Journal: Molecular therapy. Nucleic acids
Article Title: Systemic RNAi-mediated Gene Silencing in Nonhuman Primate and Rodent Myeloid Cells
doi: 10.1038/mtna.2011.3
Figure Lengend Snippet: Inhibition of collagen mAb-induced arthritis by small interfering RNA (siRNA) against tumor necrosis factor- α (TNF α ) . ( a ) Preventative treatment of mice with siRNA against TNFα (purple curve) but not against Luc (blue curve) in C12-200 formulation decreases arthritis score. Anti-VLA1 mAb given several hours before arthitogenic antibody mix was used as a positive control (green curve) as previously shown. Mice were treated with anticollagen mAb's at day 0, followed by lipopolysaccharide (LPS) on day 3. Mice receiving only arthitogenic antibodies are graphed in red. 0.5 mg/kg siRNA in C12-200 formulation was administered on days −3, 0, 4, and 7. All mice were scored on days 4–8 and 10. Each limb was evaluated and scored on a 0–4 scale. Results are expressed as the mean arthritic score (±SEM) of all four limbs (maximum score of 16). Groups of 10 mice per condition were used; one out of two experiments is shown, both showed similar protection. ( b ) Intensity of intracellular TNFα staining in CD11b + splenic macrophages from diseased mice at day 10. Groups of three mice were analyzed; bars represent average of the mean fluorescent intensity in the group with SEM, statistical analysis as described in the Results section. ( c ) Detection of inflammatory macrophage activity by fluorescence-mediated tomography (FMT) imaging. Arthritic mice treated with siRNAs targeted to TNFα or Luc were injected with Prosense-750, a pan-specific cathepsin substrate, and imaged by FMT on day 7. Fluorescence concentration reports on protease activity in the joint and was quantified as described in Supplementary Materials and Methods , n = 5, 2 paws per mouse were analyzed. Data are mean ± SEM. ( d ) Joint histology at 5x magnification in H&E staining correlated with images of the paws. ( e ) Representative images of macrophage activity measurement by FMT quantified in panel c . Naive and siRNA-treated animals are shown.
Article Snippet: Arthrogen-collagen–induced arthritis antibody kits were purchased from Chondrex LLC (Redmond, WA), and arthritis was induced using an established protocol., Groups of 10 BALB/c mice (CRL) were injected intraperitoneally with a cocktail of five anticollagen type II mAb's (0.75 mg/animal total, Chondrex, catalog #53100) on day 0, followed by intraperitoneal injection of 25 µg lipopolysaccharide on day 3.
Techniques: Inhibition, Small Interfering RNA, Positive Control, Staining, Activity Assay, Fluorescence, Tomography, Imaging, Injection, Concentration Assay
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