Graduate Studies, UNL

 

Dissertations and Doctoral Documents, University of Nebraska-Lincoln, 2023–

Accessibility Remediation

If you are unable to use this item in its current form due to accessibility barriers, you may request remediation through our remediation request form.

First Advisor

Hiep L. X. Vu

Degree Name

Doctor of Philosophy (Ph.D.)

Committee Members

Angela Pannier, Asit Pattnaik, Dustin Loy, Sarah Sillman

Department

Integrative Biomedical Sciences

Date of this Version

2-10-2026

Document Type

Dissertation

Citation

A dissertation presented to the faculty of the Graduate College at the University of Nebraska in partial fulfillment of requirements for the degree Doctor of Philosophy

Major: Integrative Biomedical Sciences

Under the supervision of Professor Hiep L. X. Vu

Lincoln, Nebraska, December 2025

Comments

Copyright 2026, Sushmita Kumari. Used by permission

Abstract

Avian influenza virus (AIV) remains a major challenge for global poultry health, food production, and agricultural economies. Both low-pathogenic (LPAIV) and highly pathogenic (HPAIV) strains continue to circulate and evolve, creating an ongoing need for vaccines that can be updated quickly and provide broad protection. Although DNA vaccines offer a safe and flexible approach to rapidly incorporate new antigens, their use in poultry has been limited by poor delivery efficiency and the vulnerability of naked DNA to degradation. Lipid nanoparticles (LNPs), which have proven highly effective for mRNA delivery, offer a promising solution, yet their potential for DNA delivery in avian species is largely unexplored. This dissertation investigates a lipid nanoparticle-formulated DNA vaccine platform encoding hemagglutinin (HA) proteins from AIV and evaluates its immunogenicity in pre-hatch and post-hatch chickens, along with its protective efficacy using LPAIV a surrogate model. Our findings demonstrated that the LNP formulation enhanced plasmid DNA delivery in chickens, thereby markedly reducing the antigen dose required to elicit immune response. In addition to improving immunogenicity, we also showed that the LNP formulation can efficiently co-encapsulate multiple antigen-encoding plasmids and induce balanced antibody responses without the evidence of antigenic interference, supporting its potential application as a multivalent vaccine strategy. Intramuscular immunization of chickens with the LNP-DNA vaccine provided complete protection against challenge with an LAPIV strain H5N3 strain. More importantly, in-ovo inoculation of the LNP-DNA resulted in expression of the transgene in the embryos without affecting their survival, underscoring its potential for incorporation into large-scale hatchery vaccination programs. This dissertation provides evidence that LNP-DNA can be a viable approach for controlling AIV in chickens.

Download

Included in

Virology Commons

Share

COinS