Progress 02/15/16 to 02/14/21
Outputs
Target Audience:Colleagues who are interested in developing PRRSV vaccine, stakeholders in swine industry, government and academic. Changes/Problems:There were no major changes that affected the terms and conditions of the award. The pandemic was a disruption and my collaborator moved from Kansas to University of Illinois but these issues were dealt with and the project completed. What opportunities for training and professional development has the project provided?
Nothing Reported
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?
The main objective of this proposal is to test proof-of-principle that PIV5 is an excellent vaccine vector for swine by developing a porcine reproductive and respiratory syndrome virus (PRRSV) vaccine using PIV5 as a vector. There are three supportive objectives: Objective 1: Generating recombinant PIV5 expressing PRRSV antigens as a potential vaccine for PRRSV. In our recent work, we have generated recombinant PIV5 expressing antigens of influenza virus, respiratory syncytial virus (RSV) and rabies virus. A single dose inoculation of the recombinant viruses protects mice from challenge of respective viruses. PRRSV is an important pathogen for swine. A safe and efficacious PRRSV is needed. Based on our successful use of PIV5 as a vector for vaccine development, we hypothesize that PIV5 expressing antigens of PRRSV (PIV5-PRRSV) will be an excellent vaccine candidate for PRRSV. In this objective, we will generate PIV5 expressing antigens of PRRSV and analyze the viruses in cultured cells. Final Outcome: We have generated 11 constructs of PIV5 expressing PRRSV proteins as proposed. We have sequenced all the constructs and examined expression of PRRSV proteins in PIV5-PRRSV-infected cells. All insertions of PRRSV gene in PIV5 genome were successful except the insertion of NSP2 gene of PRRSV. We have had to modify the NSP2 gene to abolish its function to obtain a recombinant PIV5 that can express peptides of NSP2. Objective 2: Testing efficacy of PIV5-PRRSV in pigs. We will test efficacy of PIV5 in pigs. We will use different routes of administration (intranasal, IN, and intramuscular, IM). We will use single dose as well as prime-boost (homologous and heterologous) in case a single dose immunization is not sufficient. We will use commercial inactivated PRRSV vaccine as a control to compare efficacy of our vaccine candidates. We will determine critical antigens for protection if PIV5-PRRSV is effective. Final Outcome: We have performed tests of efficacy of PIV5-PRRSV vaccines in piglets as proposed. We have found that the PIV5-PRRSV vaccines generated better cellular immune responses than the commercial modified live attenuated vaccine (MLV). However, our vaccine did not reduce virus titer as the MLV vaccine. However, very excitingly, our vaccine candidates protected piglets from weight loss after virus challenge. Thus, we have succeeded in generating a PRRSV vaccine candidate that maintain the performance of swine after immunization. Objective 3: Improving efficacy of PIV5-PRRSV in pigs. Our recent studies show that modifications to PIV5 can enhance immune responses to antigens. We hypothesize that regulating host innate immune responses can lead to a better vaccine. We will further modify the vector to achieve better efficacy. Final Outcome: We have constructed additional PIV5-based PRRSV vaccine candidates that express combination of PRRSV antigens in backbones that are more efficacious in generating immune responses. Most excitingly, our new constructs were able to generate neutralizing antibody against PRRSV. In a challenge experiment, our new vaccine candidates protected against lung lesions and weight loss.
Publications
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Progress 02/15/18 to 02/14/19
Outputs
Target Audience:Colleagues who are interested in developing PRRSV vaccine, stakeholders in swine industry, government and academic. Changes/Problems:We did not make as much as progress due to contamination issues of our vaccine stocks with BVDV1. We have since cleaned the vaccines. What opportunities for training and professional development has the project provided?We recruited two graduate students, Caray Kirby and Elizabeth Wrobel. We have graduated a DVM resident with a PH.D. (Mathew Abraham). 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?We will challenge immunized pigs with a more virulent strain.
Impacts
What was accomplished under these goals?
We have made modified NSP2 antigens. We have also made more constructs targeting NSP2.
Publications
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Progress 02/15/17 to 02/14/18
Outputs
Target Audience:Colleagues who are interested in developing PRRSV vaccine, stakeholders in swine industry, government and academia. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?We have trained a Ph.D. candidate Kelsey Briggs under this proposal. In addition, we have trained undergraduate researchers Cara Kirby and Darren Diaz. Both undergraduate students are joining Ph.D. program to pursue scientific research as a career. How have the results been disseminated to communities of interest?Attended and presented poster at the USDA meeting. What do you plan to do during the next reporting period to accomplish the goals?We will challenge immunized pigs with a more virulent strain.
Impacts
What was accomplished under these goals?
Objective 1: Progress: We have generated 11 constructs of PIV5 expressing PRRSV proteins as proposed. We have sequenced all the constructs and examined expression of PRRSV proteins in PIV5-PRRSV-infected cells. All insertions of PRRSV gene in PIV5 genome were successful except the insertion of NSP2 gene of PRRSV. We have had to modify the NSP2 gene to abolish its function to obtain a recombinant PIV5 that can express peptides of NSP2. Objective 2: Progress: We have performed a test of efficacy of PIV5-PRRSV vaccines in piglets as proposed. We have found that the PIV5-PRRSV vaccines generated better cellular immune responses than the commercial modified live attenuated vaccine (MLV). However, our vaccine did not reduce virus titer as the MLV vaccine. Unfortunately, none of the vaccines reduced gross pathological effects of PRRSV infection. We suspect that the challenge virus we used was not robust enough. In next round of testing, we will use a more virulent strain of PRRSV. Objective 3: We have made modified NSP2 antigens.
