Source: UNIV OF CONNECTICUT submitted to NRP
PLUG-AND-PLAY MULTI-PATHOGEN VACCINE PLATFORM FOR ANIMAL DISEASE COMPLEXES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1016781
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Dec 7, 2018
Project End Date
Sep 30, 2021
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIV OF CONNECTICUT
438 WHITNEY RD EXTENSION UNIT 1133
STORRS,CT 06269
Performing Department
Pathobiology & Veterinary Science
Non Technical Summary
Animal disease complexes such as chicken respiratory disease pose a challenge for the development of vaccines due to multi-pathogen etiologies and the need to frequently update the vaccine, as in the case of influenza.We recently developed a novel plug-and-play vaccine technology that allows the rapid and efficient generation of recombinant vaccines based on vaccinia virus, at one genetic locus.We now plan to expand the platform to two genetic loci for the rapid development of multi-antigen, multi-pathogen vaccines that can be rapidly updated as needed.We will develop a multi-pathogen vaccine for respiratory disease complex in chickens expressing protective antigens for infectious bronchitis, infectious bursal disease, and avian influenza. In addition, the vaccine will have a safety feature recently developed in our laboratory for vaccinators and animal handlers. There would be much to gain from a plug-and-play platform that allows rapid generation, testing, and updating of safe multi-pathogen vaccine candidates.
Animal Health Component
34%
Research Effort Categories
Basic
33%
Applied
34%
Developmental
33%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31132991101100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3299 - Poultry, general/other;

Field Of Science
1101 - Virology;
Goals / Objectives
Animal disease complexes such as chicken respiratory disease pose a challenge for the development of vaccines due to multi-pathogen etiologies and the need to frequently update the vaccine, as in the case of influenza.Vaccinia virus (VACV) is an excellent live recombinant vaccine vector due to its thermal stability, low cost of production, and ability to elicit strong humoral and cell-mediated immune responses.We recently developed a novel plug-and-play technology that allows the rapid and efficient generation of recombinant VACVs at one genetic locus.We now plan to expand the platform to two genetic loci for the rapid development of a safe multi-pathogen vaccine for respiratory disease complex in chickens expressing protective antigens for infectious bronchitis, infectious bursal disease, and avian influenza.
Project Methods
Our plug-and-play platform will be expanded to two genetic loci for the rapid development of multi-antigen, multi-pathogen vaccines that can be rapidly updated as needed, as in the case of influenza vaccines.A new transfer vector targeting the second locus will be generated and cells will be infected with the first-locus VACV and transfected with the transfer vector to generate a new recombinant virus that will serve as the standard plug-and-play parental virus that will allow the quick sequential generation of recombinant VACVs expressing antigens at two loci.The growth kinetics of this VACV will be tested in cell culture to ensure it replicates at wildtype levels.We will then generate a multi-pathogen vaccine for respiratory disease complex in chickens expressing protective antigens for three major respiratory pathogens: infectious bronchitis virus (IBV), infectious bursal disease virus (IBDV), and avian influenza virus (AIV).A new transfer vector will be generated for expression of the S gene of IBV and the VP2 gene of IBDV at the first loci and the resulting virus will be used for the generation of the final VACV also expressing AI HA at the second loci.The resulting replication-repressible virus will replicate constitutively in the absence of antibiotics, but not in the presence of doxycycline.Finally, we will characterize the multi-pathogen vaccine candidate (expression of S, VP2, and HA proteins will be confirmed by western blot).

Progress 12/07/18 to 09/30/21

Outputs
Target Audience:Vaccine researchers, virologists, and immunologists in academia and industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided training for 2 graduate students. They have received training on molecular biology and molecular virology techniques (eg, gene design, synthesis, and cloning, transfections, infections, virus purification, and recombinant virus analyses), in addition to general virology training. How have the results been disseminated to communities of interest?Our results have not yet been disseminated widely because the idea has commercial value and the potential to be patented. In fact, the results will likely be linked to our non-provisional patent application already submitted for the EPPIC platform (Jasperse, B., C. M. O'Connell, Y. Wang, and P. H. Verardi. A method for the rapid generation of recombinant poxviruses. Non-provisional US patent application 17/365,746, filed July 1, 2021). We plan to publish results soon after the continuation-in-part (CIP) patent application is submitted. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? For objective 1, we generated parental viruses by a traditional "non-spontaneous" selection process that, as expected, takes a few months as it typically requires multiple (at least 10) serial plaque purifications to completely eliminate the original parental virus. The new strategy, dubbed EPPIC and funded by an earlier NIFA/SAES award, was published earlier in 2020 (Jasperse, B., C. M. O'Connell, Y. Wang, and P. H. Verardi. 2020. EPPIC (Efficient Purification by Parental Inducer Constraint) platform for rapid generation of recombinant vaccinia viruses. Mol. Ther. Methods Clin. Dev. 17:731-738. PMCID: PMC7177189). For objective 2, we tested the strategy by quickly generating a multi-pathogen vaccine for respiratory disease complex in chickens by the "plug-and-play" strategy and characterized its genetic stability.

