PHYSIOLOGICAL CONCENTRATIONS OF TNF-ALPHA AND IL-1-BETA RELEASED FROMREPERFUSED HUMAN INTESTINE UP-REGULATE E-SELECTIN AND ICAM-1

Intestinal ischemia-reperfusion (I/R) causes local and distant tissue injury via neutrophil (PMN) activation and adhesion, Endothelial cell adhesion molecules (E-selectin, ICAM-1) mediate the adhesion and trans migration of PMN in the microcirculation. Expression of these receptor s is influenced by cytokines. To determine the physiologic concentrati ons of two specific cytokines involved in I/R, tumor necrosis factor ( TNF) and interleukin-1 (IL-1), human intestinal segments were exposed to 30 min of ischemia followed by reperfusion. Venous effluent samples were obtained; enzyme immunoassays measured maximum concentrations of TNF (30.5 +/- 1.0 pg/ml) and IL-1 (59.0 +/- 6.0 pg/ml). Cultured huma n endothelial cells were then exposed to physiologic concentrations of human recombinant TNF (10 pg/ml) and IL-1 (10 pg/ml), individually an d in combination. Flow cytometric analysis of receptor expression demo nstrated upregulation of E-selectin as early as 2 hr (P < 0.05) with m aximum effects at 4 hr. At 4 hr, E-selectin expression (% shift from b aseline) was greater with TNF and IL-1 combined (50.9 +/- 2.9, P < 0.0 1) than with either cytokine alone (TNF 34.6 +/- 4.0; IL-1 23.5 +/- 4. 0, P < 0.01). ICAM-1 receptor expression began at 4 hr with maximum ef fects at 24 hr, ICAM-1 expression after TNF and IL-1 exposure (15.4 +/ - 1.3, P < 0.001) was also greater than TNF (10.9 +/- 0.3, P < 0.01) o r IL-1 (3.1 +/- 1.5) alone, TNF and IL-1 are present in venous effluen t in concentrations capable of increasing PMN adhesion in the microcir culation. These findings support a role for these cytokines in local a nd distant organ injury from I/R. Since combined effects are greater t han either cytokine alone, antagonism of both TNF and IL-1 may be requ ired for a therapeutic benefit in clinical applications. (C) 1996 Acad emic Press, Inc.