Life Sciences Commons^™

The Response Of M0, M1, And M2 Raw246.7 Macrophage Cell Line To Hsv-1 Infection In Vitro, Amani Mohammed Alhazmi

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Herpes Simplex Virus Type 1 (HSV-1) infection occurs through the epithelial cells of the skin or mucous membranes. The beginning of the primary infection is rapid and is characterized by pain in the mouth, salivation, and submandibular lymphadenitis. The infected mucosa produces numerous, small and red lesions known as cold sores, however, many cases are asymptomatic. After the primary infection HSV-1 moves through the nerve to stay in trigeminal ganglia and to cause a recurrent infection from time to time. In the early hours of the HSV-1 infection, the cytokines produced by infected cells are critical in the stimulation of …

Hsv-1 Replication In Different Raw 264.7 And J774.1 Macrophage Phenotypes And Macrophage Viability Following Hsv-1 Infection, Yousef Nifaj Alanazi

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HSV-1 is a ubiquitous virus capable of causing lifelong latent infection. The virus contains a large double strand DNA genome covered by an icosahedral capsid. HSV-1 is a cytolytic virus which can cause a lethal infection. The virus possesses critical protein ICP0 which is capable of interfering with host cell signaling and eventually prevent cell apoptosis. Innate immunity plays a crucial role in defense against HSV-1 infection and macrophage plays a significant role in the innate immune system. Macrophage cells can alter their behavior depending upon certain stimuli and tissue environments. Naive macrophage (M0) cells can be polarized to a …

The Effects Of Socs1, Socs3 And Hsv-1 Infection On Morphology, Cell Viability And Rab7 Expression In Polarized M1 And M2 Raw 264.7 Murine Macrophages, Jessica Renee Hey

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HSV-1 causes a life-long infection in its host and has evolved multiple strategies to facilitate infection and evade the immune response. This virus has been found to enter cells by both endocytosis and fusion. The way the virus exploits endocytosis is not fully understood. Recent studies have uncovered roles of Rab GTPases, key regulators in intracellular membrane trafficking pathways, in distinct steps of the HSV-1 life cycle (Raza et al., 2018). This study will focus on analyzing the levels of the late endosomal regulator Rab7 expression in macrophages infected with HSV-1. Revealing the effect of virus on the levels of …

Virus Production And Cell Viability Of Hsv-1-Infected Murine Keratinocytes (Hel-30) Co-Cultured With Murine Macrophages (Raw 264.7), Barry Graffagna

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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 …

Effects Of Myrrh On Hsv-1 Using Plaque Assay, Badrieah Mohammad Alamri

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Herpes simplex virus type 1 (HSV-1) is a highly infective human pathogen which infects a wide range of population in North America and worldwide. HSV-1 infection has two phases, lytic and latent. Recurrence of HSV-1 is a major challenge to clinicians to control the infection especially in immune depleted individuals. Acyclovir (ACV) is an antiviral drug used to treat HSV-1 infection. Low solubility of ACV in water, mutation of viral thymidine kinase, and mutation of viral DNA polymerase are major problems that cause usage limitations of ACV. Myrrh has been used as an analgesic and anti-inflammatory natural product in middle …

Development Of A Co-Culture System To Mimic The Transfection Of Hsv-1 From Keratinocytes To Neuronal Cells, David A. Dixon

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To more closely mimic the in vivo progression of HSV-1 a novel in vitro method was created. In this study HEL-30 keratinocytes were infected with a 0.1 MOI of HSV-1 and plated unto a membrane with 8μm diameter pores. The membrane insert was then placed into culture wells that contained Neuro-2a cells (N2A). The neurites from the neuronal cells made cell-to-cell contact with the infected HEL-30 cells and the virus was transmitted into the neuronal cells. To determine if infection occurred in the N2A cells the cells were lysed and the lysate incubated with Vero cells to titrate virus plaque …

The Effect Of Hsv-1 Infection On Differentiated And Polarized U937 Cells, Allolo Dreiwish Aldreiwish

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Herpes simplex virus type 1 (HSV-1) challenges the host immune system through several mechanisms (Frey, et al., 2009). In vitro, U937 cells (human macrophage-like precursor cell line) are not susceptible to HSV-1 infection when they are not differentiating (Lopez-Guerrero and Alonso, 1997). Differentiation of these cells resistance can abrogates their resistance to HSV-1 (Tenney and Morahan, 1991). In this study, we examined the effect of HSV-1 infection on differentiated and polarized U937 cells. U937 cells are differentiated to M0 cells.

Then, M0 cells are polarized to distinct phenotypes, M1 or M2. M1 are proinflammatory macrophages while M2 are anti-inflammatory cells. …

Establishment Of A Quiescent Infection Of Hsv-1 In L929 Fibroblasts Using A Mitotic Inhibitor And Ifn-Γ, Neelam V. Shinde

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The goal for this study was to determine if a quiescent infection of HSV-1 could be induced in murine fibroblasts L929 by treating them with the anti-mitotic agent 5-fluoro 2'deoxy uridine (FUDR) alone and with interferon-γ. Since neurons are post-mitotic and exhibit a lower metabolic rate than other cells, fibroblasts were treated with FUDR to induce a post-mitotic state. The cell cycle arrest of fibroblasts would decrease the thymidylate metabolism and impair HSV-1 replication. An evaluation of cytopathic effects of FUDR was used to determine the optimal concentration which arrests cell growth and inhibits viral replication. Image J program developed …

Suppressor Of Cytokine Signaling (Socs) 1 & 3 Expression In Hsv-1- Infected And Interferon-Γ-Treated Neuro-2a Cells, Melinda Jones

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This study examined the effects of HSV-1 infection and IFN-γ treatment on Neuro-2A cells. HSV-1 induces expression of SOCS1 and SOCS3 in infected cells, inhibiting the ability of these cells to produce the pro inflammatory, antiviral cytokine IFN-γ (Nowoslawski and Benveniste, 2011). SOCS1 and SOCS3 levels were determined in IFN-γ-treated cells, virus-infected cells, and cells that were both IFN-γ-treated and virus-infected. Results were compared with untreated, uninfected control cells. Flow cytometry data analysis showed a slight decrease in SOCS1 and SOCS3 protein levels in cells treated with IFN-γ for 6 hours compared to control cells. A significant decrease in SOCS1 …

Distribution Of Cellular Interferon Beta (Ifn-Β) In Murine Fibroblast Cell Lines Upon Infection Of Hsv-1, Rachael E. Curtis

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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 …

A Potential Strategy To Maintain Hsv-1 In A Latent State: Use Of Immunoregulatory Peptide Mimetics, Nasrin Majidi

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This study reviews the role of interferon-gamma (IFN-gamma) in HSV-1 latency. CD8+ T cells inhibit the reactivation of HSV-1 in trigeminal ganglia (TG) by production of IFN-gamma. Although CD8+ T cells include all the cytotoxic apparatus for cytotoxicity, latently infected neuronal cells are not killed by CD8+ T cells. The CD94-NKG2a molecule on CD8+ T cells, binds to Qa-1b (a MHC class I like molecule) present on neuronal cell to inhibit CD8+ T cells cytotoxicity. Other studies have also determined that the IFN-gamma peptide mimetic 95-132 inhibits HSV-1 replication in the same way as IFN-gamma in infected cells. IFN-gamma mimetic …