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Virus Production and Cell Viability of HSV-1-infected Murine Keratinocytes (HEL-30) Co-cultured with Murine Macrophages (RAW 264.7)

Graffagna, Barry
http://rave.ohiolink.edu/etdc/view?acc_num=wright1542212790178886

Abstract Details

2018, Master of Science (MS), Wright State University, Microbiology and Immunology.
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.
Nancy Bigley, Ph.D. (Advisor)
Marjorie Markopoulos, Ph.D. (Committee Member)
Dawn Wooley, Ph.D. (Committee Member)
52 p.
Immunology; Microbiology
HEL-30; HEL-30 keratinocyte; RAW 264,7; RAW macrophage; keratinocyte; macrophage; co-culture; co culture; coculture; HSV-1; HSV1; herpes simplex type 1; cell viability; viral titer; plaque assay

Recommended Citations

Citations

  • Graffagna, B. (2018). Virus Production and Cell Viability of HSV-1-infected Murine Keratinocytes (HEL-30) Co-cultured with Murine Macrophages (RAW 264.7) [Master's thesis, Wright State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=wright1542212790178886

    APA Style (7th edition)

  • Graffagna, Barry. Virus Production and Cell Viability of HSV-1-infected Murine Keratinocytes (HEL-30) Co-cultured with Murine Macrophages (RAW 264.7). 2018. Wright State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=wright1542212790178886.

    MLA Style (8th edition)

  • Graffagna, Barry. "Virus Production and Cell Viability of HSV-1-infected Murine Keratinocytes (HEL-30) Co-cultured with Murine Macrophages (RAW 264.7)." Master's thesis, Wright State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=wright1542212790178886

    Chicago Manual of Style (17th edition)

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© 2018, all rights reserved.
This open access ETD is published by Wright State University and OhioLINK.
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