Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD40/CD40 ligand interactions are critical for elicitation of autoimmune-mediated fibrosis in the lung.

doi: 10.4049/jimmunol.166.5.3556

Figure Lengend Snippet: FIGURE 4. IL-12 and TNF-a ELISA on BAL samples from HIPIF mice treated with anti-CD40L Ab. Histograms show the level of IL-12 (p70) (a), IL-12 (p40/70) (b), and TNF-a (c) in the lavage from experi- mental mice (A, A9, B, C). A, HIPIF mice; A9, HIPIF mice treated with

Article Snippet: Recombinant mouse (rm) IL-12 (p70) was purchased from R&D Systems (Minneapolis, MN).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD40/CD40 ligand interactions are critical for elicitation of autoimmune-mediated fibrosis in the lung.

doi: 10.4049/jimmunol.166.5.3556

Figure Lengend Snippet: FIGURE 8. Hydroxyproline deposition in the lungs of HIPIF mice treated with anti-CD40L Ab and rmIL-12 (p70). The histogram shows the change (D) of hydroxyproline in each experimental group (A, A9, A0, A-, B, B9, and C) 14 days after i.t. challenge. The treatment of each group is indicated under the abscissa. A, HIPIF mice treated with normal Armenian hamster IgG (h IgG, control Ab); A9, HIPIF mice treated with anti-CD40L Ab; A0, HIPIF mice treated with anti-CD40L Ab and rmIL-12 (100 ng/ mouse); A-, HIPIF mice treated with anti-CD40L Ab and rmIL-12 (500 ng/mouse); B, i.t. challenge-only mice treated with h IgG; B9, i.t. chal- lenge-only mice treated with anti-CD40L Ab. The administration of Ab and rmIL-12 is described in Materials and Methods. The change (D) in hydroxyproline was calculated as experimental hydroxyproline 2 baseline hydroxyproline. The baseline hydroxyproline collected from naive mice was 215.8 mg/lung. Five mice were used in each experimental group. The data are presented as mean 6 SEM. An asterisk (p) indicates a statistically significant difference between the two groups indicated.

Article Snippet: Recombinant mouse (rm) IL-12 (p70) was purchased from R&D Systems (Minneapolis, MN).

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Intraclonal Enrichment of IL-23 Receptor Complex Expression in the Proliferative Fraction of Chronic Lymphocytic Leukemia

doi: 10.3390/ijms27031202

Figure Lengend Snippet: IL-23R and IL-12R expression in ex vivo CLL cells assessed by flow cytometry. FSC is proportional to cell size and SSC reflects intracellular complexity/granularity (each dot represents one cell). Axes in marker plots show fluorescence intensity for the indicated markers. Percentages within gates indicate the fraction of cells relative to the parent population. Lymphocytes were identified by FSC/SSC properties, followed by doublet exclusion using pulse height vs. width plots. Within the single-cell gate, CD19 + /CD5 + CLL cells were selected. This population was further sub-divided into RF and PF based on the differential expression of CD184 and CD5. ( A ) Gating strategy used to identify lymphocytes, single cells, CD19 + CLL cells, and to discriminate proliferative fraction (PF) and resting fraction (RF); ( B ) surface expression of IL-23R, IL-12Rβ1, and IL-12Rβ2 receptor subunits in ex vivo CLL cells, shown for CD19 + PF and RF populations; ( C ) representative flow cytometry plots showing sorting of PF and RF populations from six CLL samples according to the gates indicated; ( D ) IL-12Rβ1 mRNA expression in sorted RF and PF cells, measured by RT-qPCR. Data are shown as mean ± SEM. Statistical significance of the difference is evaluated using the two-sided Wilcoxon signed-rank test. * p < 0.05; ** p < 0.01, ns: not significant. FSC: forward scatter; SSC: side scatter; PE Phycoerythrin; PE-Cy7: Phycoerythrin–Cyanine 7; APC: Allophycocyanin; AF 488: Alexa Fluor 488; PE VIO 770: near-infrared fluorochrome emitting in the ~770–780 nm range.

