Date of Award

2024

Document Type

Thesis

Degree Name

Bachelor of Science

Department

Biochemistry & Molecular Biol.

First Advisor

Dr. Eugene Wu

Second Advisor

Dr. Jonathan Dattelbaum

Abstract

Adenoviruses are important gene delivery vectors and causative agents for a variety of human diseases such as the common cold and gastrointestinal infections. Human adenovirus type 64 (Ad64; formerly 19c) and type 37 are associated with epidemic keratoconjunctivitis. Based upon its high homology and similar disease tropism to Ad37, we hypothesized that Ad64 would have the same protein receptor, CD46 (membrane cofactor protein), as Ad37. We show that a recombinant Ad64 containing an enhanced Green Fluorescent Protein transgene (Ad64.eGFP) enters Chinese hamster ovary cells expressing human CD46 (CHO-CD46) on the surface. Entry into human cervical carcinoma (HeLa) cells is increased by the presence of calcium, but that increase can be blocked by an anti-CD46 antibody. Ad64.eGFP gene delivery into human conjunctival epithelial (HCjE) cells can also be blocked by soluble CD46, supporting our hypothesis that Ad64 uses CD46 as a protein receptor on the surface of target cells. CD46 expression on CHO-CD46, HeLa, and HCjE cells was confirmed by flow cytometry and/or western blotting. The identification of CD46 as a receptor for Ad64 makes Ad64 an attractive candidate for gene delivery into eye cells as well as other cells that express CD46.

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