Mechanism of release of soluble forms of tumor necrosis factor/lymphotoxin receptors by phorbol myristate acetate-stimulated human THP-1 cells in vitro

The mechanism involved in the release of the soluble forms of 55 and 75 kDa TNF and lymphotoxin (LT) membrane receptors was studied in a continuous human monocytic cell line, THP-1, in vitro. THP-1 cells were found to spontaneously release soluble forms of both 55 and 75 kDa TNF/LT receptors. Release was up-regulated by PMA, and optimal release was achieved at 10(-8) M PMA. Serine protease inhibitors such as PMSF,3,4 dichloroisocoumarin, N alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK), and N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) were found to inhibit the production of both soluble TNF/LT receptors. PMSF (2 mM) also blocked receptors shedding from paraformaldehyde-fixed THP-1 cells coincubated with conditioned media from PMA-stimulated THP-1 cells. Colchicine at 1 and 10 microM stimulated the production of both soluble TNF/LT receptors, but the PMA-induced release of both soluble TNF/LT receptors was inhibited. It appears that the PMA-induced release of soluble TNF/LT receptors involves serine proteases in the extracellular space where the soluble parts of the TNF/LT receptors are cleaved directly off the cell membrane.