Access Type

Open Access Thesis

Date of Award

January 2022

Degree Type

Thesis

Degree Name

M.S.

Department

Immunology and Microbiology

First Advisor

Philip E. Pellett

Abstract

Human cytomegalovirus (HCMV) causes severe disease in immunocompromised individuals and is a leading cause of congenital disease. Efficient assembly of virions (viral particles) is an intricate process that requires modulation and modification of host systems. HCMV induces an extensive rearrangement of the cellular endocytic recycling compartment into the site of virion maturation and egress: the cytoplasmic virion assembly compartment (cVAC). The HCMV cVAC is a distinguishing characteristic of infection, although studies have only been performed in traditional cell culture with limited observations in infectedhuman tissues. To study the cVAC, viral and cellular markers that are discrete to their respective compartments are required. This work uses immunofluorescent microscopy to determine colocalization of two cellular and viral markers. We also propose a method using cell spheroids as a tissue-like 3D model to examine the cVAC. This work validates the concurrent use of viral IE2, a nuclear viral marker, along with the cellular Golgi-marker GM130 using the same secondary antibody in fluorescence microscopy and establishes a workflow for culture and processing of HCMV-infected spheroids.

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