Thesis Title

Characterization of Attachment and Receptor Isolation of Infectious Pancreatic Necrosis Virus to Chse-214 Cells

Date of Graduation

Spring 1990

Degree

Master of Science in Biology

Department

Biology

Committee Chair

Christopher Field

Subject Categories

Biology

Abstract

Infectious Pancreatic Necrosis Virus (IPNV) is a bisegmented, double stranded RNA virus with a protein capsid and no envelope and is classified in the group, Birnaviridae. Although much is known about viral genes and proteins, the initial stages of attachment to susceptible cells are not known and are the focus of this research. The attachment of ³⁵S methionine labelled viruses to Chinook Salmon Embryo cells (CHSE-214) was assayed after treatment of the monolayer with various proteolytic enzymes, reducing agents and specific blocking agents. IPNV attached optimally at a pH of 7.2-7.4 within 10 to 15 seconds at 23°c. Treatment of the monolayer with chyrotrypsin and neuraminidase decreased viral attachment at all concentrations tested. Protease at 8U/mL increased viral attachment, but higher concentrations reduced attachment. Fewer viruses absorbed to Concanavalin A, lentil or fetuin treated cells, indicating the cellular receptor sites may contain a structural sialo-glycoprotein similar to glycophorin A. The IPNV receptor is suggested to reside in a coated pit covered by a masking protein, which is removed by low concentrations of protease. Preliminary research suggests the receptor protein is solubilized by mild detergent treatment and will react with labelled viruses in vitro.

Copyright

© Nora Sherry Jones

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