Journal: Arthritis Research & Therapy

Article Title: Phenotypic and functional characterization of switch memory B cells from patients with oligoarticular juvenile idiopathic arthritis

doi: 10.1186/ar2824

Figure Lengend Snippet: Flow cytometry of B-cell subsets in synovial fluid (SF) and peripheral blood (PB) from juvenile idiopathic arthritis (JIA) patients. Mononuclear cells (MNCs) from SF and PB of 25 JIA patients, as well as from PB of 20 age-matched controls, were stained with different B cell-specific antibodies and analyzed with flow cytometry. The first gate was set on CD3 - , CD14 - , CD16 - , and CD56 - cells (non-B cell lineage cells) followed by gating on CD19 + cells. Results are expressed in a box plot (left panel) as median percentage of positive cells, minimum and maximum value. * P < 0.01. One representative dot-blot for SF (middle panel) and PB (right panel) is shown.

Article Snippet: The following monoclonal antibodies (mAbs) were used: CD19-phycoerythrin(PE)-cyanin(Cy)7, CD38-PerCP/Cy5, CD27-PerCP/Cy5 from Beckman Coulter (Marseille, France); CD3-allophycocyanin (APC)-Cy7, CD14-APC-Cy7, CD56-biotin and CD16-APC-Cy7, streptavidin-APC-Cy7, CD10-PE; CD24 fluorescein isothiocyanate (FITC), CD20-PE, CD27-PE, CD27-FITC, CD80-FITC; CD86-PE from BD Pharmingen (San Diego, CA, USA); CD10-FITC from Biolegend (San Diego, CA, USA); PE-conjugated anti-human IgD mAb from Dako (Glostrup, Denmark); anti-human immunoglobulin (Ig)G, IgA, and IgM-allophycocyanin (APC) from Jackson Immunoresearch Laboratories (West Grove, PA, USA); PE-conjugated anti-CC chemokine receptor CCR1-CCR9 mAbs from R&D Systems Inc. (Minneapolis, MN, USA); unconjugated anti-CXC chemokine receptor CXCR1, CXCR2, and CXCR3 mAbs from Serotec Inc. (Raleigh, NC, USA); and PE-conjugated anti-CXCR4 and CXCR5 mAbs from R&D Systems.

Techniques: Flow Cytometry, Staining, Dot Blot

Journal: Arthritis Research & Therapy

Article Title: Phenotypic and functional characterization of switch memory B cells from patients with oligoarticular juvenile idiopathic arthritis

doi: 10.1186/ar2824

Figure Lengend Snippet: Transitional B cells in synovial fluid (SF) and peripheral blood (PB) in juvenile idiopathic arthritis (JIA) patients. Cells from SF and PB of JIA patients, as well as from control PB, were analyzed with flow cytometry by gating on CD3 - , CD14 - , CD16 - , and CD56 - cells (non-B cell lineage cells) followed by gating on CD19 + cells and then on CD24 high , CD38 high cells. Results are expressed in a box plot (left panel) as median percentage of positive cells, minimum and maximum value. * P < 0.01 (patient PB versus SF) and P = 0.05 (patient versus control PB). One representative dot-blot for SF (middle panel) and PB (right panel) is shown. Insets on the right side of the Figure show that gated CD24 high and CD38 high B cells express immunoglobulin M and immunoglobulin D (upper inset) and CD10 (lower inset).

Article Snippet: The following monoclonal antibodies (mAbs) were used: CD19-phycoerythrin(PE)-cyanin(Cy)7, CD38-PerCP/Cy5, CD27-PerCP/Cy5 from Beckman Coulter (Marseille, France); CD3-allophycocyanin (APC)-Cy7, CD14-APC-Cy7, CD56-biotin and CD16-APC-Cy7, streptavidin-APC-Cy7, CD10-PE; CD24 fluorescein isothiocyanate (FITC), CD20-PE, CD27-PE, CD27-FITC, CD80-FITC; CD86-PE from BD Pharmingen (San Diego, CA, USA); CD10-FITC from Biolegend (San Diego, CA, USA); PE-conjugated anti-human IgD mAb from Dako (Glostrup, Denmark); anti-human immunoglobulin (Ig)G, IgA, and IgM-allophycocyanin (APC) from Jackson Immunoresearch Laboratories (West Grove, PA, USA); PE-conjugated anti-CC chemokine receptor CCR1-CCR9 mAbs from R&D Systems Inc. (Minneapolis, MN, USA); unconjugated anti-CXC chemokine receptor CXCR1, CXCR2, and CXCR3 mAbs from Serotec Inc. (Raleigh, NC, USA); and PE-conjugated anti-CXCR4 and CXCR5 mAbs from R&D Systems.

