Reversal of Lupus Nephritis by SAP Treatment Figure 1. ALD-DNA immunization induces high levels of anti-dsDNA antibody and lupus nephritis. (A) Schematic diagram of animal immunization. 6- to 8-week old female BALB/c mice were immunized subcutaneously with ALD-DNA (50 mg/mice) plus CFA at week 0, followed by two booster immunizations of ALD-DNA (50 mg/mice) emulsified with IFA at week 2 and week 4 after initial immunization. (B) Serum anti-dsDNA IgG levels were measured by ELISA every 2 weeks after initial immunization. Data are means 6 SD from 10 mice in each group. (C) Urine protein levels of the mice were assessed by BCA Protein Assay Kit every 2 weeks. Data are means 6 SD from 10 mice in each group. (D) 8 weeks after initial immunization, nephritic pathology was evaluated by H&E staining of renal tissues. Imagines (magnification6200) are representative of at least 10 mice in each group. (E) The kidney score was assessed using paraffin sections stained with H&E in (D). ***, p,0.001. (F) Glomerular immune deposition were detected by direct immunofluorescence for IgG in frozen kidney section from ALD-DNA-immunized lupus mice or control mice. Representative images (magnification6200) of 10 mice are shown for each group. (G) Mean glomerular fluorescence intensity (arbitrary units) was determined for IgG in ALD-DNA-immunized lupus mice (n = 10) and control mice (n = 10). ***, p,0.001. doi:10.1371/journal.pone.0022659.g001 PLoS ONE | www.plosone.org 3 July 2011 | Volume 6 | Issue 7 | e22659