Publication Date
2011
Document Type
Thesis
Committee Members
Nancy Bigley (Advisor), Cheryl Conley (Committee Member), Barbara Hull (Committee Member)
Degree Name
Master of Science (MS)
Abstract
The hypothesis for this study was: IFN-β expression differs between murine fibroblasts (A2R1 and L929) of different mouse strains upon infection of HSV-1. Fluorescent microscopy was used to examine localization of IFN-β in the different murine fibroblast and keratinocyte cell lines after 6 hours of infection with herpes simplex virus type-1 (HSV-1). Because keratinocytes cell lines HEL-30 and PAM-212 grew in clusters, staining patterns in individual cells could not be determined. A notable difference in localization of IFN-β immune staining was seen between the two cell lines when infected with HSV-1. The A2R1 cell line showed perinuclear (PN) localization, while the L929 cell line showed nuclear localization suggesting a distinction in response to HSV by these fibroblast lines. These responses were compared with those stimulated by an inducer of Type I interferons, polyriboinosinic:polyribocytidylic acid (poly I:C). In Poly I:C-treated fibroblasts, cytoplasmic staining was observed at 6 hours after treatment.
Image J Fire program provided by National Institute of Health (NIH) was used to determine fluorescent intensities in both the keratinocyte cell lines and the fibroblast cell lines. The fluorescent intensity was significantly greater in the L929 fibroblast cell line when treated with poly IC than the untreated L929 fibroblast. There was no difference in fluorescent intensities in any of the other cell lines following poly IC treatment.
I attempted to measure IFN-β in the culture supernatants from the infected cells at different time points: 0, 6, 12 and 24 hours, from all cell lines (A2R1, L929, Hel-30 and Pam-212) using a Mouse Interferon Beta ELISA kit (range 15.6-1000pg/ml), but none was detected, except for an insignificant amount in the L929 cells stimulated with Poly IC at 24 hours. The control standards that were supplied with the kit gave the expected results.
Page Count
65
Department or Program
Microbiology and Immunology
Year Degree Awarded
2011
Copyright
Copyright 2011, all rights reserved. This open access ETD is published by Wright State University and OhioLINK.
