as the vaccine vector. The VSVs messenger RNA possesses a cap structure that is essential for viral replication and gene expression. MTase is an enzyme that methylates the cap structure. Wild-type VSV caused death and weight losses in animal models, but MTasedefective VSV lost virulence in animals. These data suggest that MTase-defective VSV is a safe vaccine vector. The team then engineered and characterized an MTasedefective VSV expressing PRRSV protective antigens. To enhance the PRRSV vaccine, the team also cloned porcine heat shock gene (HSP70) into the VSV vector. HSP70 can enhance both innate and adaptive immune responses. All the engineered viruses grew slower than wild-type virus in baby hamster kidney cells. The results showed that a selection of engineered VSV expressing PRRSV antigens was successfully recovered; these viruses reproduced slowly in cell culture; and PRRSV antigens and the porcine heat shock protein were expressed by the VSV vector. Researchers also aimed to determine if VSVs expression of a single PRRSV antigen was sufficient to induce specific immune responses against PRRSV. Recombinant viruses expressing single PRRSV antigens were inoculated into animals, and the team found that they triggered only a low level of T cell response, which is not sufficient against PRRSV infection. Thus, the final step was to enhance immune responses by having VSV express multiple PRRSV antigens. The team found that PRRS-specific antibodies significantly improved when VSV expressed either two or three antigens. Ultimately, VSV expressing two antigens stimulated the strongest immune responses and is a promising vaccine candidate against PRRSV.
Jianrong Li

It is estimated that annual losses associated with PRRS amount to half a billion dollars in the United States.

www.oardc.ohio-state.edu/seeds

34

SEEDS: The OARDC Research Enhancement Competitive Grants Program