MUTUAL ANTAGONISM BETWEEN INTERFERON-GAMMA AND TUMOR-NECROSIS-FACTOR-ALPHA ON FIBROBLAST-LIKE SYNOVIOCYTES - PARADOXICAL INDUCTION OF IFN-GAMMA AND TNF-ALPHA RECEPTOR EXPRESSION

We recently described mutual antagonism between IFN-gamma and TNF-alph a on human fibroblast-like synoviocytes (FLS). TNF-alpha inhibits IFN- gamma-induced HLA-DR expression and IFN-gamma blocks TNF-alpha-depende nt synoviocyte proliferation, collagenase production, and GM-CSF secre tion. To study the mechanism of antagonism we have analyzed the effect these factors on the expression of cytokine surface receptors. I-125- Labeled cytokine binding was measured on cultured FLS and the results were analyzed by Scatchard plots. Unstimulated synoviocytes expressed 9300 +/- 1560 IFN-gamma binding sites per cell. A single class of high -affinity receptor was observed (K(d) = 4.5 +/- 2.5 x 10(-10) M). TNF- alpha did not competitively inhibit I-125-IFN-gamma binding. When FLS were incubated with TNF-alpha (100 ng/ml), there was a paradoxical 49. 5 +/- 5.6% increase in the number of binding sites for IFN-gamma (P = 0.001), with no change in the K(d). Unstimulated FLS also expressed 28 50 +/- 700 TNF-alpha receptors per cells, with a single K(d) consisten t with the lower-affinity TNF-alpha receptor (7.4 +/- 0.2 x 10(-10) M) . IFN-gamma did not directly interfere with TNF-alpha binding. Preincu bation of FLS with 100 U/ml of IFN-gamma resulted in a 28.9 +/- 9.0% i ncrease in TNF-alpha receptor expression (P < 0.008), with no change i n the K(d). Low levels of the soluble 55-kD TNF receptor were detected in FLS supernatants. IFN-gamma did not effect soluble TNF receptor pr oduction. These data are the first demonstration of IFN-gamma and TNF- alpha receptors on FLS and show that TNF-alpha and IFN-gamma increase the expression of each other's receptor. Therefore the mutual antagoni sm between these two cytokines must occur through a postreceptor mecha nism.