Virus Production and Cell Viability of HSV-1-infected Murine Keratinocytes (HEL-30) Co-cultured with Murine Macrophages (RAW 264.7)
Nancy Bigley (Advisor), Marjorie Markopoulos (Committee Member), Dawn Wooley (Committee Member)
Master of Science (MS)
Keratinocytes are the most abundant type of cell in the outer layer of skin, the epidermis, and provide barrier against pathogens from invading. However, Herpes Simplex Virus Type 1 (HSV-1) targets these keratinocytes for infection, and later infects neurons to establish lifelong latency. The keratinocytes stimulate the innate immune system to engage and to destroy the virus. Among the cells of the innate immune system to respond to the viral invasion is the macrophage. In this study, RAW 264.7 macrophage and HEL-30 keratinocyte monolayers were challenged in vitro with HSV-1 at a multiplicity of infection (MOI) of 0.1 to investigate the individual responses. A co-culture was established using a ratio of 1:5 with the macrophages and keratinocytes, respectively after the keratinocytes were challenged with HSV-1 with an MOI of 0.1. The viabilities and the viral titers of both monolayers and the co-culture were recorded after 24, 48, and 72 hours from the initial infection with HSV-1. The viabilities of both monolayers decreased over time to confirm HSV-1 infection in both cell lines. Further confirmation was made with the results of the viral titers increasing over time for both monolayers. Interestingly, the co-culture resulted in a different outcome than of the individual cultures. The infected co-culture viabilities remained high and were comparable to the uninfected co-culture over the 72 hours. The viral titer of the co-culture was significantly lower than that of the two monolayer titers at 48 and 72 hours. However, there was a significant increase in viral production from 48 to 72 hours. This suggests the RAW 264.7 macrophage in a co-culture with HEL-30 keratinocytes were able to slow the HSV-1 infection and demonstrate its protective role within the innate immune system
Department or Program
Department of Microbiology and Immunology
Year Degree Awarded
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