Progress 03/15/16 to 03/14/19
Outputs
Target Audience:Researchers, veterinarians, herd owners and disease control responsible agents. Changes/Problems:-We attempted several times to overexpress IBV-S protein (Whole S protein) in E. coli with little success. Basically, the protein yield was low to use for polyanhydride encapsulation. To overcome the yield problem, we used the whole-inactivated IBV virus instead of E. coli purified protein. The obtained results confirmed that the project goals were accomplished despite this change in choice of IBV antigens. What opportunities for training and professional development has the project provided?In this project, a graduate student, a scientist and research assistant are working on various aspects of paratuberculosis vaccine. I work with the team members to accomplish the project goals and at the same time develop their technical, professional and presentation skills. I supported participation of team members in both local and national meetings where they presented their research findings. When experiment conclude, I will work with the students to present the project findings through scientific reports and presentations in national and international conferences. In general, training is provided in different aspects of virology and genetic manipulations (Basic bacteriology, genetics and genomics). In addition, more chances are given for training on different approaches to analyze host immune responses (Immunology). How have the results been disseminated to communities of interest?Two reports submitted. Presentation in the following meetings. Next Generation Polyanhydride Nanovaccine Platform Technology For Poultry. GRC on Nanoscale Science and Engineering for Agriculture and Food Systems. June 3-8, 2018. South Hadley, MA. Nanovaccines in Sickness and in Health. Illinois Institute of Technology Seminar. October 16, 2018. Nanovaccines for animal diseases, the Polyanhydride platform technology. European Congress on Vaccine and Vaccination 2018. October 26-27, 2018. Budapest, Hungary. Nanovaccines and Nano-adjuvants for Efficient Vaccination Programs. World Vaccine Congress., April 14-17, 2019. Washington D.C. A Novel Nanovaccine With Broad Spectrum Immunity Against Different Avian Influenza Subtypes. Annual Egyptian Veterinary Poultry Association. April 20-23. Porto Marina, Alexandria, Egypt. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
-To test for the safety of PAN (Aim I), specific pathogen-free (SPF) eggs were inoculated with PAN particles loaded with Fluorescein isothiocyanate (FITC) and hatchability rate and embryo development compared with mock (PBS-inoculated) group. Normal embryo development was observed in PAN-FITC inoculated eggs and hatchability rates of PAN-FITC inoculated eggs was identical to mock inoculated eggs (100%). Furthermore, the dosage of PAN particles (10 mg) administered in this pilot experiment was 10 times more than the normal PAN-adjuvanted vaccine dose (1 mg). This pilot study demonstrates the safety associated with administration of PANs in chicken embryos. In subsequent experiments, we prepared PAN-based vaccines and inoculated 1 day old chicks with 2 mg of PAN-Flu vaccine with no untoward effect on any of the immunized chicks. To develop PAN as novel vaccine platform technology, we estimated the immune responses (Aim II) generated from 1 day old chicks immunized with PAN-S encapsulating S1 protein. In addition, we used whole inactivated IBV virus as a rapid protocol to develop an effective vaccine against IBV. The Narasimhan lab synthesized PAN-S and PAN-IBV inactivated vaccine candidates following diacid synthesis, 20:80 CPTEG:CPH copolymer using a water-oil-oil double emulsion process. Quality control (QC) for all particles were performed on each batch consisted scanning electron microscopy (SEM) indicated the size PAN-IBV size range of 80-120 nm. In addition, the release kinetics of PAN-S indicated a 20% burst of payload after 1 day of PAN-S suspension in water while the continuous release of antigen continued up to the end of observation period at 5 days post suspension in water. For estimating protective immunity of PAN vaccine candidates (Aim III), 1 day old chicks were immunized with 2 mg PAN loaded with 20 ug IBV whole virus or S1 protein via intramuscular (IM) injection. At 3 weeks post immunization, all chicks were challenged with a virulent virus strain (Ark-DPI, 6.5E9 genome copy no/bird). At 8 days post challenge, PAN-S vaccine and MLV chicks have increased serum IgY titers relative to unvaccinated chicks while PAN-IBV (whole-inactivated virus) did not (data not shown). In addition, significant high levels of viral shedding in mock (Negative control) vaccinated chicks were detected in tracheal swabs compared to those birds vaccinated with modified live vaccine MLV (Positive control) or PAN-S (only S1 protein encapsulated PAN). Clinically, animals vaccinated with PAN-S1 didn't show the full spectrum of disease progress (coughing and sneezing) compared to un-vaccinated chicks (data not shown). These results demonstrate that PAN adjuvanted S1 vaccines can elicit protective immunity in chickens when challenged with virulent strains of IBV.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2019
Citation:
1. Thukral, A., Ross, K, Hansen, C., Phanse, Y., Narasimhan, B., Steinberg, H., Talaat, A.M.* (2018). A Single Dose Polyanhydride-based Nanovaccine Against Johnes Disease. (submitted npj.Vaccines).
2. Kingstad-Bakke, B.A.; Chandrasekar, S.S.; Phanse, Y.; Ross, K.A.; Marulasiddappa, S.; Kawaoka, Y.; Osorio, J.E.; Narasimhan, B.; Talaat, A.M. Effective Mosaic-based Nanovaccines against Avian Influenza in Poultry. (Submitted, Vaccines).
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