Author ORCID Identifier

https://orcid.org/0000-0001-5736-0005

Semester

Summer

Date of Graduation

2023

Document Type

Dissertation

Degree Type

PhD

College

School of Medicine

Department

Not Listed

Committee Chair

Fredrick Heath Damron

Committee Co-Chair

Mariette Barbier

Committee Member

Brian Boone

Committee Member

Clinton Garret Cooper

Committee Member

Erik Hewlett

Abstract

Antimicrobial resistance (AMR) has emerged as a global crisis, posing significant challenges to the treatment and management of infectious diseases. The escalating prevalence of antibiotic-resistant bacteria has made many antimicrobial therapies ineffective, necessitating the development of alternative strategies to combat microbial infections. The rise of multidrug-resistant pathogens, such as Pseudomonas aeruginosa and Serratia marcescens, further exacerbates this problem, as these pathogens exhibit high levels of resistance to multiple antibiotics and cause severe infections in vulnerable populations. In response to the need for alternative therapeutic approaches, monoclonal antibodies (mAbs) have emerged as promising candidates for combating AMR. In this work, we aimed to harness the potential of mAbs as alternative therapeutics against P. aeruginosa and S. marcescens, while also describing the challenges associated with their development and implementation.

Chapter 2 investigates the development of mAbs against S. marcescens, which is a Gram-negative pathogen of concern, particularly among intravenous drug users. ShlA is a key virulence factor, which lyses host cells and inhibits innate immunity. Vaccination and challenge models with ShlA demonstrated protection against infection, indicating the protective potential of these antibodies. However, challenges in antigen preparation and screening methodologies hindered the generation of ShlA-specific mAbs. In Chapter 3, we successfully generated a comprehensive panel of mAbs targeting the spike protein of SARS-CoV-2. Characterization studies such as peptide-based epitope mapping provided insights into their binding profile. Overall, the work in Chapters 2 and 3 provided the foundation and framework for the development of mAbs against P. aeruginosa.

Chapter 4 focuses on the development of mAbs against P. aeruginosa, a significant nosocomial pathogen responsible for a wide range of infections. To generate mAbs against P. aeruginosa, we utilized a distinct immunization strategy by culturing P. aeruginosa with ammonium metavanadate, which induces cell envelope stress responses. This approach yielded two notable antibodies, WVDC-0357 and WVDC-0496. These antibodies demonstrated specific binding to the O-antigen lipopolysaccharide (LPS) of P. aeruginosa and exhibited bactericidal activity independent of immune effector cells. In murine models, these antibodies effectively reduced bacterial burden and mitigated the inflammatory response. This research highlights the potential of these mAbs as therapeutic interventions against P. aeruginosa and contributes to the ongoing development of effective antibody-based strategies against P. aeruginosa.

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