Publication Date
2016
Document Type
Thesis
Committee Members
Nancy J. Bigley (Advisor), Barbara E. Hull (Committee Member), Dawn P. Wooley (Committee Member)
Degree Name
Master of Science (MS)
Abstract
Mubarak Huraysan Almutairi. M.S. Department of Microbiology and Immunology, Wright State University, 2016. The Impact of HSV-1 Infection, SOCS1 peptide, and SOCS3 peptide mimetic on Cell Viability, Morphology, and Cytoskeleton Proteins of Unpolarized and Cytokine-Polarized M1 RAW 264.7 Murine Macrophages. The immune response against HSV-1 involves macrophages in both innate and adaptive immunity by limiting HSV-1 replication. In this study, the effects of HSV-1 challenge on cell viability, morphology, and cytoskeletal filament in unpolarized and cytokine-polarized murine RAW 264.7 macrophages at 24 and 48 hours were monitored. Because the distribution of cytoskeleton throughout the cell is critical in cell viability and function, effects of HSV-1 challenge on the organization of F-actin and microtubule (tubulin) in unpolarized and cytokine-polarized murine RAW 264.7 macrophages were monitored at 24 and 48 hours post infection. F-actin and tubulin organization were assessed by quantifying the fluorescent intensity of immunofluorescent images using ImageJ analysis. M1 polarized cells displayed a significant decrease (p, 0.001) in cell viability when compared to control cells. At 24 and 48 hours post polarization, M1 cells showed flattened, irregular shapes with intracellular vacuoles, whereas unpolarized control cells (M0) appeared rounded. Following HSV-1 infection, both M0 and M1 macrophages exhibited a rounded shape. HSV-1 infection enhanced the organization of F-actin expression by unpolarized M0 and polarized M1 cells at 24 hours post infection; decreases in F-actin expression for all cells were observed at 48 hours post infection. The effects of treatments with peptide mimetics of suppressor of cytokine signaling (SOCS) proteins, namely SOCS1 and SOCS3, on cell viability and the organization of F-actin and tubulin of uninfected and HSV1-infected unpolarized and cytokine-polarized murine RAW 264.7 macrophages were evaluated at 24 and 48 hours. Treatment with a SOCS3 peptide mimetic increased cell viability of polarized M1 and HSV-1-infected M1 RAW 264.7 cells, whereas treatments with a SOCS1 peptide mimetic decreased iv viability of these cells. These observations suggest that SOCS3 peptide mimetic functions as an anti-inflammatory (anti-apoptotic) molecule by preventing cell death. Treatment of HSV-1-infected polarized M1 cells with either SOCS1 or SOCS3 peptide mimetic increased tubulin expression (p<0.001), suggesting that mechanism increased microtubule expression, such as increasing microtubule stability as a consequence of RhoA GTPases activation by SOCS1 and SOCS3 proteins.
Page Count
62
Department or Program
Microbiology and Immunology
Year Degree Awarded
2016
Copyright
Copyright 2016, all rights reserved. My ETD will be available under the "Fair Use" terms of copyright law.
