Meeting Abstract
Immunology research often focuses on quantification of specific cells types and molecules using monoclonal or polyclonal antibodies. A major limitation in ecoimmunology and wildlife immunology is the lack of species-specific reagents for non-traditional study species. The unprecedented success of checkpoint molecule (e.g. PD-1, CTLA-4) blockade in human cancer immunotherapy in recent years suggests that understanding receptor-ligand interactions is critical to understanding immune function. This talk will demonstrate how recombinant proteins were rapidly developed to characterize the Tasmanian devil immune system and the potential immune evasion pathways used by the devil facial tumor to subvert immune defenses. Fluorescently-tagged PD-1, CD80, 4-1BB, CD200, CD200R1, CD47 recombinant proteins were used to confirm that receptor-ligand interactions observed in humans and rodents are conserved in Tasmanian devils. Fluorescently-labelled recombinant cytokines, including IFN-γ, IL-2, IL-6, IL15- IL-21, TNF-α, and VEGFA were also produced. Initial testing demonstrated that Fluorescently-labelled IFN-γ can induce upregulation of MHC-I on devil facial tumor cells on par with unlabelled IFN-γ, suggesting that the fluorescent labels do not affect protein function. Ongoing testing will determine if these fluorescently-labelled cytokines can be used to map cytokine receptor expression. Following completion of our receptor-ligand studies, the recombinant proteins can be used to produce monoclonal or polyclonal antibodies for key proteins identified in the receptor-ligand studies. Importantly, these techniques can be quickly adapted to most eukaryotic species and allow unprecedented insight into the development and regulation of the immune system of non-traditional study species.
