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Blood, Vol. 110, Issue 5, 1516-1518, September 1, 2007
Anti–factor H autoantibodies block C-terminal recognition function of factor H in hemolytic uremic syndrome
Blood Józsi et al.
110: 1516
Supplemental materials for: Jozsi et al, Vol 110, Issue 5, 1516-1518
Files in this Data Supplement:
- Figure S1. Identification of autoantibody positive patients (JPG, 62.3 KB)
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(A) Plasma samples of aHUS (n = 51) and D+ HUS (n = 9) patients as well as healthy donors (n = 30) were analyzed for binding on immobilized purified FH in an ELISA assay. The samples having an OD value above the mean plus 2 SD of those in the control group were considered positive. (B) Dose-dependent binding of the autoantibody-positive plasma samples of 5 aHUS patients to immobilized FH, indicating different autoantibody titers. (C) Adsorption of IgG from the patients’ plasma by incubation with Protein G beads removed FH autoreactivity. (D) Isotypes of FH-specific IgG in the plasma samples were determined using mouse antibodies specific to human IgG1, IgG2, IgG3, and IgG4.
- Figure S2. Competition assays show C-terminal specificity for autoantibody binding (JPG, 81.8 KB)
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C-terminally binding mAbs (C02, C14, and C18; dotted lines), but not mAbs binding outside SCR19-20 (continuous lines), exhibited dose-dependent inhibitory activity on autoantibody binding to FH as shown for plasma of (A) patient no. 564 and (B) patient no. 520. (C) The FH SCR15-20 fragment (dotted lines), but not the SCR15-18 fragment (continuous lines), inhibited autoantibody binding to FH in a dose-dependent manner as shown for three patients.
- Figure S3. Characterization of the IgG fraction purified from patient’s plasma (JPG, 31.5 KB)
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(A) Purified IgG of patient no. 564 bound specifically to the FH fragments SCR15-20 and SCR19-20 (ie, those containing the most C-terminal domains; filled bars). Control IgG purified from a healthy donor showed no specific binding (open bars). (B) Dose-dependent binding of purified IgG from patients no. 422, no. 520, and no. 564 to immobilized FH.
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