We have previously reported that heterologous, homologous and autologous sera, all stimulated rabbit alveolar macrophages to synthesize prostaglandins (PG). Gel permeation chromatography of serum showed multiple fractions possessing this stimulatory activity, with the major one at 150-160K daltons. In the present study, we have shown that: (a) Fresh rabbit serum stimulated PG release by macrophages. (b) Serum depleted of C3 and C5 lost its stimulatory activity. (c) Trypsinized serum, sera activated by aggregated IgG and zymosan, partially purified C3, C5 and the C3, C5 preparation or purified C3 activated by zymosan, all stimulated PG release by macrophages with the following order of potency: activated C3, C5 = activated C3 = zymosan-activated serum greater than trypsinized serum = aggregated IgG-activated serum greater than partially purified C3, C5 = serum. PGE2 was the predominant PG synthesized by stimulated macrophages. However, thromboxane (TX) production seemed to be more selectively enhanced i.e., increase in TX production was more pronounced than the increase in PGE release. To further identify the active complement component, we blocked the C3b receptor (C3 b R) by preincubating macrophages with anti-C3bR, and showed that subsequent treatment with activated C3 and C5 failed to elicit any PG release. This pretreatment with anti-C3bR had no inhibitory effect on subsequent zymosan stimulation of PG release. Thus we concluded that C3b was the major serum protein that stimulates PG synthesis by macrophages.