Abstract 1801 Characterization of Hsc70 Interactions in the Modulation of LPS/TLR4 Production of TNFα

The characterization of Heat Shock Cognate Protein 70 (Hsc70) interactions in RAW264.7 macrophages can aid in understanding its role in the Toll-Like Receptor 4 (TLR4) production of the key inflammatory cytokine Tumor Necrosis Factor α (TNFα). The TLR4 pathway is heavily regulated to maximize the efficiency of the innate immune response to bacterial lipopolysaccharide (LPS). The nature of Hsc70 as a chaperone protein encourages the study of any client interactions it undergoes within this critical pathway. Work by our lab and others has shown over 200 separate possible proteins that may interact with Hsc70 and affect TNFα secretion. In this work, RAW264.7 macrophages were cultured with LPS concentrations ranging from sub- to supra-physiological conditions. Collection of cell lysates and conditioned media at various time points allowed for validation of Hsc70-binding proteins through immunoprecipitation. These candidates can then be screened for TNFα activation to assess the respective degrees of the LPS/TLR4 inflammatory response. These results will provide insights into biochemical intricacies of the regulation of the LPS/TLR4 inflammatory response. Research reported in this publication was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103424-20.