Evidence for the existence of two forms of membrane tumor necrosis factor: an integral protein and a molecule attached to its receptor.

Abstract

Plasma membranes were isolated from thioglycolate-induced peritoneal mouse macrophages and tested directly in a 51Cr-release assay against WEHI 164 tumor cells. These membranes showed anti-TNF antibody inhibitable killing of the TNF-sensitive tumor cell line, indicating that membrane-associated TNF is present on mouse macrophages. In order to elucidate whether membrane TNF is an integral protein or a molecule attached to a receptor, cells and plasma membranes were treated with low pH buffer. A partial reduction in TNF activity was observed which could be restored by incubation with exogenous TNF. In a Western blot analysis the integral membrane TNF could be identified as the 26-kDa molecule on activated mouse macrophages. These results indicate that both forms of membrane-associated TNF exist on macrophages and are responsible for cell-mediated cytotoxicity against TNF-alpha-sensitive targets.