Location: Foreign Animal Disease Research
Project Number: 8064-32000-060-02-T
Project Type: Trust Fund Cooperative Agreement
Start Date: Jun 1, 2013
End Date: Sep 30, 2016
This collaborative research project seeks to identify the host immune mechanism that may be responsible for protection against the infection with African Swine Fever Virus (ASFV) virulent strains. Specific objectives include: 1. Analyze the quantitative and/or qualitative differences in tissues and cell types involved in early replication of the attenuated vaccine strains compared with virulent parental virus by immunofluorescence confocal microscopy (ICM). 2. Study the profile of locally produced pro-inflammatory chemical mediators (PCM) and the activation of any other significant host genes during infection with delta 9GL virus strains compared with virulent parental ASF. 3. Assess the effect of different PCM when used as therapeutics in ASFV infected animals.
1. Infected tissues will be characterized to identify the cell types responsible for virus replication in the pre-viremic phase of infection. These cells will be stained using several cell markers. Sections will be processed with different primary and secondary antibodies and examined using confocal microscopy. 2. Comparison of transcriptional and proteomic profiles between the attenuated and virulent parental viruses may provide critical information about innate mechanisms mediating early protection after vaccination. Detection will be performed using tonsils, mandibular lymph nodes, retropharyngeal lymph node and spleen. Dection will be conducted by confocal immunofluorescence microscopy and examination of the differential gene activation patterns by real time RT-PCR. 3. The effect of different PCM when used as therapeutics in ASFV infected animals will be assessed. Host factors differentially expressed during infection with attenuated ASFV strains will be assessed as therapeutics in ASFV infections. The host factors will be cloned in effect of the different PCMs on the replication of ASFV will be first tested in vitro; using primary cell cultures of swine macrophages. The effect of PCMs will be assessed in terms of reducing the virus yields. Those PCMs showing a significant effect in vitro will be further tested in vivo.