Article Snippet: The following monoclonal antibodies were used: mouse anti human IL-23R-PE (Cat. #FAB14001P, R&D Systems, Minneapolis, MN, USA), mouse anti human IL-12Rβ1-BB515 (CD212, BD Hori-zon, Cat. #565043, BD Biosciences, San Jose, CA, USA, and mouse anti human IL-12Rβ2-PerCP (Cat. #FAB1959C, R&D Systems).

Techniques: Expressing, Ex Vivo, Flow Cytometry, Marker, Fluorescence, Single Cell, Quantitative Proteomics, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Intraclonal Enrichment of IL-23 Receptor Complex Expression in the Proliferative Fraction of Chronic Lymphocytic Leukemia

doi: 10.3390/ijms27031202

Figure Lengend Snippet: IL-23R and IL-12R expression in CLL cells after incubation with CpG and CpG + IL-15 for 72 h assessed by flow cytometry analyses. ( A ) Representative flow cytometry gating strategy used to identify CLL cells and to assess IL-23R and IL-12R complex expression under the indicated culture conditions (medium alone, CpG, or CpG + IL-15) after 72 h; ( B ) percentage of CLL cells expressing the IL-23R receptor subunit, IL-12Rβ1 receptor subunit, and IL-12Rβ2 receptor subunit following 72 h incubation under the indicated conditions; ( C ) percentage of CLL cells expressing the IL-23R complex and the IL-12R complex following 72 h incubation under the indicated conditions. Statistical significance of the difference is evaluated using the two-sided Wilcoxon signed-rank test. * p < 0.05; **** p < 0.0001. FSC: forward scatter; SSC: side scatter; PE Phycoerythrin; PE-Cy7: Phycoerythrin–Cyanine 7; APC: Allophycocyanin; AF 488: Alexa Fluor 488.

Article Snippet: The following monoclonal antibodies were used: mouse anti human IL-23R-PE (Cat. #FAB14001P, R&D Systems, Minneapolis, MN, USA), mouse anti human IL-12Rβ1-BB515 (CD212, BD Hori-zon, Cat. #565043, BD Biosciences, San Jose, CA, USA, and mouse anti human IL-12Rβ2-PerCP (Cat. #FAB1959C, R&D Systems).

Techniques: Expressing, Incubation, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Intraclonal Enrichment of IL-23 Receptor Complex Expression in the Proliferative Fraction of Chronic Lymphocytic Leukemia

doi: 10.3390/ijms27031202

Figure Lengend Snippet: Differential expression of IL-23R and IL-12R in CLL RF and PF subsets after stimulation with CpG and CpG + IL-15 assessed by flow cytometry analyses. ( A ) Representative flow cytometry gating strategy used to identify CLL cells and to distinguish RF and PF subsets, followed by assessment of IL-23R and IL-12R complex expression after 72 h incubation with CpG or CpG + IL-15; ( B ) percentage of RF and PF CLL cells expressing the IL-23R receptor subunit, IL-12Rβ1 receptor subunit, and IL-12Rβ2 receptor subunit after 72 h incubation under the indicated conditions; ( C ) percentage of RF and PF CLL cells expressing the IL-23R complex and the IL-12R complex after 72 h incubation under the indicated conditions. Each symbol represents an individual CLL sample; horizontal bars indicate mean ± SEM. Statistical significance of the difference is evaluated using the two-sided Wilcoxon signed-rank test. ** p < 0.01; **** p < 0.0001, ns: not significant. FSC: forward scatter; SSC: side scatter; PE Phycoerythrin; PE-Cy7: Phycoerythrin–Cyanine 7; APC: Allophycocyanin; AF 488: Alexa Fluor 488; BV421 = Brilliant Violet 421.

Article Snippet: The following monoclonal antibodies were used: mouse anti human IL-23R-PE (Cat. #FAB14001P, R&D Systems, Minneapolis, MN, USA), mouse anti human IL-12Rβ1-BB515 (CD212, BD Hori-zon, Cat. #565043, BD Biosciences, San Jose, CA, USA, and mouse anti human IL-12Rβ2-PerCP (Cat. #FAB1959C, R&D Systems).

Techniques: Quantitative Proteomics, Flow Cytometry, Expressing, Incubation