Techniques: Flow Cytometry, Dot Blot

Journal: Arthritis Research & Therapy

Article Title: Phenotypic and functional characterization of switch memory B cells from patients with oligoarticular juvenile idiopathic arthritis

doi: 10.1186/ar2824

Figure Lengend Snippet: Plasma blasts and Ig-secreting cells in synovial fluid (SF), peripheral blood (PB), and synovial tissue from juvenile idiopathic arthritis (JIA) patients. (a) Cells from JIA SF and PB, as well as from control PB, were analyzed with flow-cytometry gating, first on CD3 - , CD14 - , CD16 - , and CD56 - cells (non-B cell lineage cells), and then on CD19 + cells, and finally analyzed for CD27 and CD20 expression. Results are expressed in a box plot as median percentage of positive cells, minimum and maximum value. * P < 0.05. One representative dot-blot for SF (middle panel) and PB (right panel) is shown. (b) IgG, IgA, or IgM CD19 + immunoglobulin-secreting cells (ISCs) were detected in SF and PB from four JIA patients with ELISPOT. Results are expressed as mean ISC ± SD. * P = 0.028. (c) Serial synovial tissue sections from three JIA patients were stained with anti-IgG, anti-IgA, or anti-IgM mAbs by using the peroxidase method (brown staining).

Article Snippet: The following monoclonal antibodies (mAbs) were used: CD19-phycoerythrin(PE)-cyanin(Cy)7, CD38-PerCP/Cy5, CD27-PerCP/Cy5 from Beckman Coulter (Marseille, France); CD3-allophycocyanin (APC)-Cy7, CD14-APC-Cy7, CD56-biotin and CD16-APC-Cy7, streptavidin-APC-Cy7, CD10-PE; CD24 fluorescein isothiocyanate (FITC), CD20-PE, CD27-PE, CD27-FITC, CD80-FITC; CD86-PE from BD Pharmingen (San Diego, CA, USA); CD10-FITC from Biolegend (San Diego, CA, USA); PE-conjugated anti-human IgD mAb from Dako (Glostrup, Denmark); anti-human immunoglobulin (Ig)G, IgA, and IgM-allophycocyanin (APC) from Jackson Immunoresearch Laboratories (West Grove, PA, USA); PE-conjugated anti-CC chemokine receptor CCR1-CCR9 mAbs from R&D Systems Inc. (Minneapolis, MN, USA); unconjugated anti-CXC chemokine receptor CXCR1, CXCR2, and CXCR3 mAbs from Serotec Inc. (Raleigh, NC, USA); and PE-conjugated anti-CXCR4 and CXCR5 mAbs from R&D Systems.

Techniques: Flow Cytometry, Expressing, Dot Blot, Enzyme-linked Immunospot, Staining

Journal: Arthritis Research & Therapy

Article Title: Phenotypic and functional characterization of switch memory B cells from patients with oligoarticular juvenile idiopathic arthritis

doi: 10.1186/ar2824

Figure Lengend Snippet: Chemokine receptor and CD86 expression on switch memory B cells from juvenile idiopathic arthritis (JIA) patients. Cells from JIA synovial fluid (SF) and peripheral blood (PB) were analyzed with flow-cytometry gating first on CD3 - , CD14 - , CD16 - , and CD56 - cells (non-B cell lineage cells), then on CD19 + cells, and subsequently on CD27 + or CD27 - cells before being analyzed for individual chemokines-receptor expression. (a) CC chemokine receptor expression on SF and PB CD27 + switch memory B cells. Results are expressed in a box plot as median percentage of positive cells, minimum and maximum value. *** P = 0.0002; P = 0.0003. ** P = 0.001; P = 0.004. * P = 0.019. (b) CXC chemokine receptor and CD86 expression on SF and PB CD27 + switch memory B cells. Results are expressed in a box plot as median percentage of positive cells, minimum and maximum value. *** P = 0.0001. ** P = 0.001; P = 0.002. (c) CCR expression on SF and paired PB CD27 - switch memory B cells. Results are expressed in a box plot as median percentage of positive cells, minimum and maximum value. *** P = 0.0001; P = 0.0003. ** P = 0.0034; P = 0.0041. (d) CXCR and CD86 expression on SF and paired PB CD27 - switch memory B cells. Results are expressed in a box plot as median percentage of positive cells, minimum and maximum value. *** P = 0.0001; P = 0.0006. ** P = 0.0011.

Article Snippet: The following monoclonal antibodies (mAbs) were used: CD19-phycoerythrin(PE)-cyanin(Cy)7, CD38-PerCP/Cy5, CD27-PerCP/Cy5 from Beckman Coulter (Marseille, France); CD3-allophycocyanin (APC)-Cy7, CD14-APC-Cy7, CD56-biotin and CD16-APC-Cy7, streptavidin-APC-Cy7, CD10-PE; CD24 fluorescein isothiocyanate (FITC), CD20-PE, CD27-PE, CD27-FITC, CD80-FITC; CD86-PE from BD Pharmingen (San Diego, CA, USA); CD10-FITC from Biolegend (San Diego, CA, USA); PE-conjugated anti-human IgD mAb from Dako (Glostrup, Denmark); anti-human immunoglobulin (Ig)G, IgA, and IgM-allophycocyanin (APC) from Jackson Immunoresearch Laboratories (West Grove, PA, USA); PE-conjugated anti-CC chemokine receptor CCR1-CCR9 mAbs from R&D Systems Inc. (Minneapolis, MN, USA); unconjugated anti-CXC chemokine receptor CXCR1, CXCR2, and CXCR3 mAbs from Serotec Inc. (Raleigh, NC, USA); and PE-conjugated anti-CXCR4 and CXCR5 mAbs from R&D Systems.

Techniques: Expressing, Flow Cytometry