INHIBITION OF MACROPHAGE NITRIC-OXIDE PRODUCTION AND IA-EXPRESSION BYDOCOSAHEXAENOIC ACID, A CONSTITUENT OF FETAL AND NEONATAL SERUM

PROBLEM: We previously demonstrated profound inhibition of macrophage activation in the murine placenta in vivo. Given the importance of mac rophages both in initiating cellular immunity by presenting antigen in the context of Ia to CD4+ T cells, and in killing cellular targets by producing nitric oxide (NO), inhibition of macrophage functions in th e placenta may account for the increased susceptibility of the placent a to infection. We have also showed that docosahexaenoic acid (DHA), a t concentrations present in the fetal circulation, has a major role in inhibiting macrophage Ia-expression and NO production in the placenta . The concentration of DHA in fetal serum perfusing the placenta is 50 x higher than in the adult. DHA has previously been reported to profou ndly affect prostanoid production, to be metabolized by lipoxygenases, and to affect lipoxygenases. We now determine if these activities of DHA account for its inhibition of macrophage NO production and Ia-expr ession. METHODS: Murine macrophages were cultured in vitro, exposed to IFN gamma, endotoxin, DHA, and various eicosanoids, and their ability to produce NO or express Ia determined. RESULTS: Although the cycloox ygenase inhibitor, indomethacin, did inhibit NO production, DHA inhibi ted by a different mechanism. DHA further inhibited NO production by m acrophages exposed to doses of indomethacin known to maximally inhibit prostanoid production. Stable, biologically active prostanoids did no t reverse the inhibitory effect of DHA. Although DHA is metabolized by lipoxygenases, the lipoxygenase inhibitor NDGA did not reverse the in hibition of either NO production nor Ia expression. This indicates tha t lipoxygenase products of DHA did not mediate inhibition. NDGA itself inhibited NO production and Ia expression. However, DHA did not inhib it by inhibiting lipoxygenase activity because DHA further inhibited m acrophages exposed to doses of DHA known to maximally inhibit lipoxyge nases. Furthermore, stable biologically active analogs of lipoxygenase products did not reverse DHA inhibition. DHA also did not inhibit by preventing PAF production because PAF did not reverse inhibition of NO production. CONCLUSION: DHA did not inhibit Ia-expression or NO produ ction via its known effects on eicosanoid or PAF metabolism, nor by be ing metabolized by lipoxygenases.