Journal:

Article Title: CTLA-4 regulates allergen response by modulating GATA-3 protein level per cell

doi: 10.1111/j.1365-2567.2007.02537.x

Figure Lengend Snippet: Effects of in vitro CTLA-4 blockade on GATA-3 protein level/cell and frequency of IL-4-producing cells. Naive CD4+ T cells purified from the spleens of normal mice were induced to differentiate with anti-CD3 mAb, a recombinant form of CD80-Fc, IL-4 and anti-IFN-γ mAb for 3 days. Anti-CTLA-4 mAb was added in soluble form at increasing concentrations (0·1–10 μg/ml). (a) Mean fluorescence intensity for GATA-3 staining, (b) percentage of IL-4+ cells.

Article Snippet: 18 CD4 cell culture For in vitro experiments, splenic naive CD4 cells were purified from non-immunized BALB/c mice by immuno-magnetic cell sorting as previously described 19 and cultured in 24-well plates precoated with anti-CD3ε mAb (clone 145-2C11; 10 μg/ml) and/or recombinant mouse B7-1(CD80)/Fc chimeric protein (R & D Systems, cat. 740-B1; 0·33 μg/ml).

Techniques: In Vitro, Purification, Recombinant, Fluorescence, Staining

Journal: BMC Immunology

Article Title: TRPV1 + neurons alter Staphylococcus aureus skin infection outcomes by affecting macrophage polarization and neutrophil recruitment

doi: 10.1186/s12865-023-00584-x

Figure Lengend Snippet: TRPV1 + neurons affect the polarization of local macrophages in the skin. a IHC staining of the skin of TRPV1 −/− and WT mice; scale bars, 500 μm; high magnification images, scale bars, 50 μm. b - d IHC analysis of the ratio of M0 (F4/80 +), M1 (CD80 +) and M2 (CD206 +) cells in TRPV1 −/− and WT mice ( n = 5/group); unpaired t test. e – h Flow cytometric analysis of M0 (CD11b + F4/80 +), M1 (CD11b + F4/80 + CD80 +), and M2 (CD11b + F4/80 + CD206 +) ratios in the skin of TRPV1 −/− and WT mice ( n = 5/group); unpaired t test. i - k Tissue expression levels of TNFα, IL-1β and IL-10 in TRPV1 −/− and WT mice 1 day after skin infection; the control group was uninfected WT mice; One-way ANOVA with Tukey’s post hoc test. Data were pooled from two or three independent experiments

Article Snippet: After washing, the cells were fixed/permeabilized according to the instructions provided by the manufacturer (MultiSciences, FoxP3/Transcription Factor Staining Buffer Kit) and then incubated on ice with CD206 (Elabscience, #E-AB-F0992C) for 30 min. Flow cytometry data were collected and exported using Beckman CytoFLEX (USA).

Techniques: Immunohistochemistry, Expressing, Infection, Control

Journal: BMC Immunology

Article Title: TRPV1 + neurons alter Staphylococcus aureus skin infection outcomes by affecting macrophage polarization and neutrophil recruitment

doi: 10.1186/s12865-023-00584-x

Figure Lengend Snippet: CGRP regulates the polarization of BMDMs and the release of inflammatory factors. a - c BMDMs cultured in vitro were induced to M1 polarization with IFN-γ and M2 polarization with IL-4 The polarized macrophages were treated with CGRP or PBS and stained with DAPI (blue), CD80 (green), and CD206 (red); the ratio of CD80 + and CD206 + was analyzed under different intervention conditions. No difference in the confluency of BMDMs was observed among different groups. Scale bars, 20 μm. One-way ANOVA with Tukey’s post hoc test. d - f Expression levels of TNFα, IL-1β, and IL-10 in BMDMs after polarization in vitro under different experimental conditions; one-way ANOVA with Tukey posttests. Data were pooled from two or three independent experiments

Article Snippet: After washing, the cells were fixed/permeabilized according to the instructions provided by the manufacturer (MultiSciences, FoxP3/Transcription Factor Staining Buffer Kit) and then incubated on ice with CD206 (Elabscience, #E-AB-F0992C) for 30 min. Flow cytometry data were collected and exported using Beckman CytoFLEX (USA).

Techniques: Cell Culture, In Vitro, Staining, Expressing