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: Fig. 3 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: Fig. 5 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

Journal: American journal of physiology. Renal physiology

Article Title: Inhibition of T-cell activation by the CTLA4-Fc Abatacept is sufficient to ameliorate proteinuric kidney disease.

doi: 10.1152/ajprenal.00179.2016

Figure Lengend Snippet: Fig. 5. A: validation of anti-human B7-1 antibodies. Of all 4 commercial antibodies tested, we identified an antibody from Novus Biologics (Novus 62N3G8) that recognized a diffuse, 65-kDa band in T-cells consistent with the glycosylated estate of B7-1 and its predicted molecular weight (MW) of 65 kDa. This antibody also recognized recombinant human B7-1 although yielding a slightly lower MW. This could be due to intrinsic modifications of the protein introduced to facilitate the purification process. B: absence of B7-1 protein in podocytes with and without LPS and HG treatment. Conditionally immortalized human podocytes were differentiated at 37°C for at least 14 days and were used for experiments in passages between 5 and 20.

Article Snippet: Anti-human B7-1 antibodies used were Novus Biologicals 62N3G8 (Western blotting) and Abcam ab134120 (IHC).

Techniques: Biomarker Discovery, Molecular Weight, Recombinant

Journal: Mediators of Inflammation

Article Title: Increased Migratory and Activation Cell Markers of Peripheral Blood Lymphocytes in an Experimental Model of Nephrotic Syndrome

doi: 10.1155/2015/209764

Figure Lengend Snippet: CD18 and CD80 expression in total monocytes from rats with nephrotic syndrome induced by doxorubicin and control animals in ex vivo condition. Blood leukocytesfrom rats ( n = 32) that received a single dose of doxorubicin in tail vein (7.5 mg/kg) and control rats ( n = 32) that received PBS in the same condition were stained ex vivo for CD18 and CD80 expression. Data were collected using flow cytometry and analyzed using CellQuest software. Graphs (a) MFI of CD18 + monocytes and (b) MFI of CD80 + monocytes show expression of the given marker in lymphocytes. The data expressed as mean ± standard error for Dox group (black bars) and control group (grew bars). ∗ p < 0.05 for the comparison between Dox and control group at the same time-point (Student's t -test).

Article Snippet: Whole peripheral blood (50 μ L/sample) was incubated with monoclonal antibody for the following surface markers: anti-CD3, anti-CD4, anti-CD8 (Becton & Dickinson, San Jose, CA, USA), anti-CD80, and anti-CD18 (Caltag-Medsystems Limited, Buckingham, UK) conjugated with fluorescein phycoerythrin (PE), isothiocyanate (FITC), or biotin in the dark for 30 minutes at room temperature.

Techniques: Expressing, Ex Vivo, Staining, Flow Cytometry, Software, Marker

Journal: Mediators of Inflammation

Article Title: Increased Migratory and Activation Cell Markers of Peripheral Blood Lymphocytes in an Experimental Model of Nephrotic Syndrome

doi: 10.1155/2015/209764

Figure Lengend Snippet: Correlations of redox status in kidney tissue, renal function parameters, and immune markers in rats with nephrotic syndrome induced by doxorubicin.

Article Snippet: Whole peripheral blood (50 μ L/sample) was incubated with monoclonal antibody for the following surface markers: anti-CD3, anti-CD4, anti-CD8 (Becton & Dickinson, San Jose, CA, USA), anti-CD80, and anti-CD18 (Caltag-Medsystems Limited, Buckingham, UK) conjugated with fluorescein phycoerythrin (PE), isothiocyanate (FITC), or biotin in the dark for 30 minutes at room temperature.

Techniques:

Journal: Oncotarget

Article Title: Carbon ion irradiation enhances the antitumor efficacy of dual immune checkpoint blockade therapy both for local and distant sites in murine osteosarcoma

doi: 10.18632/oncotarget.26551

Figure Lengend Snippet: (A) Changes in PD-L1 protein expression by flow cytometry. (B) Representative flow cytometric plots of PD-L1 expression. (C) Changes in CD80 protein expression by flow cytometry. (D) Representative flow cytometric plots of CD80 changes. Error bars show standard deviation. P-values were determined by Tukey's honestly significant difference tests for the comparison between each dose level. Bars represent P<0.01.

Article Snippet: At 3 days after irradiation, cells were reacted with anti-PD-L1-PE antibody (Clone: MIH5, BD Pharmagen, NJ, USA) and anti-CD80-FITC antibody (Clone: 16-10A1, BD Pharmagen, NJ, USA).

Techniques: Expressing, Flow Cytometry, Standard Deviation, Comparison