Publications
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Progress 02/15/16 to 02/14/17
Outputs
Target Audience:Colleagues who are interested in developing PRRSV vaccine, stakeholders in swine industry, government and academia. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?We have trained post-doctoral fellows and graduate students in molecular virology techniques, and provided opportunities for them to attend scientific conferences. Dr. Huiling Wei is a post-doctoral fellow who has been working on the project. Her work and training under this proposal will enhance her opportunity to obtain a faculty position in veterinary school. How have the results been disseminated to communities of interest?We have presented our work at the annual USDA PI meeting in Chicago.We have presented this work in our annual USDA PD meeting and animal health worker meeting. What do you plan to do during the next reporting period to accomplish the goals?We will test efficacies of our vaccine candidates as proposed in specific aim 2 of the proposal. Objective 1: Generating recombinant PIV5 expressing PRRSV antigens as a potential vaccine for PRRSV.In our recent work, we have generated recombinant PIV5 expressing antigens of influenza virus, respiratory syncytial virus (RSV) and rabies virus. A single dose inoculation of the recombinant viruses protects mice from challenge of respective viruses. PRRSV is an important pathogen for swine. A safe and efficacious PRRSV is needed. Based on our successful use of PIV5 as a vector for vaccine development,we hypothesize that PIV5 expressing antigens of PRRSV (PIV5-PRRSV) will be an excellent vaccine candidate for PRRSV.In this objective, we will generate PIV5 expressing antigens of PRRSV and analyze the viruses in cultured cells. None next year Objective 2: Testing efficacy of PIV5-PRRSV in pigs.We will test efficacy of PIV5 in pigs. We will use different routes of administration (intranasal, IN, and intramuscular, IM). We will use single dose as well as prime-boost (homologous and heterologous) in case a single dose immunization is not sufficient. We will use commercial PRRSV vaccine as a control to compare efficacy of our vaccine candidates. We will determine critical antigens for protection if PIV5-PRRSV is effective. We will test efficacy of our recombinant PIV5 expressing PRRSV antigens in pigs next year. Objective 3: Improving efficacy of PIV5-PRRSV in pigs.Our recent studies show that modifications to PIV5 can enhance immune responses to antigens.We hypothesize that regulating host innate immune responses can lead to a better vaccine.We will further modify the vector to achieve better efficacy. We will generate additional recombinant PIV5 with improved antigens as well as modified PIV5 vectors next year.
Impacts
What was accomplished under these goals?
Porcine reproductive and respiratory syndrome, caused by porcine reproductive and respiratory syndrome virus (PRRSV), has caused tremendous economic losses to the swine industry. Hallmark symptoms of PRRS are mild to severe respiratory disease in infected newborn and growing swine, and reproductive failure in pregnant sows. At present, there is no effective vaccine for PRRSV. In this proposal, we are developing an effective vaccine for PRRSV. Objective 1: Generating recombinant PIV5 expressing PRRSV antigens as a potential vaccine for PRRSV. In our recent work, we have generated recombinant PIV5 expressing antigens of influenza virus, respiratory syncytial virus (RSV) and rabies virus. A single dose inoculation of the recombinant viruses protects mice from challenge of respective viruses. PRRSV is an important pathogen for swine. A safe and efficacious PRRSV is needed. Based on our successful use of PIV5 as a vector for vaccine development, we hypothesize that PIV5 expressing antigens of PRRSV (PIV5-PRRSV) will be an excellent vaccine candidate for PRRSV. In this objective, we will generate PIV5 expressing antigens of PRRSV and analyze the viruses in cultured cells. We have generated 11 recombinant PIV5 containing different parts of entire PRRSV genome. We have analyzed these recombinant viruses in tissue culture cells. Objective 2: Testing efficacy of PIV5-PRRSV in pigs. We will test efficacy of PIV5 in pigs. We will use different routes of administration (intranasal, IN, and intramuscular, IM). We will use single dose as well as prime-boost (homologous and heterologous) in case a single dose immunization is not sufficient. We will use commercial PRRSV vaccine as a control to compare efficacy of our vaccine candidates. We will determine critical antigens for protection if PIV5-PRRSV is effective. Not yet started. Objective 3: Improving efficacy of PIV5-PRRSV in pigs. Our recent studies show that modifications to PIV5 can enhance immune responses to antigens. We hypothesize that regulating host innate immune responses can lead to a better vaccine. We will further modify the vector to achieve better efficacy. Not yet started. OVERALL IMPACT: We have accomplished the objectives set forth in our proposal. These recombinant viruses are not only potential PRRSV candidates, they can also be used for expressing various genes of PRRSV for purposes other than vaccine development.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Poster presentation in annual USDA PI meeting in Chicago December 4, 2016.
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