Complement activation on solid surfaces as determined by C3 deposition and hemolytic consumption.

Abstract

Complement activation was investigated on hydrophilic and hydrophobic glass beads incubated in serum. Very little complement activation was detected with these surfaces, as indicated by a hemolytic assay and by measurement of the amount of iC3b appearing in the solution. However, preadsorption with IgG at the hydrophobic and hydrophilic beads resulted in complement activation on both surfaces. We also investigated dependent deposition of C3 at hydrophobic and hydrophilic silicon surfaces when the complement was activated. The chemistry of those surfaces is similar to the hydrophobic and hydrophilic beads. Ellipsometry, an optical method, was used for determination of the amounts of organic material deposited at the surface. C3 deposition was observed at the IgG precoated hydrophobic surface but not at the IgG-coated hydrophilic surface. The absence of C3 deposition at the hydrophilic surface was probably due to reversible binding of IgG. However, precoating of the hydrophilic surface with a double layer of IgG and anti-IgG resulted in C3 deposition also at the hydrophilic surface. The results illustrate that methods based on measuring deposition of C3 at surfaces, such as immunofluorescence or ellipsometry, cannot exclude surface-associated complement activation that is probably due to reversible binding of the complement components of the activated molecule. On the other hand, it has previously been shown that determination of complement activation in solution with the use of a C3a assay cannot exclude surface-associated activation due to immobilization of C3a at the surface. This methodologic question is an important issue because factors such as C3a and C5a act as soluble anaphylatoxins, whereas deposited factors such as C3 act as cellular receptors.