Activation of human monocytic cells by lysophosphatidic acid and sphingosine-1-phosphate.

Lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are serum-borne lipid mediators with potential proinflammatory and atherogenic properties. We studied the effects of LPA and S1P on Ca(2+), a second messenger of cellular activation, in human monocytic Mono Mac 6 (MM6) cells. LPA and S1P induced Ca(2+) transients with EC(50) values of 47 and 340 nM, respectively. Ca(2+) signals evoked by LPA and S1P originated mainly from the stimulation of Ca(2+) entry, were blocked by the phospholipase C inhibitor U73122, and were inhibited by pertussis toxin. The LPA(1) and LPA(3) receptor antagonist dioctylglycerol pyrophosphate inhibited the LPA-induced Ca(2+) signal. Notably, serum and minimally modified LDL (mm-LDL) evoked Ca(2+) increases that were mediated entirely through activation of LPA receptors. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis showed the presence of the LPA and S1P receptor subtypes LPA(1), LPA(2,) S1P(1), S1P(2), S1P(4) in MM6 cells, human monocytes and macrophages. Together these results indicate that LPA, mm-LDL and serum induce via activation of the LPA(1) receptor a G(i)/phospholipase C/Ca(2+) signalling pathway in monocytes. Our study is the first report showing the receptor-mediated activation of human monocytic cells by low nanomolar concentrations of LPA and S1P, and suggests a role of these lipid mediators in inflammation and atherogenesis.