The expression of specific membrane receptors for TNF-alpha was determined on various human leukemic cell lines differing in their sensitivity to the growth-inhibitory activity of TNF-alpha. Binding studies with 125I-labelled TNF-alpha indicated specific binding in 8/10 cell lines with approximately 10-fold differences in the quantity of TNF-alpha bound by these distinct cell lines. Scatchard analyses of TNF-binding revealed the existence of high-affinity membrane receptors (Kd 1.5 X 10(-10) M) and approximately 3,000 binding sites/cell on both U937 and K562, representing 2 cell lines with high and low TNF sensitivity, respectively. Disuccinimidyl-suberate cross-linking of receptor-bound 125I-TNF-alpha and SDS-PAGE of membrane preparations of either U937 or K562 cells suggest a single receptor protein with an apparent molecular weight of 76 kDa. Comparison of the TNF-alpha binding capacity versus in vitro growth inhibition provides evidence that sensitivity to TNF-alpha is determined both at the level of receptor expression and at a post-receptor level. IFN-gamma strongly enhanced the TNF-alpha-mediated growth inhibition of 3 sensitive cell lines, but had no effect on 7 other leukemic cell lines with little or no TNF sensitivity. No correlation was found between this enhancement of TNF sensitivity and the IFN-gamma-mediated increase in TNF-cell membrane receptors, suggesting that IFN-gamma predominantly exerts its synergistic effect distal to TNF-binding.

A large difference in the number of gamma-IFN receptors was found on a variety of human tumor cell lines with a range of 0.4 to 15 X 10(3) binding sites/cell. The receptor number did not correlate with the potential responsiveness of the cells to gamma-IFN, in regard to either growth inhibition or induction of HLA-DR expression, two independently regulated gamma-IFN effects. However, with respect to HLA-DR expression it was found that the gamma-IFN sensitivity of inducible cell lines depended on the number of receptors, so that with increasing number of receptors present on these cells lower doses of gamma-IFN were required for induction of the gamma-IFN effect.