|Eberle, Kirsten - Iowa State University|
|Venn-watson, Stephanie - National Marine Mammal Foundation|
|Jensen, Eric - Us Navy Marine Mammal Program Biosciences Division, Space And Naval Warfare Systems Center Pacific|
Submitted to: PLoS One
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/1/2017
Publication Date: 12/22/2017
Citation: Hofstetter, A.R., Eberle, K.C., Venn-Watson, S.K., Jensen, E.D., Porter, T.J., Waters, T.E., Sacco, R.E. 2017. Monitoring bottlenose dolphin leukocyte cytokine mRNA responsiveness by qPCR. PLoS One. 12(12):e0189437. https://doi.org/10.1371/journal.pone.0189437.
DOI: https://doi.org/10.1371/journal.pone.0189437 Interpretive Summary: This study tracked longitudinal changes in the potential for bottlenose dolphin blood cells to produce immune signals after a strong non-specific stimulus. Blood was collected over the course of seven months from four dolphins and tested for thirteen different immune signals. The results cluster the thirteen signals into three groups based on magnitude. These data provide a foundation for future studies by outlining normal ranges and variability of these thirteen immune signals.
Technical Abstract: Veterinarians caring for bottlenose dolphins (Tursiops truncatus) in managed populations utilize blood-based diagnostics to monitor animal health. Blood can also be sampled from wild bottlenose dolphins for research or monitoring. Use of qPCR to assess cytokine expression patterns by peripheral blood mononuclear cells (PBMC) offers a rapid method to supplement currently available blood tests with information on immune status. Full realization of this potential requires establishment of ranges of cytokine expression levels in bottlenose dolphins. We surveyed four dolphins over the span of seven months by serial bleeds. PBMC were stimulated with phytohaemagglutinin (1, 5, and 10 µg/mL) and concanavalin A (1 µg/mL) for 24 or 48 H in vitro. RNA from these cultures were probed by qPCR using Tursiops truncatus-specific primers (IL-1a, IL-1ß, IL-1RA, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-13, IL-18, IFN-' and TNF-a). Two blood samples from an additional bottlenose dolphin diagnosed with bronchopneumonia add further perspective to the data. The results demonstrate that despite inter-animal differences, the magnitude of mitogenic response generally clusters the tested cytokines into three groups. The data provide a reference for the selection of target cytokine mRNAs and their expected range of responses in future studies.