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Document Type

Thesis - University Access Only

Award Date

1992

Degree Name

Master of Science (MS)

Department

Biology and Microbiology

First Advisor

David A. Benfield

Abstract

The early interactions of porcine rotaviruses with MA-104 cells were examined in this study. The objectives were to define the optimal physical and chemical conditions for adsorption to MA-104 cells, the interaction of these viruses with sialic acid containing glycoproteins, and the mechanism of entry into MA-104 cells. Using plaque and fluorescent focus assays, the effects of time, cell age, temperature, and pH on virus adsorption were determined. Results indicated that optimal adsorption of the Ohio State University (OSU) and the Gottfried strains of porcine rotavirus to MA-104 cells was achieved by adsorption for 90 min at 37°C in media adjusted to a pH of 8 to 9. The optimal age of MA-104 cells to use for virus growth was dependent on the assay used to measure virus replication. In the plaque assay 3-day-old MA-104 cells produced the highest number of plaque forming units, whereas in the fluorescent focus assay 6- to 8-day-old cells produced the highest number of fluorescent focus units. Our studies also found that the OSU strain of rotavirus attaches to sialic acid residues on glycoproteins or glycolipids on the cell surface of MA-104 cells in a manner similar to the simian SA-11 strain of rotavirus. In contrast the Gottfried strain of rotavirus did not bind to sialic acid, but to a different binding moiety. The lysosomotropic agents (ammonium chloride and chloroquine) and the endocytotic inhibitor monodansylcadavarine (MDC) are known to inhibit the replication of viruses, such as VSV and poliovirus, that depend upon receptor-mediated endocytosis for cellular entry. The replication of OSU rotavirus was not inhibited by these compounds, indicating· that OSU does not enter MA-104 cells via receptor-mediated endocytosis. This would be consistent with previous studies indicating that the human and rhesus strains of rotaviruses enter cells via direct membrane penetration.

Library of Congress Subject Headings

Swine -- Virus diseases
Host-virus relationships

Format

application/pdf

Publisher

South Dakota State University

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