Performing Department
(N/A)
Non Technical Summary
African swine fever (ASF) is a highly contagious viral disease with mortality up to 100% in domestic swine herds. ASF outbreaks in Eurasia have resulted in tremendous economic losses to those affected countries and the introduction of ASF into US swine industry would lead to approximately $50 billion in losses. Although live attenuated vaccines have achieved partial success, safety concern and viral persistence are the major issues. In this project, rational design and evaluation of a ASF mRNA vaccine will be conducted. The goal of this study is to evaluate the engineered vaccine antigens using LNP-mRNA formulations for stimulation of strong antibody and T cell immune responses.This work will ultimately lead to development of a mRNA vaccine against ASFin aid of disease control and swine health.
Animal Health Component
100%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
(N/A)
Goals / Objectives
The goal of this project is to evaluate the immunogenicity of novel-designed vaccine antigens using mRNA technology for optimal induction of both B cell and T cell immunities against ASFV.This will be accomplished through two objectives: 1) Rational design, proteinengineering, and validation of candidate antigens in miceusing mRNA technology; 2) In vivo immunogenicity evaluation of candidate ASF mRNA vaccine in pigs.
Project Methods
Engineered vaccine candidate antigens will be formulated into LNP-mRNA and validated for antigen expression using in vitro testing methods, including flow cytometry, Western blot, confocal microscopy. LNP-mRNA will be evaluated in mice first before testing in pigs. The goal of the mouse study is to select the optimal engineering strategies and combinations for induction of strong B cell and T cell immunities. Immunogenicity assessment will be initially conducted in mice.During termination, spleen will be harvested for isolation of splenocytes.Based on the outcomes of mouse study, the best-performed candidate antigens will be administrated to 4-week-old piglets at a dosage of 30 µg/antigen/pig intramuscularly. Serum and blood samples will be collected weekly for assessing humoral immunity by ELISA. At week 5 during termination of the study, Spleen and draining lymph node will be collected for assessing cellular immunity by ELISpot and flowcytometry. Part of the tissues will be embedded in cryomold containing Tissue-Tek OCT compound for germinal center analysis by tissue imaging.