Journal: European Journal of Trauma and Emergency Surgery

Article Title: Isolated blunt chest injury leads to transient activation of circulating neutrophils

doi: 10.1007/s00068-010-0041-x

Figure Lengend Snippet: Expression of l -selectin ( a ), αM (CD11b) ( b ), CXCR1 ( c ), CXCR2 ( d ), and C5aR ( e ) on the neutrophil surface over time. Open squares indicate control values from healthy controls ( N = 8), whereas black squares represent patients ( N = 13) at 3, 9 and 24 h postinjury. C , control; hash symbol , time of injury. Data are presented as mean ± SE ( *p < 0.05, **p < 0.01)

Article Snippet: The following commercially available mouse–antihuman monoclonal antibodies were purchased for analyzing neutrophil receptor expression by flowcytometry: fluorescein isothiocyanate (FITC)-labeled IgG1 isotype control (clone MOPC-21, BD Pharmingen, USA), Alexa Fluor ® 647-labeled IgG1 isotype control (clone MOPC-21, BD Pharmingen, USA), R-phycoerythrin (RPE)-labeled IgG2a isotype control (clone MRC OX-34, Serotec, Germany), RPE-labeled IgG1 anti-αM (CD11b; clone 2LPM19c, DAKO, Denmark), FITC-labeled IgG1 anti- l -selectin (CD62L; clone Dreg56, BD Pharmingen, USA), Alexa Fluor ® 647-labeled IgG1 anti-FCγRIII (CD16; clone 3G8, BD Pharmingen, USA), RPE-labeled IgG2b anti-FcγRII (CD32; clone FLI8.26, BD Pharmingen, USA), FITC-labeled IgG2a anti-CXCR1 (CD181a; clone 42705, R&D Systems Europe, UK), RPE-labeled IgG2a anti-CXCR2 (CD182b; clone 48311, R&D Systems Europe, UK), and FITC-labeled IgG2a anti-C5aR (CD88; clone P12/1, Serotec, Germany).

Techniques: Expressing

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

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

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: Histologic analysis of synovial tissue sections from juvenile idiopathic arthritis (JIA) patients. Serial synovial tissue sections from three JIA patients were stained with CD20 (a) , CD138 (b) , CD21 (c) , and CD27 (d) mAbs by using the peroxidase method. CD20 + B cells cluster within lymphoid aggregates, whereas CD138 + plasma cells localize at the periphery of such aggregates. Staining for CD21, a follicular dendritic cell marker, is negative, consistent with the absence of follicular organization. CD27 + cells are found both inside and outside the lymphoid aggregates.

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: Staining, Marker

Journal: The Journal of Immunology Author Choice

Article Title: Mucosa-Associated Invariant T Cell Hypersensitivity to Staphylococcus aureus Leukocidin ED and Its Modulation by Activation

doi: 10.4049/jimmunol.2100912

Figure Lengend Snippet: LukED targets mature CD56dim NK cells mostly independent of CCR5. (A) Representative flow cytometry plots and combined donor data of the percentage and absolute count of CD56bright and CD56dim NK cells on LukED treatment for 4 h (n = 6). LukED is used at 5 µg/ml throughout the figure. (B) Percentage of depletion of CD56bright and CD56dim NK cells on LukED treatment (n = 6). Representative flow cytometry plots (C) and combined data (D) of the expression of CXCR1, CXCR2, and CCR5 on CD56bright and CD56dim NK cells (n = 6). (E) Representative flow cytometry plots and combined data of the expression of CD27 and NKG2A on CD56bright and the expression of KIRD2L1, CD57, Perforin, and CD16 on CD56dim NK cells with or without LukED treatment (n = 6). (F) Combined data of the percentage of coexpression of the indicated markers on CD56bright (left) and CD56dim (right) NK cells with or without LukED exposure (n = 6). The Wilcoxon signed-rank test was performed to determine significance. The lines and error bars represent mean and SE. *p < 0.05.

Article Snippet: Abs used included anti-CD3 Bv650 (OKT3; BioLegend), anti-CD3 AF700 (UCHT1; BD Biosciences), anti-CD3 FITC (SK7; BD Biosciences), anti-Vα7.2 PE (3C10; BioLegend), anti-Vα7.2 PE-Cy7 (3C10; BioLegend), anti-CD161 Pe-Cy5 (DX12; BD Biosciences), anti-CD4 Bv711 (OKT4; BioLegend), anti-CD8 Bv570 (RPA-T8; BioLegend), anti-CD56 BUV737 (NCAM16.2; BD Biosciences), anti-CCR7 Bv421 (150503; BD Biosciences), anti-CD45RA AF700 (HI100; BD Biosciences), anti-CCR5 BUV395 (2D7/CCR5; BD Biosciences), anti-CXCR1 AF488 (8F1/CXCR1; BD Biosciences), anti-CXCR2 Bv785 (6C6; BD Biosciences), anti-CD158b Bv510 (CH-L; BD Biosciences), anti-CD16 Bv711 (3G8; BD Biosciences), anti-CD57 PE-Cy5 (NK-1; BD Biosciences), anti-CD27 PE-Cy7 (M-T271; BioLegend), anti-NKG2A allophycocyanin (REA110; Miltenyi), anti-CD14 allophycocyanin-Cy7 (MѻP9; BD Biosciences), anti-CD19 allophycocyanin-Cy7 (SJ27C1; BD Biosciences), anti-Vα24 PE (C15; Beckman Coulter), anti-Vβ11 FITC (C21; Beckman Coulter), anti-γδ TCR PE-Dazzle 594 (B1; BioLegend), anti-CD69 BUV737 (FN50; BD Biosciences), anti-GzmB FITC (GB11; BioLegend), anti–IFN-γ allophycocyanin (25723.11; BD Biosciences), anti-TNF PE-Cy7 (Mab11; BD Biosciences), anti–IL-17A Bv421 (BL168; BioLegend), anti-Perforin Bv421 (B-D48; BioLegend), and anti-Granulysin (DH2; BD Biosciences) LIVE/DEAD Fixable Near-IR dye (Invitrogen).

Techniques: Flow Cytometry, Expressing