Location: Foreign Animal Disease Research
Project Number: 3022-32000-063-009-I
Project Type: Interagency Reimbursable Agreement
Start Date: Aug 14, 2019
End Date: Sep 30, 2022
Objective:
This research project seeks to develop a safe and effective commercial African Swine Fever (ASF) vaccine. The objective of this project is to apply systems biology approach by combining immunological, genomic and computational methods to investigate the mechanisms of protection and to identify protective antigens.
Specific aims are to determine the roles of antigen-specific immune mechanisms including humoral immunity via (1) virus neutralization, (2) antibody-directed cell cytotoxicity and (3) macrophage activation and cell-mediated immunity through T lymphocyte cytotoxicity in inhibiting virus infection and/or replication and to identify candidates of protective antigens. Specifically, it is proposed to test the effects of sera and immune cells from vaccinated pigs on protecting macrophages which are the main target cells of the infection in-vivo, against ASFV infection.
Approach:
1. The roles of antigen-specific immune mechanisms, including humoral immunity, will be determined through in-vitro and ex-vivo assays evaluation. The level of protection against viral infection in swine macrophages undergoing different immunological treatments will be assessed through peptide microarrays and ELISA's, optimized to detect antigen-specific antibodies. A methodology will be developed for major histocompatabilty complex (mhc) class I and II genotyping for mhc matching. T cell measurement and sorting will then be conducted by flow cytometry. The resulting identified antibodies will be tested to determine level of protection against in-vitro ASF virus infection in ex-vivo macrophages.
2. To identify protective antigens and fine-tune protective immune mechanisms that are involved, a study will be conducted comparing protected and non-protected pigs after vaccination. The antigen-specific antibody response will be measured with peptide microarray analysis and ELISA. The induction of antigen-specific t lymphocyte subsets will be assessed using flow cytometry. The differences in protection against ASF virus infection in ex-vivo macrophages between protected and non-protected pigs will then be identified.
