Thesis
USE OF RNA-SEQ TO IDENTIFY GENES ENCODING CYTOKINES AND CHEMOKINES ACTIVATED FOLLOWING UPTAKE AND PROCESSING A CANDIDATE PEPTIDE VACCINE DEVELOPED AGAINST MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS
Washington State University
Master of Science (MS), Washington State University
01/2022
DOI:
https://doi.org/10.7273/000004498
Handle:
https://hdl.handle.net/2376/123317
Abstract
Analysis of the primary and recall responses to a 35 kD major membrane protein (MMP), encoded by MAP2121c demonstrated tri-directional signaling between the antigen presenting cell (APC), CD4 and CD8 is essential for eliciting a CD8 cytotoxic T cell (CTL) response against Mycobacterium avium subsp. paratuberculosis. As reported here, RNA-sequencing was used to initiate characterization of the signaling pathways involved in eliciting development of CD8 CTL, starting with characterization of the activation status of genes in monocyte-derived macrophages (MoMΦ) following uptake and processing MMP for presentation of antigenic epitopes to CD4 and CD8 T cells. Activation status was compared with uptake and processing of LPS, a nonspecific stimulator of macrophages. 1609 genes were identified that were upregulated and 1277 were downregulated three hours after uptake and processing MMP. No significant difference was observed in the cytokine genes selected for analysis of the signaling that must occur between APC, CD4, and CD8 for development of CTL. The initial observations indicate screening of the transcriptome should include genes involved in signaling between APC and CD4 and CD8 regardless of their activation status. Four of the genes of interest in this study, IL12A, IL12B, IL15, and IL23A, were not significantly different than control values. The initial studies also indicate MoMΦ can be included with dendritic cells and monocyte-derived dendritic cells for further analysis of the tri-directional signaling required for development of CTL.
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Details
- Title
- USE OF RNA-SEQ TO IDENTIFY GENES ENCODING CYTOKINES AND CHEMOKINES ACTIVATED FOLLOWING UPTAKE AND PROCESSING A CANDIDATE PEPTIDE VACCINE DEVELOPED AGAINST MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS
- Creators
- Michelle DeCourcey
- Contributors
- Cleverson D Souza (Advisor)William C Davis (Committee Member)Janean Fidel (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Veterinary Medicine, College of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University
- Number of pages
- 28
- Identifiers
- 99900882237101842
- Language
- English
- Resource Type
- Thesis