PBMCs from GPA patients form large TRAP+ MNGs and degrade bone matrix.

<p>PBMCs from both GPA patients and healthy controls were cultured in the presence of 25 ng/ml M-CSF and 100 ng/ml RANKL, and stained for the presence of TRAP expression after 9 days in culture. Nuclei were counterstained with hematoxylin. MNGs with three or more nuclei and TRAP expression (purple cytoplasmic stain) were counted. Few MNGs were generated at day 9 from the PBMCs of healthy controls (A); in contrast, large MNG with numerous nuclei were formed from patients with GPA (representative field shown in B). Further, the generated cells were able to degrade bone matrix. Numerous large pits were formed by GPA PBMCs (D) as compared to the healthy control PBMCs after 9 days (C). C and D are representative of 3 independent experiments. Original magnification x 10.</p

Lack of correlation is noted between MNG formation and circulating monocyte frequency in GPA.

<p>PBMC of healthy controls (N = 10), localized (N = 5) and systemic GPA (N = 8) were stained with anti-human CD14 antibodies and the percentage of CD14+ monocytes were determined using FACS. The number of the circulating monocytes did not differ between three groups (A). Further, percentage of CD14+ monocytes were plotted against formed MNGs per 2×10⁵ PBMC. There was no correlation between percentage of CD14+ monocytes in PBMCs and formed MNGs in both localized and systemic GPA (r² = 0.19, P = 0.46 for localized GPA; r² = 0.17, P = 0.30 for systemic GPA), indicating that systemic GPA formed higher number of MNGs per equivalent number of circulating CD14+ monocytes (B).</p

Disease activity as measured by BASDAI and serum CRP of the two treatment groups during the follow up.

Error bars indicate 95% confidential intervals.</p

Representative photo of negatively-stained HDL₂ from RA patients and controls (electron microscopy).

All micrographs are shown at a magnification of 40,000×. The scale bar corresponds to 100 nm. Graphs show measured particle size and number from designated area. CM, control male; RAM, rheumatoid arthritis male; CF, control female; RAF, rheumatoid arthritis female.</p

HDL₃ associated cholesteryl ester transfer protein (CETP) activity (A) and paraoxonase (PON) activity (B).

CM, control male; RAM, rheumatoid arthritis male; CF, control female; RAF, rheumatoid arthritis female.</p

Uptake of LDL from each group into macrophages was visualized by fluorospectroscopy to detect NBD-cholesterol.

<p>CM, control male; RAM, rheumatoid arthritis male; CF, control female; RAF, rheumatoid arthritis female.</p

Glycation extent of HDL based on fluorescence determination (A) and electrophoretic profiles of HDL as visualized by Coomassie Brilliant Blue staining (B).

<p>CM, control male; RAM, rheumatoid arthritis male; CF, control female; RAF, rheumatoid arthritis female.</p

Increased MNG formation in patients with systemic GPA compared to those with limited disease.

<p>PBMC of healthy controls (N = 11) and GPA patients (N = 13) were cultured in presence of M-CSF and RANKL for 9 days and MNGs counted as in Materials and Methods. PBMCs from GPA patients generated significantly more TRAP+ MNGs than healthy controls (P = 0.022). When the GPA group was analyzed based on disease phenotype [ENT-localized (N = 5) vs. systemic form (N = 8)], only patients with systemic disease formed significantly more MNGs (P = 0.0015); no difference was seen between localized GPA and health controls (P = 0.955).</p

Characteristics of GPA patients.

<p>Yrs, years; F, female; M, male; BVAS-WG, Birmingham Vasculitis Activity Score-Wegener’s Granulomatosis; TRAP, tartrate-resistant acid phosphatase; MNG, multi-nucleated giant cells; AZA, Azathioprine; P, Prednisone; RTX, Rituximab, CTX, cyclophosphamide; MMP, mycophenolate mofetil.</p

Radiographic progressions of the two treatment groups in patients without baseline syndesmophytes (A) and with baseline syndesmophytes (B).

<p>Missing data was replaced using multiple imputation. Values were given as means (standard error) and adjusted for baseline mSASSS. Error bars indicate 95% confidential intervals; mSASSS, modified Stoke Ankylosing Spondylitis Spine Score.</p