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Quantification and characterization of high-affinity membrane receptors for tumor necrosis factor on human leukemic cell lines

✍ Scribed by Peter Scheurich; Ugur Ücer; Martin Krönke; Klaus Pfizenmaier

Publisher
John Wiley and Sons
Year
1986
Tongue
French
Weight
815 KB
Volume
38
Category
Article
ISSN
0020-7136

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✦ Synopsis

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 '251-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 TNFbinding revealed the existence of high-affinity membrane receptors (Kd 1.5 x lo-" PI) and approximately 3,000 binding sitedcell on both U937 and K562, representing 2 cell lines with high and low TNF sensitivity, respectively. Disuccin-imidyl-suberate cross-linking of receptor-bound 12SI-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 TNFalpha 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.

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