Crouse, Bethany2023-11-302023-11-302022-08https://hdl.handle.net/11299/258913University of Minnesota Ph.D. dissertation. August 2022. Major: Pharmacology. Advisor: Marco Pravetoni. 1 computer file (PDF); viii, 246 pages.Opioid use disorders (OUD) and overdose are public health crises that are worsening despite the availability of approved pharmacotherapies. Active immunization with anti-opioid conjugate vaccines is a novel therapeutic strategy to treat OUD and prevent overdose. To date, clinical studies suggest that efficacy of anti-drug conjugate vaccines is limited to a subset of individuals who can produce optimal antibody responses. To increase positive treatment outcomes and clinical success, this research program investigated several complementary strategies to increase OUD vaccine efficacy. First, mechanisms of optimal anti-opioid vaccine response are investigated by elucidating the immunological mechanisms behind a previously established interleukin-4 (IL-4) mediated increase in vaccine efficacy. These studies found that depletion of IL-4 resulted in a Type I IL-4R mediated increase in germinal center formation and germinal center T cell response which leads to increased opioid-specific antibody secreting cells, and that vaccine efficacy is dependent on a balanced Th1/Th2 T cell response in mice. Next, these results provided a blueprint for next generation anti-fentanyl vaccine formulations incorporating novel adjuvants targeting toll-like receptors (TLRs). These data show that a TLR7/8 agonist adjuvant increases vaccine efficacy in rodent and porcine models of fentanyl misuse and overdose. Third, vaccine design and immunization paradigms were assessed to optimize the efficacy of a novel carfentanil vaccine alone and in combination with a lead fentanyl vaccine. Longer linker lengths and a co-administered bivalent immunization strategy were associated with increased vaccine efficacy. Then, environmental factors contributing to the immune response are investigated by testing whether changes in the gastrointestinal microbiome would affect vaccine efficacy and whether these specific changes in the microbiome could be utilized as biomarkers. These studies revealed that changes in the microbiome in specific pathogen free or immune-experienced rodents did not affect efficacy of anti-oxycodone or anti-fentanyl vaccines. Finally, exploratory studies were performed to identify putative biomarkers that may be predictive of anti-opioid vaccine response in preclinical and clinical investigations. These studies indicate that pre-immunization concentration of IL-4 is correlated with vaccine efficacy in genetically diverse mice, and that specific cytokines may be of interest as indicators of immune response in human patients. Overall, the findings outlined in this research program support the use of novel adjuvants and predictive biomarkers to increase clinical efficacy of vaccines to treat OUD and overdose.enAdaptive ImmunityBiomarkersIL-4Opioid use disorderTLRVaccinesThesis or Dissertation