Recombinant human tumour necrosis factor increases cytosolic free calcium in murine fibroblasts and stimulates inositol phosphate formation in L-M and arachidonic acid release in 3T3 cells

Stimulation of murine L-M and 3T3 fibroblasts with human recombinant tumour necrosis factor (rTNF) resulted in an increase in the cytosolic free Ca2+ concentration (Ca2+]i). In 3T3 cells rTNF also induced release and metabolization of arachidonic acid, whereas in L-M cells rTNF provoked rapid increases in the levels of inositol mono-, bis- and trisphosphates (IP1, IP2 and IP3). In these cells the Ca2+ response was also observed in Ca2+ free medium, suggesting that rTNF promotes mobilization of Ca2+ from intracellular stores. In 3T3 cells, however, Ca2+ originated from the extracellular space, since the response was abolished in medium containing 1 mM EGTA. Both rTNF-induced calcium responses were inhibited by a specific rabbit IgG antibody to rTNF but not by 1-verapamil, a blocker potential-operated calcium channels. These results suggest that increased formation of inositol phosphates, arachidonic acid release and increased cytosolic free Ca2+ are involved in the biological effects of rTNF. However, rTNF generate these signals by different mechanisms depending upon the target cell.