Graduate Studies, UNL

 

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

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First Advisor

Jay Reddy

Degree Name

Doctor of Philosophy (Ph.D.)

Committee Members

Bin Yu, Raul Barletta, Rodrigo Cruz

Department

Integrative Biomedical Sciences

Date of this Version

3-2-2026

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 Jay Reddy

Lincoln, Nebraska, May 2026

Comments

Copyright 2026, Kiruthiga Mone. Used by permission

Abstract

Freund’s adjuvants have been extensively used in experimental research, and they modulate immune responses through various mechanisms. While both complete Freund’s adjuvant (CFA) and incomplete Freund’s adjuvant (IFA) contain the same mineral oil base, CFA contains the killed extract of Mycobacterium tuberculosis (M. tb) as an additional ingredient. Because the CFA contains mycobacterial content similar to that of Bacillus Calmette-Guérin (BCG), which induces trained immunity (TI), we tested the hypothesis that CFA may elicit TI-like features. To test this hypothesis, we investigated the metabolic and epigenetic reprogramming induced by CFA, with IFA and BCG as controls, in inbred A/J mice. Using untargeted liquid chromatography coupled with high-resolution mass spectrometry, we analyzed the serum metabolites in mice immunized wth CFA and IFA, which were distinct in each. However, by evaluating the bone marrow cells obtained from immunized mice, we observed that the CFA-induced metabolic and epigenetic changes resembled TI-like features, which were absent in the IFA group. These findings suggest that mycobacterial components in CFA contribute to durable reprogramming of innate immune cells. However, our efforts to extend these observations to outbred mouse models were limited by significant variations noted in the immune subsets. Nonetheless, based on our data, we proposed that metabolic reprogramming can be harnessed to enhance the efficacy of vaccines and tumor immunotherapies. In that direction, we made a pioneering observation that the attenuated Coxsackievirus B3 vaccine virus, termed mutant 10 (Mt10), has the potential to be used as an oncolytic virus for cancer treatment. Because BCG contains M. tb, similar to CFA, which has been used in cancer therapies, we expect that the metabolites induced by CFA and/or Mt10 virus could be potentially used as metabolic adjuvants to modulate anti-tumor responses in follow-up studies.

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