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Evaluating novel antibodies for cytokine profiling in the guinea pig model
Lea Maristela1, Alyssa Longworth1, David F Ackart1
1Mycobacteria Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO, United States.
Abstract:
The guinea pig model of Mycobacterium tuberculosis (Mtb) infection offers distinct advantages compared to conventional mouse models. Namely, the guinea pig model develops circumscribed granulomas with central necrosis which most closely resembles that of human granulomas. However, a limitation of this model is the lack of available reagents to characterize the Mtb immune response. To better leverage this representative model, a set of recombinant rabbit monoclonal antibodies targeting guinea pig cytokines were generated and validated. Using established stimulation conditions for primary guinea pig cells capable of producing specific cytokine profiles, the reactivity and specificity of these antibodies to their biological target were evaluated. These antibodies were then evaluated individually for direct ELISA and immunohistochemistry or part of a pair for sandwich ELISA and ELISpot techniques. Utilized in a low-dose aerosol model of Mtb in the guinea pig, the utility of these antibodies to detect specific immune cytokines generated in response to infection is demonstrated. Collectively, these antibodies will provide a more comprehensive understanding of immune kinetics, vaccine response, and comparative immunology across the guinea pig model of tuberculosis and other infectious diseases.