C-reactive protein and natural IgM antibodies are activators of complement in a rat model of intestinal ischemia and reperfusion.

BACKGROUND: The role of C-reactive protein (CRP), natural immunoglobulin M (IgM), and natural IgM against phosphorylcholine (anti-Pc IgM) was investigated in relation with complement activation in a rat model of intestinal ischemia and reperfusion (II/R). The effect of C1-esterase inhibitor (C1-Inh) on this complement activation along with other inflammatory mediators was also studied. METHODS: Rats were subjected to 1 h of superior mesenteric artery occlusion and 3 h of reperfusion. Intravenous administration of vehicle (human albumin) or C1-Inh (200 U/kg) was performed before (n = 8) or after ischemia (n = 8). II/R increased levels of C4b/c, CRP, IgM, anti-Pc IgM, and myeloperoxidase activity in the intestinal homogenates and induced vascular leakage. RESULTS: A good correlation was observed in the intestinal homogenates between C4b/c and CRP levels. Clear depositions of C3, CRP, and IgM in intestinal tissue were demonstrated after II/R, and a strong correlation of both CRP and IgM with complement was observed. C1-Inh administered before ischemia reduced the complement activation response after II/R, as reflected by decreased levels of C4b/c in conjunction with reduced anti-Pc IgM in the intestinal homogenates. C1-Inh also decreased leakage of albumin when administered before ischemia. C1-Inh after ischemia reduced C4b/c levels and myeloperoxidase activity in the homogenates. CONCLUSIONS: CRP and IgM depositions correlated well with local complement activation, which suggests a role of these molecules in complement activation. Furthermore, C1-Inh inhibited potentially II/R injury either administered before or after ischemia, by attenuating complement activation induced by CRP and/or natural IgM antibodies.

Levels of natural IgM antibodies against phosphorylcholine in healthy individuals and in patients undergoing isolated limb perfusion.

Natural IgM antibodies against phosphorylcholine (anti-Pc IgM) resemble C-reactive protein (CRP) regarding specificity and have gained increasing attention because of their supposed role in clearance of damaged cells and in cardiovascular disease. In order to quantify these antibodies in human plasma, we have developed an ELISA system, in which p-aminophenylphosphorylcholine (PCH) coupled to human serum albumin (HSA) was coated on microtiters plates. Human plasma or serum samples were incubated in the plates, after which bound anti-Pc IgM was detected with mouse anti-human IgM-HRP. Pre-incubation of plasma with competitors such as phosphorylcholine, phosphorylethanolamine, phosphorylserine or glycine-HSA, confirmed that the ELISA was specific for anti PC IgM. Levels of anti Pc IgM in a cohort of healthy donors differed by more than 100-fold, whereas the fluctuation of anti-Pc IgM levels in individuals over time was small (coefficient of variation between 6% to 25%). Furthermore, there was no correlation between CRP and anti-Pc IgM in this cohort. Levels of anti-Pc IgM in the normal donors correlated significantly with IgM binding to apoptotic cells. To test the hypothesis that anti-Pc IgM can bind to neo-antigens expressed on necrotic or apoptotic cells, anti-Pc IgM was also quantified in patients with tumors undergoing isolated limb perfusion with tumor necrosis factor-alpha (TNF-alpha). Following this procedure, a significant decrease of circulating anti-Pc IgM relative to total IgM was found in all five patients tested. In conclusion, we have developed a specific and reproducible ELISA for anti Pc IgM quantification. Fluctuation of levels of these natural antibodies over time in healthy individuals was limited, although the variation among individuals was large. Significant decreases of levels of anti-Pc IgM were found to occur during tissue damage.