Investigating distinct immunization routes in zebrafish on mucosal immunity (VET2P.1133)

• Published in: The Journal of immunology (1950) Vol. 194; no. 1_Supplement; pp. 147 - 147.5
• Main Authors: Criscitiello, Michael, Young, Matthew, Troy, Savannah, Cory, Hope, McCauley, Naomi, Al-Kahtani, Sibba, Morgan, Thomas, Scholtz, Sheridan, Li, Ellen, Ali, Hasanain, Bains, Sarab, Briles, Brenna, Chang, Jonathan, Cruz-Vespa, Diego, Diaz Navarro, Jean, Eberle, Brady, Gonzalez, Allison, Marvin, Sophia, Nelson, Mary, Roubion, Courtney, Watts, Jiang, Chen, Pat, Basler, Julie, Minckler, Zachary, Wu, James
• Format: Journal Article
• Language: English
• Published: 01.05.2015
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.194.Supp.147.5

Abstract Model organisms such as Danio rerio are used to explore physiological reactions to stress and disease. Immune response is measured by quantifying lymphocyte presence and the production of mRNA encoding different immunoglobulin proteins. Since the method through which D. rerio is exposed to an antigen affects its immune response and the degree to which immunity is developed, we will compare the immune response after visceral or mucosal introduction of dinitrophenol hapten conjugated to Keyhole Limpet hemocyanin (DNP-KLH) in D. rerio, for which there is no published data. We will either combine DNP-KLH with Freund’s incomplete adjuvant and inject the mixture into the fish (100 uL at 1 mg/mL) or combine DNP-KLH with cholera toxin and introduce the mixture to their water (35 mg/L). Tissue samples will be collected from the immunized fish and two control groups of fish at weekly points throughout the experiment and will be analyzed for lymphocyte versus granulocyte ratios and expression of IgM, IgZ-1, and IgZ-2 levels via qPCR. We expect an increase in lymphocyte percentages for both immunized groups compared to the control groups, with a higher lymphocyte ratio in the mucosal group than the visceral group. Furthermore, we expect more mucosal IgZ isotypes in the mucosal group and higher levels of the IgM, IgZ-1, and IgZ-2 antibodies in the visceral group than in the mucosal group. For future experiments, we intend to test broader data sets, including exposure to multiple antigens.