Document Type

Thesis

Degree Name

Master of Science (MSc)

Department

Biology

Program Name/Specialization

Integrative Biology

Faculty/School

Faculty of Science

First Advisor

Dr. Stephanie DeWitte-Orr

Advisor Role

Supervisor

Abstract

Viral double-stranded RNA (dsRNA) molecules are a potent pathogen-associated molecular pattern and play a crucial role in the innate immune response. During a viral infection, extracellular and intracellular dsRNA can initiate pathways resulting in the production of type I interferons (IFNs) and interferon-stimulated genes (ISGs). The accumulation of ISGs within a cell results in a protective antiviral state. This study used both commercially available dsRNA (poly I:C) and in vitro transcribed dsRNA molecules, based on the viral hemorrhagic septicemia virus (VHSV) genome sequence, as stimuli to investigate the effects of these molecules on the innate immune response in rainbow trout cells. The goals of the present project were to elucidate the i) IFN ii) ISG and iii) antiviral responses of fish cells to both types of dsRNA molecules. Different lengths of poly I:C and in vitro transcribed dsRNA were used to determine potential length effects of dsRNA in fish cells. The aims of the project were achieved using a functional interferon assay, an ISG-promoter reporter system, an antiviral assay, and RT-PCR. It was found that extracellular dsRNA, either poly I:C or in vitro transcribed dsRNA, is able to induce innate antiviral responses in the fish cell line, RTG-2. Consistent with mammalian studies there was a greater magnitude of immune response when cells were stimulated with longer dsRNA molecules, demonstrating dsRNA length effects in fish cells.

Convocation Year

2014

Convocation Season

Fall

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