Generation and validation of monoclonal antibodies against hamster cytokines, chemokines and T cell activation markers 3213

Abstract Description Hamsters are excellent models of SARS CoV-2 infection and are highly susceptible to a range of human viral infections. However, the availability of reagents to track immune responses in the hamster model are extremely lacking. We developed monoclonal antibodies to better understand and characterize the immune response in hamsters, including antibodies against, cytokines, chemokines and T cell activation markers. A Luminex™ assay was developed to detect cytokines and chemokines from hamster peritoneal exudate cells (PEC) activated with toll like receptor (TLR) agonists and validation of antibodies against T cell markers was performed using flow cytometry on naïve and activated splenocytes. PEC and splenocytes secrete IL-6, TNF-a and IL-12 p40 after treatment with LPS and poly IC. Splenocytes also secrete IFN-g, IL-4, IL-2 and IL-10 after treatment with pokeweed mitogen (PWM). Further validation is ongoing, testing activated cells for expression of CXCL-10, CXCL-1, IFN-b and IL-17. Splenocytes were incubated with antibodies to hamster CD44, CD62L, CD25 and CD69. Naïve cells demonstrated both CD62L high and CD62L low populations and were CD44+, similar to expression patterns on murine splenocytes. Upon activation with PWM or Concanavalin A, splenocytes increased expression of CD25 and CD69, activation markers present at low levels on naïve cells. Together, these reagents will open new avenues of investigation for the immunology community in infection models in the hamster. Funding Sources Supported by NIH/NIAID SBIR Contract #75N93022C00044 Topic Categories Veterinary and Comparative Immunology (VET)