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. 2021 Sep 10;12:720183. doi: 10.3389/fimmu.2021.720183

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Copyright © 2021 Garam, Cserhalmi, Prohászka, Szilágyi, Veszeli, Szabó, Uzonyi, Iliás, Aigner, Schmidt, Gaggl, Sunder-Plassmann, Bajcsi, Brunner, Dumfarth, Cejka, Flaschberger, Flögelova, Haris, Hartmann, Heilos, Mueller, Rusai, Arbeiter, Hofer, Jakab, Sinkó, Szigeti, Bereczki, Janko, Kelen, Reusz, Szabó, Klenk, Kóbor, Kojc, Knechtelsdorfer, Laganovic, Lungu, Meglic, Rus, Kersnik Levart, Macioniene, Miglinas, Pawłowska, Stompór, Podracka, Rudnicki, Mayer, Rysava, Reiterova, Saraga, Seeman, Zieg, Sládková, Stajic, Szabó, Capitanescu, Stancu, Tisljar, Galesic, Tislér, Vainumäe, Windpessl, Zaoral, Zlatanova, Józsi and Csuka

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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(A) Comparison of serum Factor H-related protein 5 (FHR-5) levels and serum FHR-5 C3b-binding ability of patients with only FHR-5_WT and those carrying FHR-5_G278S or FHR-5_R356H in ELISA. C3b was immobilized, then serum was added, and FHR-5 was detected by polyclonal anti-FHR-5. Dots represent individual patients. Data are mean of two measurements and are normalized to a calibrator sample containing only FHR-5_WT. Subjects expressing FHR-5G278S show a tendency of lower FHR-5 levels and decreased C3b binding compared to the FHR-5WT-expressing group. Serum FHR-5 levels were measured in a sandwich ELISA. Circles represent individual patients. Data are mean of two measurements. (B) Dose-dependent binding of recombinant FHR-5_WT, FHR-5_G278S, and FHR-5_R356H to purified C3b measured in ELISA. The FHR-5_G278S variant binds significantly weaker to C3b (one-way ANOVA with Bonferroni’s posttest, p < 0.01). Data are mean of four measurements ± SEM. (C) Interaction of C3b–FHR-5 variants measured by surface plasmon resonance (SPR). C3b in serial dilutions was flown over immobilized FHR-5 variants. The K_D of the C3b–FHR-5_G278S is one order greater compared to that of C3b–FHR-5_R356H and C3b–FHR-5_WT. Data are representative of two experiments.