Continued development of an immunological toolkit and profile for bottlenose dolphins
Abstract
Knowledge of the cetacean immune system as well as the diagnosis of specific etiologic agents of disease are currently lacking in cetacean medicine, specifically bottlenose dolphins. Marine mammal clinicians often rely on complete blood count and biochemistry results and the changes observed in inflammatory markers over time to determine when a dolphin is inflicted by an infectious disease agent and whether treatment is effective. Additionally, characterization of the dolphin immune system and response to infectious diseases is currently hampered by the lack of species-specific reagents needed for further immune system investigation. The objectives ofthis proposal are to address both issues in dolphin medicine. We aim to develop a predictive diagnostic model utilizing dolphin-specific cytokines, serum metabolomics, serum proteomics, the complete blood count inflammatory markers, and biochemistry inflammatory markers to better determine the etiologic agent of infectious diseases in dolphins. To train the model, serum samples from dolphins with known causes of disease (fungal infections, bacterial infections, and viral infections) will be analyzed for differences in cytokine profiles, serum metabolomics profiles, and serum proteomics profiles. Statistical analyses of the results and training of the model will provide a predictive diagnostic model that is able to provide the likely etiologic agent of disease in an ill dolphin with a certain level of confidence if successful. By improving diagnostic capabilities with less invasive methods, treatment strategiescan be tailored based on the most likely etiologic agent causing the illness and implemented more rapidly, thus lowering the risk of developing resistant microbes, improving the response to treatment and decreasing morbidity and mortality in ill dolphins. To address the lack of understanding of the cetacean immune system, we aim to continue to develop dolphin-specific cytokine primers for targeted cytokines including interleukin-1 alpha, interleukin-1 beta, interleukin-12, interleukin-23, interleukin-18, interferon-alpha,interferon-beta, chemokine (C-C morph) ligand-2, and chemokine (C-X-C morph) ligand-9; as well as develop dolphin monoclonal and polyclonal antibodies that recognize leukocyte differentiation molecules to characterize leukocyte subpopulations. The same methodologies that have been used to develop polyclonal antibodies for recombinant dolphin tumor necrosis factor-alpha, interferon-gamma, interleukin-1 receptor antagonist, interleukin-2, interleukin-4, interleukin-8, interleukin-10, interleukin-13, interleukin-17, and chemokine (C-X-C morph) ligand-10 will be used in the new reagent development for the previously listed new cytokine assays. Recombinant proteins will be expressed in yeast and/or mammalian expression systems and proteins will be isolated and utilized to immunize mice and rabbits to produce monoclonal and polyclonal antibodies of interest, respectively. Cytokine levels can be measured in order to identify modifications or alterations in the dolphin immune system as a result of environmental insults as well as serving as diagnostic tools in characterizing immune responses to pathogens and/or vaccines. If successful, dolphin-specific reagents produced as a result of this proposal will be available to other marine mammal clinicians and researchers through Kingfisher Biotech, Inc. for use in clinical medicine and research.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Nov 21, 2023
- Source ID
- N000142412000
Entities
People
- Abby M McClain
Organizations
- National Marine Mammal Foundation
- Office of Naval Research
- United States Navy