Author ORCID Identifier

0000-0002-0199-8699

Date of Award

Summer 8-11-2020

Degree Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biology

First Advisor

Julia Hilliard

Second Advisor

Richard Dix

Third Advisor

John Houghton

Abstract

B virus (BV, Macacine alphaherpesvirus 1), a neurotropic Simplexvirus, is enzootic in macaque monkeys, infects all of the 17 species of the macaque monkeys including Macaca mulatta, M. fascicularis, and M. nemestrina, the most extensively used nonhuman primate biomedical research models, but lifelong, mostly latent, zoonotic infection in humans often leads to severe neurologic impairment or fatal encephalomyelitis in up to 80% of reported cases. Early detection is key as no virus-eliminating therapeutics are available currently. Antivirals are available as a prophylactic measure, or as an effective therapy only if administered very early in acute, symptomatic infection. Because virus shedding is rare, serological detection of BV-specific antibodies is often the only diagnostic option for identifying BV infection. Current BV diagnostic methods are labor-intensive complex assays and even these must be performed frequently to differentiate cross-reactive BV and HSV antibodies. Hence, it is critical to identify markers of specific immune responses and virus neutralizing targets. To define the relevance of BV gB in controlling the BV infection, by accurate diagnosis and informing rational vaccine design, the hypothesis “B virus neutralization and specific antibody induction are

each dependent on unique glycoprotein B epitopes” was tested.

Epitope mapping of the neutralizing mAb identified novel mimotopes, corresponding to aa residues 115-120 and aa residues 515-525 of BVgB, that are potential targets for virus neutralization and may serve as epitope-focused recombinant protein-based BV vaccine candidates. Epitope mapping of the non-neutralizing mAb, using phage display and peptide array, revealed a B virus specific epitope that can identify anti-BV polyclonal antibodies from both BV infected humans and macaques without cross reacting with HSV1 and 2 polyclonal antibodies. Through this study I propose a novel, simple, rapid and cost-effective method of fine mapping linear epitopes using phage display and compound mutagenesis that circumvents the use of expensive peptide array and laborious mutagenesis scanning techniques. The impact of this study will inform vaccine designs that will block virus entry, as well as the development of novel strategies for early and unequivocal identification of B virus zoonotic infections in individuals with highly cross-reactive antibodies.

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