Publications


    Progress 10/01/19 to 09/30/20

    Outputs
    Target Audience:Vaccine researchers, virologists, and immunologists in academia and industry. Changes/Problems:Project studies were delayed due to COVID-19 and the University policy to stop on campus research March - June, 2020. A project change was requested to extend the project period one year to 9/30/2021. What opportunities for training and professional development has the project provided?This project has provided training for 2 graduate students.They have received training on molecular biology and molecular virology techniques (eg, gene design, synthesis, and cloning, transfections, infections, virus purification, and recombinant virus analyses), in addition to general virology training. How have the results been disseminated to communities of interest?Our results have not yet been disseminated widely because the idea has commercial value and the potential to be patented (it will likely be included in a provisional patent application already submitted for the EPPIC platform).It is our goal, however, to publish results soon after. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we plan to finalize the characterization and test the genetic stability of the multi-pathogen vaccine for respiratory disease complex.

    Impacts
    What was accomplished under these goals? Work on Objective 1 (develop a plug-and-play platform at two genetic loci that allows the rapid development of candidate vaccines expressing multiple genes/antigens) was completed.We generated parental viruses by a traditional "non-spontaneous" selection process that, as expected, takes a few months as it typically requires multiple (at least 10) serial plaque purifications to completely eliminate the original parental virus.The new strategy, dubbed EPPIC and funded by an earlier NIFA/SAES award, was published earlier in 2020 (Jasperse, B., C. M. O'Connell, Y. Wang, and P. H. Verardi.2020.) EPPIC (Efficient Purification by Parental Inducer Constraint) platform for rapid generation of recombinant vaccinia viruses.Mol. Ther. Methods Clin. Dev.17:731-738.PMCID: PMC7177189.We focused our attention in Aim 2, where we tested the strategy by quickly generated multi-pathogen vaccine for respiratory disease complex in chickens by the "plug-and-play" strategy.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Jasperse, B., C. M. O'Connell, Y. Wang, and P. H. Verardi. 2020. EPPIC (Efficient Purification by Parental Inducer Constraint) platform for rapid generation of recombinant vaccinia viruses. Mol. Ther. Methods Clin. Dev. 17:731-738. PMCID: PMC7177189).


    Progress 12/07/18 to 09/30/19

    Outputs
    Target Audience:Vaccine researchers, virologists, and immunologists in academia and industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project provided training for 2 graduate students. They have received training on molecular biology and molecular virology techniques (eg, gene design, synthesis, and cloning, transfections, infections, virus purification, and recombinant virus analyses), in addition to general virology training. How have the results been disseminated to communities of interest?Our results have not yet been disseminated widely because the idea has commercial value and the potential to be patented.It is our goal, however, to publish results in the upcoming reporting periods. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we plan to finalize the parental virus, test its genetic stability, and pursue Aim 2, where we will quickly generate the multi-pathogen vaccine for respiratory disease complex in chickens by the "plug-and-play" strategy.

    Impacts
    What was accomplished under these goals? Work on Objective 1 (develop a plug-and-play platform at two genetic loci that allows the rapid development of candidate vaccines expressing multiple genes/antigens) is ongoing.We are now at the stage of generating the final parental virus by a traditional "non-spontaneous" selection process that, as expected, takes a few months as it typically requires multiple (at least 10) serial plaque purifications to completely eliminate the original parental virus.We will then test the genetic stability of the new construct and pursue Aim 2, where we will quickly generate the multi-pathogen vaccine for respiratory disease complex in chickens by the "plug-and-play" strategy.

    Publications