Journal: Annals of the rheumatic diseases

Article Title: Cathepsin S inhibition suppresses systemic lupus erythematosus and lupus nephritis because cathepsin S is essential for MHC class II-mediated CD4 T cell and B cell priming.

doi: 10.1136/annrheumdis-2013-203717

Figure Lengend Snippet: Figure 4 R05461111 reduces autoantibodies in MRL-Fas(lpr) mice. (A) Antinuclear antibodies (ANA) staining patterns on Hep2 human epithelial cells for serum derived from vehicle-treated and R05461111-treated MRL-Fas(lpr) mice at 1:200 dilution. (B) Crithidia luciliae slides were incubated with 1:50 diluted plasma of 20-week-old mice from both treatment groups, and autoantibody binding to the flagellate’s kinetoplast was detected using an FITC-labelled goat antimouse IgG. Images on the left show anti-dsDNA IgG in green, and staining the kinetoplast DNA itself with DAPI shown in blue is shown in the middle. The merged pictures are shown on the right side, demonstrating that FITC positivity matches with the kinetoplast at the flagella pole of C luciliae. (C) Plasma levels of anti-dsDNA IgG and IgM antibodies were determined by ELISA. (D) Plasma levels of IgG isotypes such as IgG1, IgG2a, IgG2b and IgG3 against ds-DNA were determined by ELISA. (E) Plasma levels of anti-RNP/Sm, antinucleosome antibodies and rheumatoid factor were determined by ELISA. Data are expressed as means±SEM (n=15 each treatment group,). *p<0.05, **p<0.01, ***p<0.001 versus vehicle.

Article Snippet: For the detection of IgG and its isotypes in the serum, plates were precoated with antimouse IgG, IgG1, IgG2a, IgG2b and IgG3 (Bethyl Laboratories, Montgomery, Texas, USA), and sera were applied at dilutions of 1:10 000 to 1:50 000.

Techniques: Staining, Derivative Assay, Incubation, Clinical Proteomics, Binding Assay, Enzyme-linked Immunosorbent Assay