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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Food Safety and Enteric Pathogens Research » Research » Publications at this Location » Publication #132490
Title: BLOCKADE OF TUMOR NECROSIS FACTOR ALPHA-MEDIATED ENDOTHELIAL ACTIVATION AND APOPTOSIS WITH PORCINE TUMOR NECROSIS FACTOR RECEPTOR 1-IG FUSION PROTEIN (19TH INT CONGR OF THE TRANSPLANTATION SOCIETY, MIAMI, FL, 8/25-30/02)

Author
item COSTA, C - ALEXION PHARMACEUTICALS
item BELL, N - ALEXION PHARMACEUTICALS
item MWANGI, S - 3625-30-15
item Stabel, Thomas
item FODOR, W - ALEXION PHARMACEUTICALS

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 8/30/2002
Publication Date: 8/30/2002
Citation: COSTA, C., BELL, N.K., MWANGI, S.M., STABEL, T.J., FODOR, W.L. BLOCKADE OF TUMOR NECROSIS FACTOR ALPHA-MEDIATED ENDOTHELIAL ACTIVATION AND APOPTOSIS WITH PORCINE TUMOR NECROSIS FACTOR RECEPTOR 1-IG FUSION PROTEIN. TRANSPLANTATION. 2002. V. 74 (SUPPL.). ABSTRACT P. 353.

Interpretive Summary:

Technical Abstract: Tumor necrosis factor alpha (TNFa) plays a role in the development of delayed xenograft rejection (DXR) by promoting endothelial cell activation, apoptosis, and the recruitment of inflammatory cells. We have developed a fusion protein containing the extracellular domain of the porcine TNF-Receptor 1 (p55) and the Fc portion of porcine IgG1 (pTNFR1Ig). Incubation of porcine aortic endothelial cells (PAEC) with 20 ng/ml hTNFa for 24 h induced expression of SLAI, SLAII, VCAM-1, ICAM-1, and E-selectin. The simultaneous addition of pTNFR1Ig at 1 ug/ml to the media completely blocked this effect. Co-treatment with 1 ug/ml pTNFR1Ig for 24 h also protected from apoptosis induced by hTNFa (20 ng/ml) + cycloheximide (2.5 ug/ml). To conduct a comparative study evaluating the efficacy of pTNFR1Ig in blocking human, porcine, and murine TNFa, we incubated PAEC with 10 ng/ml of each cytokine alone or in combination with increasing concentrations of the pTNFR1Ig fusion protein (range from 10 ng/ml to 1 ug/ml). Cell surface expression of SLAI, SLAII, and E-Selectin was evaluated by flow cytometry to monitor TNFa-induced activation. Porcine and murine TNFa led to higher expression levels of these parameters than hTNFa. Concentrations of pTNFR1Ig as low as 30 ng/ml conferred some protection from pTNFa and mTNFa activity, whereas there was little effect in blocking hTNFa at this low concentration of the receptor. By contrast, pTNFR1Ig at 50 ng/ml completely counteracted the effects of hTNFa and mTNFa while there was still evidence of pTNFa activity when this concentration of pTNFR1Ig was used.