Figure 2.

Treatment with anti-CD20 reduces glomerular complement activation in mice with adriamycin nephropathy. Mice are injected with anti-CD20 mAb to deplete B cells before the induction of adriamycin nephropathy. Complement activation in the glomeruli is examined by immunofluorescence microscopy 4 weeks after injection with adriamycin. (A) Staining for C4 demonstrates that glomerular C4 deposition increases after injection with adriamycin, but that treatment of the mice with anti-CD20 prevents this increase. (B) Staining for C3 demonstrates that glomerular C3 deposition increases after injection with adriamycin. Treatment of the mice with anti-CD20 prevents this increase in C3 deposition. Glomeruli are indicated with arrowheads and areas of C3 and C4 staining within the glomeruli are indicated with small arrows. Kidneys from three to five mice per group are examined. Thirty glomeruli per section are visualized, and the average for each mouse is determined. After converting the images to grayscale, the brightness and contrast of the shown images are adjusted. All images are adjusted equally. (C) Dual staining of kidneys from mice with adriamycin nephropathy for IgM and C3 demonstrates colocalization of IgM and C3 within glomeruli (appearing yellow in the overlay). Glomeruli are indicated with arrowheads and areas of colocalization are indicated with small arrows. (D) Staining of kidneys from mice with adriamycin nephropathy for C3d confirms that the glomerular deposits contain activated complement proteins. Original magnification, ×200 in A–C; ×400 in D. Scale bar, 100 μM.