Journal: Mucosal immunology

Article Title: Regulatory B Cells from Hilar Lymph Nodes of Tolerant Mice in a Murine Model of Allergic Airway Disease are CD5 + , Express TGF-β and Co-localize with CD4 + Foxp3 + T Cells

doi: 10.1038/mi.2012.42

Figure Lengend Snippet: Panel A: CD4 + CD25 − T cells from spleens of naïve mice were co-cultured for 5 days without or with stimulation by soluble anti-CD3 and anti-CD28. Stimulated cells were also co-cultured with either irradiated CD19 + CD5 + B cells or CD19 + CD5 − B cells from LIT HLNs. T cell Foxp3 expression was increased when cultured with LIT HLN CD5 + B cells versus with LIT CD5 − B cells or anti-CD3/CD28 alone. Data represent mean ± SEM values of 3–4 animals per group; ** indicates p < 0.01 vs. other groups by ANOVA. Panel B: Foxp3 + Treg expression was compared in LIT control and JhD −/− mice. Control mice showed expansion of Foxp3 + Treg cells in hilar nodes (black bars) but not inguinal nodes (striped bars), but this local expansion did not occur in the JhD −/− mice. n = 5 mice per group; * indicates p < 0.005 between CRL and JhD −/− mice. Panel C: Two days before a week of daily OVA aerosol exposures, OVA-sensitized mice received tail vein injections of saline or specific B cell populations. Mice receiving LIT HLN CD5 + B cells demonstrated increased numbers of Foxp3 + T cells in their BAL, as compared to control (saline) mice or mice receiving LIT HLN CD5 − B cells or LIT spleen CD5 + B cells. n = 5–11 mice per group; * indicates p < 0.05 vs. other groups by ANOVA. Panel D: For all groups of mice in panel C, there was a direct correlation between the number of CD4 + Foxp3 + Treg cells and the number of CD19 + CD5 + B cells in BAL (dashed line; r = 0.56; p < 0.005). This relationship also held for the LIT HLN CD5 + recipient animals alone (solid line; r = 0.73; p < 0.05). In contrast, there was no association between CD4 + CD25 + Foxp3 − Teff cells and CD19 + CD5 + B cells in BAL (r = 0.28; p > 0.10; data not shown).

Article Snippet: B cells from HLNs and spleens of LIT mice were positively selected using a mouse CD19 + isolation kit (Miltenyi Biotech, Auburn, CA).

Techniques: Cell Culture, Irradiation, Expressing, Control, Aerosol, Saline

Journal: Mucosal immunology

Article Title: Regulatory B Cells from Hilar Lymph Nodes of Tolerant Mice in a Murine Model of Allergic Airway Disease are CD5 + , Express TGF-β and Co-localize with CD4 + Foxp3 + T Cells

doi: 10.1038/mi.2012.42

Figure Lengend Snippet: CD5 + B cells were isolated from hilar nodes (black bars) and inguinal nodes (gray bars) at different stages (Naïve, Sensitized, AAD and LIT) of the OVA model. Panel A depicts representative flow cytometry dot plots of CXCR4 expression on CD19 + CD5 + B cells. Panel B demonstrates increased CXCR4 + CD5 + B cells in hilar compared to inguinal lymph nodes at all stages of the model, and expansion of hilar node CXCR4 + CD5 + B cells during AAD and LIT. Panel C demonstrates similar CXCR5 expression by CD5 + B cells in both tissues and at all stages of the model. Data represent the mean ± SEM; n = 8–12 in each group (A, B); * indicates p < 0.05 as compared to Naïve and Sensitized groups in the HLN and to all groups in the ILN; ! indicates p < 0.05 as compared to all groups in the ILN; † indicates p < 0.05 between HLN and ILN in Naïve and Sensitized groups; †† indicates p < 0.005 between HLN and ILN in AAD and LIT groups.

Article Snippet: B cells from HLNs and spleens of LIT mice were positively selected using a mouse CD19 + isolation kit (Miltenyi Biotech, Auburn, CA).

