Project Number: 5030-32000-117-74-I
Project Type: Interagency Reimbursable Agreement
Start Date: Oct 1, 2019
End Date: Sep 30, 2021
We propose to continue the development of immune assays for Atlantic bottlenose dolphins. This will require expression of specific recombinant cytokines to be used in producing antibodies for each cytokine and for use as standards in enzyme-linked immunosorbent assay (ELISA) development. Furthermore, we propose to develop antibodies to cell-surface molecules expressed on immune cells. We will then utilize these reagents to compare cytokine responses of dolphins during respiratory infections which are similar to those seen in cattle with bovine respiratory disease complex.
We will continue to test antibodies raised against human, bovine and porcine immune molecules for cross-reactivity with Atlantic bottlenose dolphin tissue samples. This has proven to be a low-risk method of identifying antibodies that could be used in immunoassays. We will continue to develop new primers sets for measuring messenger RNA levels of dolphin cytokines. New immune reagent development will continue. In our previous agreement, we developed specific cytokine ELISA reagents. As in previous research proposals, we will express recombinant cytokines in a yeast expression system and then generate polyclonal and monoclonal antibodies to these cytokines. In this proposal, we will screen and/or produce monoclonal antibodies to specific cell surface molecules. The ultimate goal will be to produce and characterize monoclonal and polyclonal antibodies for use in several bottlenose dolphin-specific immune assays, including enzyme-linked immunosorbent assays, immunohistochemical assays and flow cytometry.