Journal: Rheumatology (Oxford, England)

Article Title: Inhibition of CCL3 abrogated precursor cell fusion and bone erosions in human osteoclast cultures and murine collagen-induced arthritis

doi: 10.1093/rheumatology/key196

Figure Lengend Snippet: CCL3 correlates with bone resorption in vitro OCP ( n = 8 donors, four replicates/condition) differentiated into osteoclasts in medium containing anti-CCL3 or IgG1 antibody. ( A ) Anti-CCL3 exerted a concentration-dependent inhibition on osteoclastogenesis (i), reducing total tartrate-resistant acid phosphatase (TRAP)-positive cells (ii), multinucleated osteoclasts (iii) and resorption (iv). ( B ) Disks stained with TRAP and haematoxylin (day 14). M-CSF cultures lacked osteoclasts (top), M-CSF + RANKL cultures showed strong TRAP staining (bottom; scale bar = 50 µm). ( C ) Resorption pits visualized by toluidine blue (top, scale bar = 250 μm) or calcein (bottom) reduced in anti-CCL3 (8 ng/ml) cultures vs IgG1 (yellow arrows). Mean value per donor plotted. * P ≤ 0.05, ** P ≤ 0.01. OCP = osteoclast precursor cells.

Article Snippet: Cell responses were compared using media supplemented with mouse IgG1 isotype as a control (MAB002, R&D Systems, Abingdon, UK).

Techniques: In Vitro, Concentration Assay, Inhibition, Staining

Journal: Rheumatology (Oxford, England)

Article Title: Inhibition of CCL3 abrogated precursor cell fusion and bone erosions in human osteoclast cultures and murine collagen-induced arthritis

doi: 10.1093/rheumatology/key196

Figure Lengend Snippet: CCL3 inhibition had no effect on resorption pit parameters Calcein-stained ivory disks were imaged by fluorescence microscopy to quantify resorption pit parameters. ( A ) Representative topographical maps of M-CSF (i), M-CSF + RANKL + IgG1 (ii), M-CSF + RANKL + anti-CCL3 (iii) disks show their naturally undulating surface and resorption pits (spherical lacuna); scale bar = 40 μm. ( B ) Measured lacuna area (i), perimeter (ii), depth (iii) and volume (iv) for anti-CCL3 vs IgG1 were unchanged (8 ng/ml). ( C ) Levels of CCL2 (i) and sIL-6R (ii) were comparable in IgG1 and anti-CCL3 (8 ng/ml) cultures (day 14). Cells from healthy human volunteers ( n = 6) were cultured, n = ≤2 disks/condition, mean ( s . e . m .) for each donor plotted.

Article Snippet: Cell responses were compared using media supplemented with mouse IgG1 isotype as a control (MAB002, R&D Systems, Abingdon, UK).

Techniques: Inhibition, Staining, Fluorescence, Microscopy, Cell Culture

Journal: Rheumatology (Oxford, England)

Article Title: Inhibition of CCL3 abrogated precursor cell fusion and bone erosions in human osteoclast cultures and murine collagen-induced arthritis

doi: 10.1093/rheumatology/key196

Figure Lengend Snippet: Systemic inhibition of CCL3 reduced histological joint swelling Mice with CIA received IgG1 or anti-CCL3 on days 21, 23, 25, 27 and 28 (5 mg/kg, n = 6/group). ( A ) Arthritis progression monitored by clinical scores (i) and paw diameter (ii) saw no statistical differences (two-way analysis of variance). ( B ) Representative tartrate-resistant acid phosphatase (TRAP) and haematoxylin-stained elbow joints from IgG1 (i) and anti-CCL3 (ii). Assessment of inflammation (iii), erosion (iv) and arthritic index (v). ( C ) IgG1 wrist histology showed intense TRAP-staining (i), which reduced in anti-CCL3 (ii), with significant reductions in inflammation (iii), erosion (iv) and arthritic index (v). H = humerus, U = ulna, R = radius, C = carpal. * P ≤ 0.05, ** P ≤ 0.01, scale = 1 mm.

