In contrast to the phorbol ester oxidative response, which only develops during dimethyl‐sulphoxide (DMSO)‐induced differentiation of the human leukemic myeloblast HL‐60 cell‐line, the endotoxin response was observed in undifferentiated and differentiated cells. The Ca2+ response to endotoxin, detected in both differentiated and undifferentiated HL‐60 cells, consisted of a transient 10–50 nM increase in intracellular Ca2+. A very slow, irreversible increase in intracellular Ca2+ was detected at high 1–100 μg/ml endotoxin concentrations, and this effect, and the inositol phosphate response, correlated with the surfactant activities of various endotoxins and Lipid A. Arachidonic acid and sodium arachidonate 1–50 μM stimulated a large 200–500 nM and transient Ca2+ response in undifferentiated HL‐60 cells, which was significantly greater than that elicited by 1–50 μM eicosapentaenoic acid, and was not observed at similar concentrations of arachidonic acid methyl ester or myristic acid. These concentrations (1–50 μM) of arachidonic acid were observed to have surfactant activities on the plasma membrane. At lower arachidonic acid concentrations a marked potentiation of both Ca2+ and oxidative responses to the chemotactic peptide fMet‐Leu‐Phe was detected. It is possible that the arachidonic acid released during phospholipase A2 activation of neutrophils may be involved in cellular cross‐talk and, at higher concentrations, in directly activating Ca2+ and superoxide production. It is also possible that previously reported effects of endotoxin at high concentrations are an vitro artefact of surfactant properties of endotoxin.