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. 2018 Jul 23;9:1691. doi: 10.3389/fimmu.2018.01691

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Copyright © 2018 Zwarthoff, Berends, Mol, Ruyken, Aerts, Józsi, de Haas, Rooijakkers and Gorham.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Development of an alternative pathway (AP) C3 convertase model. (A) C3a specifically induces calcium mobilization in U937-C3aR cells, while C3 and C5 do not. (B) C3 conversion by AP convertases on beads was analyzed in a calcium mobilization assay with U937-C3aR cells. C3a could only be detected in the sample supernatant in the presence of all AP components. C5 conversion in the AP model did not induce calcium flux in the U937-C3aR cells. (C) AP C3 and C5 conversion were performed on beads coated with different densities of C3b and analyzed by calcium mobilization assay with U937-C3aR and -C5aR cells, respectively. A high density of C3b molecules on the target surface enhances C5 but not C3 conversion. (A–C) Data of three independent experiments, presented as mean ± SD.