Source: UNIVERSITY OF ILLINOIS submitted to NRP
ELICITING CROSS-PROTECTIVE IMMUNITY TO PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME [PRRS] VIRUS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1005327
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Dec 2, 2014
Project End Date
Sep 30, 2015
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF ILLINOIS
2001 S. Lincoln Ave.
URBANA,IL 61801
Performing Department
Veterinary Research & Extension
Non Technical Summary
The most recent estimate of the economic impact of a porcine reproductive and respiratory syndrome (PRRS) outbreak to the American pork industry is $650 million dollars per year. Given the high incidence of new PRRS outbreaks, the development of effective intervention strategies to prevent and/or control such events to minimize the impact of PRRS is vital. Pork producers and their veterinarians have identified the development of vaccination approaches to establish effective cross-protective immunity as a topic of high priority in order to mitigate the occurrence of herd-immunity failure to PRRS virus. Herd-immunity failure occurs when a significant portion of a population fails to acquire sufficient levels of vaccine-induced protective immunity.Two reasons that could explain poor levels of vaccine-induced cross-protective immunity arethe presence of significant antigenic differences between the vaccine and the field virus anda vaccine that, although immunogenic, fails to stimulate the proper type of immunity capable of providing cross-protective (a.k.a., heterologous) immunity.Recent advances in mucosal adjuvant technology point to the increased efficacy of a properly formulated inactivated vaccine as a promising approach toimproving immune responses, protecting immunity, and increasing the breath of coverage for viruses with high levels of antigenic variation such as influenza virus. Accordingly, it is reasonable to expect that the application of these advances in mucosal vaccine technology will lead to the development of the next generation of safe and effective inactivated mucosal PRRS vaccines that should be capable of aiding in the control of virus load in the animal and its spread in the herd. Accordingly, we will prepare a killed PRRS virus vaccine that will be optimally adjuvanted. We expect that the administration of the inactivated and adjuvanted vaccinedelivered IN will elicit strong protective immunity against the genetically divergent virus.
Animal Health Component
35%
Research Effort Categories
Basic
35%
Applied
35%
Developmental
30%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31135101090100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3510 - Swine, live animal;

Field Of Science
1090 - Immunology;
Goals / Objectives
The goal of this project is to test the hypothesis that an optimally formulated inactivated PRRS virus vaccine delivered intranasally will elicit a strong cross-protective (heterologous) immunity.
Project Methods
To ascertain the influence of the adjuvant on the magnitude and quality of the adaptive immune response to the inactivated vaccine, groups of grower pigs will be immunized twice intranasally two weeks apart with different vaccine formulations. Whole blood and serum samples will be collected at 0, 7, 14, 21 and 28 days after immunization. Measuring the level of virus neutralizing antibodies in serum will assess the level of humoral immune response of the vaccinated animals. The cellular immune response to the vaccine will be assessed by measuring the interferon gamma response to recall viral antigen of peripheral blood mononuclear cells (PBMC) isolated from heparinized blood samples. To determine level of cross-reactivity to a genetically divergent PRRS virus, in both types of assays two different viruses will be used. The "atypical PRRS abortion storm" virus isolate NADC-20, which belongs to PRRSV lineage 8, and will be used to prepare the vaccine, will serve as the source of the homologous viral antigen for the assays. The second virus will be the PRRSV isolate LTX1, which belongs to lineage 1 and is related the Canadian origin 1-22-2 RFLP viruses and will serve as the source of a genetically divergent (heterologous) viral antigen to prepare the vaccine. Based on their ORF5 sequence, these two viruses have a 88% level of homology, which represents a high level of genetic divergence and can thus be considered to a heterologous.

Progress 12/02/14 to 09/30/15

Outputs
Target Audience:The target audience consists of scientists involved in investigating the development of effective vaccines against a highly genetically diversified virus like porcine reproductive and respiratory syndrome virus (PRRSV). In addition, swine veterinarians and producers would benefit and be very interested in any development that would lead to a better understanding on how to elicit protective immunity against PRRSV. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The personnel involved in this project acquired knowledge and experience on the methods of macrophage cell culture, the methods to grow PRRSV, and themethods necessary for virus purification, concentration and downstream analysis necessary for the preparation of viral antigen suitable for vaccination studies. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? A medium scale level production of a wild-type strain of PRRSVwas prepared using ZMAC cells and then concentrated 10-fold. The amount of viral protein in the resulting product was calculated by the densitometry using protein standards in gels stained with SYPRO Orange by quantitative imaging using a FluorChem R system. The infectious virus titer was determined before and after the concentration of the virus. The concentrated virus titer reached 109 TCID50/ml and the amount of virus protein was roughly 30 mcg of viral protein. The infectivity of the virus was eliminated by chemical inactivation using binary ethylenimine (BEI). The successful inactivation of the virus was confirmed by exposing ZMAC cells to undiluted and serially diluted BEI inactivated virus preparation. The stability of the viral antigen after inactivation was confirmed by Western blot. Thus, a batch of vaccine was prepared and is ready for vaccine formulation and subsequent administration into pigs to test its immunogenicity.

Publications