Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 11/9/1999
Publication Date: N/A
Technical Abstract: Tumor necrosis factor receptor 1 (TNFR1) mediates cytotoxic and proliferative effects of TNF-alpha and is involved in induction of TNF-alpha induced tissue damage during disease. A soluble TNF receptor (sTNFR1) is formed by proteolytic cleavage at the cell membrane surface and circulates in blood as a truncated, soluble form of the receptor with full TNF binding capacity. This anti-TNF-alpha capacity suggests that recombinant sTNFR1 may have a potential therapeutic role in swine infectious diseases in which excess TNF-alpha causes deleterious effects. It is unknown whether salmonellosis in swine results in formation of the soluble, circulating form of TNF-alpha receptors. A panel of 5 monoclonal antibodies was produced by immunizing mice with recombinant porcine sTNFR1. Positive hybridoma antibodies were selected by ELISA based on their reactivity to recombinant porcine sTNFR1 or intact PK-15 cells. Three of the five monoclonal antibodies were of the IgG1 isotype and reacted with recombinant porcine sTNFR1. The other two monoclonal antibodies were of the IgM isotype and reacted with an unknown PK-15 cell surface antigen. The IgG1 antibodies were also shown to react with an independently cloned and sequenced porcine recombinant sTNFR1 construct expressed in E. coli and P. pastoris. Antibodies are being used to develop an ELISA to measure sTNFR1 concentrations in swine sera. We speculate that serum values of sTNFR1 may serve as a diagnostic aid in porcine systemic and gastrointestinal tract diseases, as they do in similar human disease states.