Date of Award

1-1-2001

Embargo Period

4-18-2024

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Microbiology and Immunology

College

College of Graduate Studies

First Advisor

Steven D. London

Second Advisor

Robert J. Boackle

Third Advisor

J. Robert Cantey

Fourth Advisor

Jean-Michel Goust

Fifth Advisor

Lucille London

Abstract

Achieving protective immunity at mucosal sites is critically important for effective immunization against pathogens that colonize the host via the wet epithelium. We utilized reovirus 1/L infection of mice to study immunization strategies for the induction of optimal humoral and cell-mediated immunity at mucosal surfaces. The humoral and cell-mediated responses at mucosal and systemic sites were monitored following upper respiratory tract, or systemic inoculation. Utilizing these models, we found that the combined upper and lower respiratory tract inoculation resulted in optimal humoral immunity proximally (in the respiratory tract), distrally (in the oral, gastrointestinal, and urogenital tracts), and systematically. This immune response induces neutralizing antibodies and could be boosted following secondary inoculation. In addition to these findings, we also demonstrated that combined upper and lower respiratory tract inoculation resulted in optimal cell-mediated responses in the gastrointestinal tract and systematically. Using a novel reovirus-specific bromodeoxyuridine incorporation assay, we found that the following combined upper and lower respiratory tract inoculation there was increased proliferation of CD4 and CD8 (helper and cytotoxic) T-lymphocytes in the spleen and draining lymph nodes. Taken together this data indicated that combined upper and lower respiratory tract inoculation induces optimal humoral and cell-mediated immunity. Finally, we monitored the proliferative responses to individual reovirus gene segments following systemic or upper and lower respiratory tract inoculation. Using this system we found that the response in systemic tissues was directed at reovirus outer capsid protein regardless of the route of inoculation. In contrast, after mucosal inoculation the response in the draining lymph nodes was directed at an inner capsid protein. Therefore these results suggest that differential antigen processing and/or presentation occurs at musical vs. systemic sites. This data has important clinical implications for vaccine delivery since induction of the upper and lower respiratory tract in human will likely require intranasal and aerosol delivery of antigen. These results also suggest that the selection of epitope(s) to be included in the vaccine must take into account the route of inoculation.

Rights

Copyright is held by the author. All rights reserved.

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