Techniques: Isolation, Flow Cytometry, Expressing

Journal: Mucosal immunology

Article Title: Regulatory B Cells from Hilar Lymph Nodes of Tolerant Mice in a Murine Model of Allergic Airway Disease are CD5 + , Express TGF-β and Co-localize with CD4 + Foxp3 + T Cells

doi: 10.1038/mi.2012.42

Figure Lengend Snippet: OVA-sensitized mice received tail vein injections of saline or specific B cell populations (0.2 × 10 6 CD19 + CD5 + or 1.0 × 10 6 CD19 + CD5 − cells) 2 days before a week of daily exposure to 1% OVA aerosols. Mice receiving the LIT HLN CD5 + B cells (solid bars) developed attenuated AAD, with less relative (Panel A) and absolute (Panel B) airway eosinophilia relative to control (saline) mice or mice receiving LIT HLN CD5 − B cells or LIT spleen CD5 + B cells. Data represent mean ± SEM values of 5–11 mice per group; * indicates p < 0.05 vs. other groups by ANOVA.

Article Snippet: B cells from HLNs and spleens of LIT mice were positively selected using a mouse CD19 + isolation kit (Miltenyi Biotech, Auburn, CA).

Techniques: Saline, Control

Journal: Cell

Article Title: Lymph Nodes are Innervated by a Unique Population of Sensory Neurons with Immunomodulatory Potential

doi: 10.1016/j.cell.2020.11.028

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Next, using the Miltenyi CD3ε microbead kit and CD19 mouse microbead kit, all remaining LN cells were stained according to manufacturer instructions with the following modifications.

Techniques: Virus, Recombinant, Multiplex Assay, Expressing, Software

Journal: Molecules

Article Title: Sensitization to Drug Treatment in Precursor B-Cell Acute Lymphoblastic Leukemia Is Not Achieved by Stromal NF-κB Inhibition of Cell Adhesion but by Stromal PKC-Dependent Inhibition of ABC Transporters Activity

doi: 10.3390/molecules26175366

Figure Lengend Snippet: B-ALL cells show increased susceptibility to conventional chemotherapy after PKC inhibition in the mesenchymal support. ( A ) Cell viability of primary B-ALL cells after PKC inhibition and treatment with MTX. As control, B-ALL cells without treatment (NT) were included. MSC were treated with HKPS, the commercial inhibitor ENZA, or the controls for 2 h, as indicated. B-ALL cells, pre-treated 6 h with 6.25 µM of MTX, were co-cultured for additional 72 h on MSC. Co-cultures were collected after trypsinization and the cell viability was assessed by flow cytometry on the leukemic cell population through the double staining with the reactive LIVE/DEAD Aqua and the anti-CD19 antibody. A representative experiment is shown. ( B ) Quantification of the B-ALL cell population double positive for CD19 and LIVE/DEAD Aqua as described in A. ( C ) Quantification of the viability of the B-ALL cells after inhibition of PKC in MSC with HKPS, HK (40 µM), ENZA (20 µM), or the vehicle (0.4% DMSO) and co-culturing for 72 h with B-ALL cells pre-treated with VNC (500 nM). ( D ) Simultaneous effect of DEXA and HKPS on B-ALL cells viability. Leukemic cells were pre-treated for 6 h with DEXA (250 nM), HKPS (40 µM), or both, and then they were co-cultured for 24 h with non-treated MSC (CO-CULTURE (MSC NT)); also, co-cultures were established and then cells were treated with the combination of HKPS and DEXA (CO-CULTURE TREATED); as controls, B-ALL cells without support were employed (B-ALL ALONE, upper panel). Cell viability was assessed by flow cytometry. A representative experiment is shown. ( E ) Percentages of B-ALL cells viability in the conditions indicated above. Data are expressed as mean ± SEM ( p values: Non-parametric one-way ANOVA. * p < 0.05. ** p < 0.01. *** p < 0.001) from two independent experiments.

Article Snippet: The cells were incubated with the Gold Dye reactive provided in the Kit and the Mouse anti-human CD19 (clone SJ25C1, BD Pharmingen, San Jose, CA, USA) and mouse anti-human CD73 (clone AD2, BD Pharmingen, San Jose, CA, USA) antibodies for 30 min. Propidium iodide supplied in the kit was added at the end of the incubation period in order to assess the transporters activity in the viable leukemic cell population.

