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. 2017 Mar 14;1(8):504–515. doi: 10.1182/bloodadvances.2016001917

Figure 6.

Figure 6.

GL-2045 promotes the regulatory function of factors H and I and induces enhanced iC3b generation. (A) Self-limited complement activation by GL-2045 and IVIG is dependent on factor H. Factor H–depleted serum was incubated with various concentrations of GL-2045, HAGG, and IVIG for 90 minutes at 37°C. The sera were then evaluated for the presence of C4a, C3a, and C5a. (B) GL-2045 inhibits the activity of the alternative form of C3 convertase activity in the presence of factor H. GL-2045, HAGG, or IVIG was added to the alternative C3 convertase assembly at indicated concentrations in the presence or absence of factor H (25 μg/mL). C3 cleavage was measured by C3a ELISA. (C) In a different set of experiments, fractions of GL-2045 were added to the alternative C3 convertase assembly in the presence of factor H and C3. C3a generation was evaluated. (D) GL-2045 works in concert with the combination of factors H and I to inhibit the function of alternative form of C3 convertase. The alternative C3 convertase was generated in the presence of combination of GL-2045 and factors H and I. (E) Levels of iC3b were significantly increased in NHS treated with GL-2045. NHS was exposed to GL-2045, HAGG, or IVIG for 90 minutes at 37°C. The levels of iC3b in the serum were determined by ELISA. The results are shown as the least squares mean ± SE estimated by ANCOVA. Natural log variance stabilizing transformation and Tukey procedure (A,E), square root variance stabilizing transformation and Dunnett’s procedure (B), or log variance stabilizing transformation and Dunnett procedure (C-D) for multiple comparisons adjustment were used to test the differences. *P < .05, **P < .01, ***P < .001 compared with no-treatment (A,E) or vehicle (B-D) control.