Article Figures & Data
Figures
- Figure 1 AQP4-IgG–Positive Serum Samples Show Stronger Complement Activation Than MOG-IgG–Positive Serum Samples
(A) CDC (shown as percentage of lysis buffer determined by the LDH assay) of 68 MOG-IgG–positive human serum samples (blue circles; MOGα1-EGFP–transfected cells) and 39 AQP4-IgG–positive human serum samples (red triangles; AQP4-M23-EmGFP–transfected cells) is plotted against the antibody titers. Semi-log linear regression lines are shown. Spearman correlation: MOG: r = 0.58, p < 0.0001; AQP4: r = 0.71, p < 0.0001. (B) Violin plot (dashed line: median, dotted lines: quartiles) of all MOG-IgG (blue; n = 68) and AQP4-IgG (red; n = 39) positive serum samples tested is shown. Asterisks indicate the statistical difference of a Mann-Whitney test: ****p < 0.0001. The solid lines indicate the cutoff levels (mean + 2SD of cells treated with complement alone). (C) Percentage of MOG-IgG (blue) or AQP4-IgG (red) positive samples above the cutoff level (mean + 2SD of cells treated with complement alone) is demonstrated as bar graph. (D) The predictive role of MOG-IgG and AQP4-IgG antibody titers and status, age, and sex on complement-dependent cell lysis were analyzed by multivariate regression (R = 0.827, R2 = 0.684, F = 53.1, p < 0.001). Standardized regression coefficient beta is shown with 95% CI. ***p < 0.001. AQP4 = aquaporin-4; CDC = complement-dependent cytotoxicity; LB = lysis buffer; MOG = myelin oligodendrocyte glycoprotein.
- Figure 2 Complement Activation on Different MOG Isoforms
(A) Comparison of CDC (as percentage of lysis buffer) of MOG-IgG–positive serum samples (n = 68) with 6 different MOG isoforms (MOGα1-3 and MOGβ1-3) is shown as a scatter dot plot. Bars indicate medians with interquartile range (Friedman test with post hoc Dunn: *p < 0.05, ***p < 0.001, and ****p < 0.0001). The solid line indicates the cutoff level (mean + 2SD of cells treated with complement alone). (B) Shows the percentage of MOG-IgG–positive serum samples above the cutoff value (mean + 2SD of cells treated with complement alone) for the 6 MOG isoforms. The percentages are plotted according to their respective binding patterns (α1β1, α1-3β1, and α1-3β1-3; values are given within the bars). AQP4 = aquaporin-4; LB = lysis buffer; MOG = myelin oligodendrocyte glycoprotein.
- Figure 3 Quantification of TCC Deposition
TCC deposition after complement activation of either MOGα1-EGFP (blue) or AQP4-EmGFP (red) transfected HEK293A cells by MOG-IgG or AQP4-IgG–positive serum samples together with human complement was quantified using an flow cytometry assay. (A) Mean fluorescence intensity (MFI) values are plotted against the antibody titers. The lines show semi-log linear regression. Spearman correlation: MOG: r = 0.56, p < 0.05, n = 19; AQP4: r = 0.77, p < 0.001, n = 19. (B) Comparison of MFI values between MOG (n = 19) and AQP4 (n = 19) is shown as violin plot (dashed line: median, dotted lines: quartiles). Mann-Whitney test: **p = 0.002. (C) The MFI values of TCC deposition of human sera are plotted against the respective values from the LDH assay (see Figure 1). Spearman correlation: MOG: r = 0.48, p < 0.05, n = 19; AQP4: r = 0.57, p < 0.05, n = 19. AQP4 = aquaporin-4; LDH = lactate dehydrogenase; MFI = mean fluorescence intensity; MOG = myelin oligodendrocyte glycoprotein; TCC = terminal complement complex.
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