Techniques: Inhibition, Cell Culture, Flow Cytometry, Double Staining, Co-Culture Assay

Journal: Antioxidants

Article Title: Effect of Heme Oxygenase-1 on Melanoma Development in Mice—Role of Tumor-Infiltrating Immune Cells

doi: 10.3390/antiox9121223

Figure Lengend Snippet: Effect of HO-1 expression in mice on leukocytes infiltration in primary tumors. ( A ) Percentage of leukocytes (CD45 + cells) in tumors. Cytometric analysis. Each point represents mean ± SE ( N = 3–9). * p < 0.05 vs. HO-1 +/+ mice, # p < 0.05 ♂ vs. ♀. ( B ) Subpopulations of infiltrating leukocytes: CD11b + Gr1 + myeloid cells, CD11b + Gr1 − myeloid cells, CD3 + T lymphocytes; CD19 + B lymphocytes; Nk 1.1 + NK cells. Cytometric analysis.

Article Snippet: Antibodies against CD3, CD11b, CD19, CD45, Gr-1, and NK 1.1, the BD™ Cytometric Bead Array (CBA) Mouse Inflammation kit and 2,2′,2″,2‴-(Ethane-1,2-diyldinitrilo)tetraacetic acid (EDTA) were purchased from BD Biosciences (San Jose, CA, USA).

Techniques: Expressing

Journal: Antioxidants

Article Title: Effect of Heme Oxygenase-1 on Melanoma Development in Mice—Role of Tumor-Infiltrating Immune Cells

doi: 10.3390/antiox9121223

Figure Lengend Snippet: Effect of HO-1 expression in mice on concentrations of proangiogenic and proinflammatory cytokines in sera after intracutaneous injection with melanoma cells. ( A ) Concentration of IFN-γ. Cytometric analysis. ( B ) Concentration of keratinocyte chemoattractant (KC). ELISA. ( C ) Concentration of IL-10. Cytometric analysis. ( D ) Concentration of IL-6. Cytometric analysis. ( E ) Concentration of vascular endothelial growth factor (VEGF). Cytometric analysis. ( F ) Concentration of IL-12. Cytometric analysis. ( G ) Concentration of MCP-1. ELISA. Each bar represents mean ± SE ( N = 4–10; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. HO-1 +/+ mice, # p < 0.05, ## p < 0.01♂ vs. ♀).

Article Snippet: Antibodies against CD3, CD11b, CD19, CD45, Gr-1, and NK 1.1, the BD™ Cytometric Bead Array (CBA) Mouse Inflammation kit and 2,2′,2″,2‴-(Ethane-1,2-diyldinitrilo)tetraacetic acid (EDTA) were purchased from BD Biosciences (San Jose, CA, USA).

Techniques: Expressing, Injection, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Antioxidants

Article Title: Effect of Heme Oxygenase-1 on Melanoma Development in Mice—Role of Tumor-Infiltrating Immune Cells

doi: 10.3390/antiox9121223

Figure Lengend Snippet: Effect of HO-1 expression in mice on concentrations of proangiogenic and proinflammatory cytokines in sera after intravenous injection with melanoma cells. ( A ) Concentration of IFN-γ. Cytometric analysis. ( B ) Concentration of KC. ELISA. ( C ) Concentration of VEGF. ELISA. ( D ) Concentration of IL-10. Cytometric analysis. ( E ) Concentration of IL-12. Cytometric analysis. ( F ) Concentration of MCP-1. Cytometric analysis. ( G ) Concentration of IL-6. Cytometric analysis. Each bar represents mean ± SE ( N = 3–8; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. HO-1 +/+ mice, # p < 0.05, ♂ vs. ♀).

Article Snippet: Antibodies against CD3, CD11b, CD19, CD45, Gr-1, and NK 1.1, the BD™ Cytometric Bead Array (CBA) Mouse Inflammation kit and 2,2′,2″,2‴-(Ethane-1,2-diyldinitrilo)tetraacetic acid (EDTA) were purchased from BD Biosciences (San Jose, CA, USA).

Techniques: Expressing, Injection, Concentration Assay, Enzyme-linked Immunosorbent Assay