Journal: Mediators of Inflammation

Article Title: Analysis of Th17 and Tc17 Frequencies and Antiviral Defenses in Gut-Associated Lymphoid Tissue of Chronic HIV-1 Positive Patients

doi: 10.1155/2015/395484

Figure Lengend Snippet: Activation markers, IFN- γ and IL-17A expression in CD4 + and CD8 + T cells subsets in blood and in gut. Panel (a): representative flow cytometry plots outlining CD38 and HLA-DR expression on naïve, effector, and central memory CD4 + and CD8 + lymphocytes derived from peripheral blood mononuclear cells (PBMCs) and gut-associated lymphoid tissue (GALT). Panel (b): representative flow cytometry plots outlining IFN- γ and IL-17A intracitoplasmatic expression on naïve, effector, and central memory CD4 + (Th1 and Th17) and CD8 + (Tc1 and Tc17) lymphocytes derived from PBMCs and GALT.

Article Snippet: Cultured cells were fixed, permeabilized (BD Cytofix/Cytoperm, Becton Dickinson, San Jose, CA, USA), and stained with combinations of fluorochrome-labeled monoclonal antibodies: CD3-PerCP, CD4-APC-Vio770, CD8-FITC, CD45RO-PE-Vio770, CD27-VioBlue, IFN- γ -APC, and IL-17A-PE (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Activation Assay, Expressing, Flow Cytometry, Derivative Assay

Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: The gene-specific primer pairs list in the study.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques:

Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: Analysis of the mRNA expression of cytokines and transcription factors for Th cells in the colons of TRAF5 KO and WT mice. (a) TNF- α , (b) IFN- γ , (c) IL-4, (d) IL-17a, (e) IL-22, (f) T-bet, (g) GATA-3, (h) ROR- α , and (i) ROR- γ t mRNA expression levels were determined by real-time PCR. The data are representative of 3 independent experiments (mean ± SD). N = 8 per group. ∗ p < 0.05 versus the DSS-treated WT group; ∗∗ p < 0.01 versus the DSS-treated WT group. NS: not significant versus the DSS-treated WT group.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: Deletion of TRAF5 increases the protein expressions of the cytokines IFN- γ , IL-4, and IL-17a in the colons of mice induced by DSS. (a) The production of IFN- γ , IL-4, and IL-17a in the colons of DSS-induced mice or control mice was examined by ELISA ( N = 6 per group). (b) Representative immunofluorescence staining for IFN- γ , IL-4, and IL-17a in the colons of TRAF5 KO and WT mice (magnification 400x). (c) Quantification of positive cells ( N = 4 per group). The data are representative of 3 independent experiments (mean ± SD). ∗ p < 0.05 versus the DSS-treated WT group; ∗∗ p < 0.01 versus the DSS-treated WT group.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

Journal: Mediators of Inflammation

Article Title: Disruption of Tumor Necrosis Factor Receptor-Associated Factor 5 Exacerbates Murine Experimental Colitis via Regulating T Helper Cell-Mediated Inflammation

doi: 10.1155/2016/9453745

Figure Lengend Snippet: Increased proportions of Th2 and IFN- γ /IL-17a-coproducing CD4 + T cells in the colons of TRAF5-deficient animals induced by DSS. LPMCs obtained from the colons of DSS-fed KO and WT mice were stimulated with PMA/ionomycin and subjected to surface staining for CD4. After fixation and permeabilization, intracellular staining for IL-4, IFN- γ , and IL-17a was performed as described in . CD4 + T cells were then gated and analyzed. (a) Representative flow plots. The percentages of (b) Th2 cells (CD4 + IL-4 + ), (c) IFN- γ /IL-17a-coproducing CD4 + T cells (CD4 + IFN- γ + IL-17a + ), (d) classical Th1 cells (CD4 + IFN- γ + IL-17a − ), and (e) classical Th17 cells (CD4 + IL-17a + IFN- γ − ) are presented as the mean ± SD. The data are representative of 3 independent experiments. N = 6 per group. ∗∗ p < 0.01 versus the DSS-treated WT group. NS: not significant versus the DSS-treated WT group.

Article Snippet: The following primary antibodies were used: anti-IFN- γ (1 : 200 dilution; Proteintech, USA), anti-IL-4 (1 : 100 dilution; BioLegend, USA), anti-IL-17a (1 : 200 dilution; Proteintech, USA), anti-p65 (1 : 100 dilution; Cell Signaling Technology, USA), anti-CD4 (1 : 100 dilution; BioLegend, USA), and anti-CK18 (1 : 100 dilution; Goodbio Technology Co., China).

Techniques: Staining

Journal: Molecules (Basel, Switzerland)

Article Title: Inosine Prevents Colorectal Cancer Progression by Inducing M1 Phenotypic Polarization of Macrophages.

doi: 10.3390/molecules30010123

Figure Lengend Snippet: Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of CD86 and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.

Article Snippet: Primary antibodies against CD86 (1:1000, 26903-1-AP, Proteintech, Wuhan, China), iNOS (1:300, 22226-1-AP, Proteintech, Wuhan, China), β-actin (1:2000, GB15003-100, Servicebio, Wuhan, China) and secondary antibody against HRP Goat Anti-Rabbit IgG (1:5000, AS014, ABclonal, Wuhan, China) were used, respectively.

Techniques: MTT Assay, Flow Cytometry, Expressing, Quantitative RT-PCR, Control

Journal: Molecules (Basel, Switzerland)

Article Title: Inosine Prevents Colorectal Cancer Progression by Inducing M1 Phenotypic Polarization of Macrophages.

doi: 10.3390/molecules30010123

Figure Lengend Snippet: Figure 4. Effects of inosine on immune factors in the CT26 tumor microenvironment. (A) Effect of inosine on Ki-67 expression in tumor tissues (Scale: 100 µm; 400× and 200×). (B) Statistics of Ki-67 protein positive expression in tumor tissues (n = 3). (C) Fluorescence co-localization fluorogram of M1- type macrophage marker F4/80 + CD86 (scale: 100 µm; 200×). (D) F4/80 + CD86 expression statistics in tumor tissues. (E) M2 type macrophage marker F4/80 + CD206 fluorescence co-localization fluorogram (scale: 100 µm; 200×). (F) F4/80 + CD206 expression statistics in tumor tissues. Note: 5-Fu are 5-Fu (12 mg/kg) groups. IS-L and IS-H are inosine low and high-dose (5 mg/kg and 50 mg/kg) groups, respectively. Yellow arrows represent positive positions. Compared with the model group: ## p < 0.01; Compared with the 5-Fu group: △p < 0.05.

Article Snippet: Primary antibodies against CD86 (1:1000, 26903-1-AP, Proteintech, Wuhan, China), iNOS (1:300, 22226-1-AP, Proteintech, Wuhan, China), β-actin (1:2000, GB15003-100, Servicebio, Wuhan, China) and secondary antibody against HRP Goat Anti-Rabbit IgG (1:5000, AS014, ABclonal, Wuhan, China) were used, respectively.

Techniques: Expressing, Fluorescence, Marker