Source: CAMBRIDGE TECHNOLOGIES LLC submitted to
UNIVERSAL SWINE INFLUENZA VACCINE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1016006
Grant No.
2018-33610-28256
Cumulative Award Amt.
$100,000.00
Proposal No.
2018-00816
Multistate No.
(N/A)
Project Start Date
Aug 1, 2018
Project End Date
Mar 31, 2019
Grant Year
2018
Program Code
[8.3]- Animal Production & Protection
Project Director
Hause, B.
Recipient Organization
CAMBRIDGE TECHNOLOGIES LLC
1525 BIOSCIENCE DRIVE
WORTHINGTON,MN 56187
Performing Department
(N/A)
Non Technical Summary
Influenza virus causes significant economic losses to the swine industry due to disease and decreased growth rates. Additionally, swine influenza viruses are zoonotic and represent a threat to public health. Both inactivated and live swine influenza virus vaccines are commercially available but are generally considered to lack efficacy to genetically diverse viruses circulating in the field. Here we will develop an influenza virus vaccine using a fundamentally different approach compared to currently licensed products. We anticipate that this vaccine will be efficacious to genetically diverse challenge viruses and stimulate effective, broad protection in growing pigs.
Animal Health Component
25%
Research Effort Categories
Basic
25%
Applied
25%
Developmental
50%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31135991101100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3599 - Swine, general/other;

Field Of Science
1101 - Virology;
Goals / Objectives
The main goals of this project are to further develop and optimize a universal swine influenza virus vaccine to protect pigs against influenza A virus. The proof of concept vaccine used a baculovirus platform to express recombinant swine influenza neuraminidase (NA) subtype 1 which was effiacious against a heterologous H1N1 challenge in pigs. This project will further develop this concept to enable commercialization. Specifically, a second neuraminidase (subtype 2) will be added to the vaccine as pigs are infected with both NA1 and NA2 containing viruses. This will broaden the vaccine protection to cover all circulating strains. In addition, the N1 and N2 proteins will be mutated such that they gain the ability to bind to sialic acids on cellular receptors. These mutations occur naturally in some strains of influenza virus and allow the virus to bind cellular receptors via NA as opposed to the typical hemagglutinin protein. We hypothesize these mutationswill enhance antigen uptake by respiratory epithelial cells. Finally, we will demonstrate vaccine efficacy using a swine vaccination/challenge model. Our proof of concept vaccine was delivered intramuscularly. Here, we plan to deliver the vaccine by the intranasal route using a mucosal adjuvant in conjunction with the mutated NA proteins. We expect that intranasal delivery of the vaccine will stimulate both mucosal and humoral immunity as compared to our intramuscular vaccine which principally stimulates humoral immunity. Following intranasal vaccination, pigs will be challenged with either a H1N1 or H3N2 virus. Protection will be assessed by reduction in clinical disease, viral shedding and histopathology. Demonstration of protection to both H1N1 and H3N2 viruses will lead to vaccine commercialization.
Project Methods
1. Mutant forms of neurmainidase 1 (NA1) and 2 (NA2) will be synthesized and cloned into a baculovirus expression platform. Protein expression will be verified.2. Mutant NA1 and NA2 will be characterized in vitro and their ability to engage cellular receptors confirmed.3. Experimental NA1 and NA2-contained vaccine will be formulated with an intranasal adjuvant and used to vaccinate pigs. Protection will be assessed with H1N1 and H3N2 challenges.

Progress 08/01/18 to 03/31/19

Outputs
Target Audience:The goal of this project was to develop a universal swine influenza virus vaccine which will be licensed with the USDA for use in swine. The mid-term target audience is regulators at the Center for Veterinary Biologics which will grant a commercial license to this product. The final target audience is the swine industry, specifically swine producers and veterinarians which raise and care for swine. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Technical staff (research scientists) at Cambridge Technologies participated in all aspects of the strain construction and characterization and testing in pigs. Likewise, faculty and student collaborators at South Dakota State University played key roles in strain construction and characterizaiton and testing in pigs How have the results been disseminated to communities of interest?These results are being held confidential as the resulting technology developed in being commercialized by Cambridge Technologies. Once licensed by the USDA Center for Veterinary Biologics the results will be widely publicized and also reported in the scientific literature What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Vaccination has been the most effective means of controlling influenza virus infections of pigs. However, protective antibody responses are largely strain-specific and are not effective against different influenza viruses. Therefore the development of a vaccine candidate that elicits broad-spectrum protection against diverse swine influenza A viruses (IAV), ie. a universal swine influenza vaccine, is critically needed. Cambridge Technologies has developed a novel recombinant swine influenza neuraminidase (NA) subunit vaccine. A recently completed proof of concept vaccine trial containing the baculovirus-expressed neuraminidase type 1 (BacNA1) protein demonstrated that our vaccine candidate delivered by intramuscular (IM) inoculation provided effective heterologous homosubtypic protection against SIV in pigs. Here we developed a second baculovirus strain that expressed NA2, BacNA2, to broaden coverage to both N1 and N2-containing influenza viruses which represent essentially all contemporary circulating IAVs. To further refine these vaccine antigens for mucosal administration via intranasal (IN) delivery, we generated mutants BacNA1 G147R and BacNA2 D151G. These mutations were previously shown to confer hemagglutinin (HA)-like receptor (sialic acid-containing cell surface proteins) binding properties to NA. Both wild type and mutant NA protein expression were verified by Western blotting and NA activity assay. They were shown to form virus like particles by electron microscopy when expressed by baculovirus. Protection afforded by IM administration of BacNA1/BacNA2 and IN administration of BacNA1 G147R/BacNA2 D151G were assessed in pigs including non-vaccinated controls as well as pigs vaccinated with conventional whole inactivated BacNA1/BacNA2 parental virus vaccines (i.e., swine H1N1 and H1N2). BacNA1/BacNA2 administered IM stimulated a robust anti-NA antibody response as seen by the neuraminidase inhibition assay while both IM and IN vaccination routes stimulated mucosal anti-influenza IgA. In contrast to non-vaccinated control and whole inactivated virus vaccines, pigs vaccinated with NA subunit vaccines were protected against heterologous H1N1 and H3N2 IAV challenges as demonstrated with reduction in IAV shedding in nasal swabs, reduction in IAV titer in bronchoalveolar lavage fluids and lung tissue, and near complete protection from lung lesions. These results, coupled with numerous completed internal trials, demonstrate broad-based immunity to IAV conferred by BacNA1/BacNA2. ?

Publications