Article Snippet: Cell responses were compared using media supplemented with mouse IgG1 isotype as a control (MAB002, R&D Systems, Abingdon, UK).

Techniques: Inhibition, Staining

Journal: Rheumatology (Oxford, England)

Article Title: Inhibition of CCL3 abrogated precursor cell fusion and bone erosions in human osteoclast cultures and murine collagen-induced arthritis

doi: 10.1093/rheumatology/key196

Figure Lengend Snippet: Bone erosions decreased after anti-CCL3 treatment during CIA Front and hind paws from CIA mice, treated with IgG1 or anti-CCL3, were processed for histology. ( A ) tartrate-resistant acid phosphatase (TRAP)-positive cells in the elbow (i) and wrist (ii) joints significantly reduced with anti-CCL3. ( B ) Radiographs of hind paws and ( C ) front paws were acquired from mice treated with IgG1 (i) or anti-CCL3 (ii) ( n = 6 mice per group). Significant reductions in erosive radiographic score for in both hind [B (iii)] and front [C (iii)] paws were quantified in mice treated with anti-CCL3 compared with IgG1 controls. * P ≤ 0.05, ** P ≤ 0.01.

Article Snippet: Cell responses were compared using media supplemented with mouse IgG1 isotype as a control (MAB002, R&D Systems, Abingdon, UK).

Techniques:

Journal: Nutrients

Article Title: Oral Tolerance Induction to Newly Introduced Allergen is Favored by a Transforming Growth Factor-β-Enriched Formula

doi: 10.3390/nu11092210

Figure Lengend Snippet: Total IgE ( A ) and OVA-specific IgG1 ( B ) levels in 5-week-old mice subjected to a food allergy protocol. Mice received PBS using a gavage on days 6, 7, and 8 (non-tolerized/water), OVA (10 mg/mL) ad libitum from day 4 to day 8 (tolerized/water), or a sub-tolerogenic dose of OVA (0.5 mg) using a gavage on days 6, 7, and 8 (ST/water). Values represent the median ± interquartile range of six mice for the non-tolerized/water and tolerized/water groups and of five mice for the three ST/water groups. The significant p -values (≤0.05) are shown on the graphs.

Article Snippet: Serially diluted standard monoclonal mouse anti-OVA IgG1 (used as a proxy for Th2 immune response) [ ] (Antibody Shop; LucernaChem, Lucerne, Switzerland) and plasma samples were incubated for 2 h at 37 °C, followed by a 2-h incubation with an horseradish peroxidase (HRP)-labelled goat anti-mouse IgG1 antibody (1/5000; Southern Biotech, Allschwil, Switzerland).

Techniques:

Journal: Nutrients

Article Title: Oral Tolerance Induction to Newly Introduced Allergen is Favored by a Transforming Growth Factor-β-Enriched Formula

doi: 10.3390/nu11092210

Figure Lengend Snippet: Effect of the oral supplementation of mice with a TGF-β-enriched formula on the protection against sensitization and response to oral exposure to OVA. ( A ) Total IgE, ( B ) OVA-specific IgG1, and ( C ) mMCP1 levels in plasma in 5-week-old mice subjected to a food allergy protocol. Mice received PBS using a gavage on days 6, 7, and 8 (non-tolerized/water), OVA (10 mg/mL) ad libitum from day 4 to day 8 (tolerized/water), or a sub-tolerogenic dose of OVA (0.5 mg) using a gavage on days 6, 7, and 8 (ST/water). ST/water mice had free access to either water, control formula, or TGF-β-enriched formula from day 1 to day 13. Values are expressed as the median ± interquartile range of eight mice per group. Significant p -values (≤0.05) are shown on the graphs.

Article Snippet: Serially diluted standard monoclonal mouse anti-OVA IgG1 (used as a proxy for Th2 immune response) [ ] (Antibody Shop; LucernaChem, Lucerne, Switzerland) and plasma samples were incubated for 2 h at 37 °C, followed by a 2-h incubation with an horseradish peroxidase (HRP)-labelled goat anti-mouse IgG1 antibody (1/5000; Southern Biotech, Allschwil, Switzerland).

Techniques: