Fig 1: LAIR-1 mediates suppression of CIA. Two groups (n = 10 per group) of DR1/LAIR-1+/+ mice and two groups of littermate DR1/LAIR-1−/− mice were immunized with type II collagen/CFA. Then, each mouse was administered intraperitoneally a dose of either 100µg CI or PBS weekly for four weeks beginning one week after immunization. Each mouse was scored three times weekly beginning 2 weeks post immunization. The severity of the arthritis was statistically significant comparing CI treatment with PBS in LAIR-1+/+ mice beginning on day 44 (p ≤ 0.05) post immunization and continuing until the end of the experiment on day 56. (p ≤ 0.001).
Fig 2: Phosphorylation of human LAIR1 by citrullinated collagen. Jurkat cells that overexpress the human LAIR1 (hLAIR1) were stimulated with PBS, collagen (200 µg/mL), or citrullinated collagen (200 µg/mL) for 5 min. Human LAIR1 was immunoprecipitated from 500 µg of total proteins by anti-human LAIR1 antibody (α-hLAIR1 (NKTA255), Thermo Fisher) using protein A/G bead. Phosphorylation of human LAIR1 was examined with the anti-phosphotyrosine antibody (α -pTyr 4G10, Millipore Human LAIR1 was used as control. The left panel represents type I collagen, and the right panel represents type II collagen. Data are representative of three separate analyses.
Fig 3: Cit-Collagen binds to LAIR with similar affinity as native Collagen. Wells were coated with CI, cit-CI, CII, or cit-CII before adding recombinant LAIR-1 (murine LAIR-1 in the (upper panel) and human LAIR-1 in the (lower panel)) at the indicated concentrations. Binding was detected using an HRP conjugated α -murine LAIR-1 ab, or an α-human LAIR-1 ab followed by a biotinylated α IgG and streptavidin -HRP with reading of absorbance at 450. The control protein, ovalbulmin did not bind the collagens murine LAIR-1 or human LAIR-1.
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