Journal: Frontiers in Immunology

Article Title: Comparative immunohistochemical analysis of inflammatory cytokines in distinct subtypes of Sweet syndrome

doi: 10.3389/fimmu.2024.1355681

Figure Lengend Snippet: Cytokine expressions in adult-onset immunodeficiency (AOID)-associated Sweet syndrome (SS) is maintained independently despite the presence of anti-interferon (IFN)-γ autoantibodies. Representative images of hematoxylin and eosin (H&E) staining (top panel) in lesion of AOID- and non-AOID-associated SS and control normal tissues Bar = 1 mm. Immunohistochemical (IHC) staining and the corresponding annotated whole slide images for IFN-γ (middle panel) and interleukin (IL)-17 (bottom panel) in lesion of AOID- and non-AOID-associated SS and controls. Bar = 1 mm. A color markup overlay produced by the color deconvolution algorithm reflecting the intensity ranges of image pixels. Blue, negative; yellow, weak positive; orange, medium positive; red, strong positive.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-human IL-1β [2H12] (sc130323; Santa Cruz Biotechnology, Inc., USA, 1:50 dilution), rabbit polyclonal anti-human IL-6 (ab6672; Abcam, Cambridge, MA, USA, 1:100 dilution), rabbit polyclonal anti-human IL-17A (ab79056; Abcam, 1:100 dilution), rabbit monoclonal anti-human IFN-γ (ab218426; Abcam, 1:50 dilution), and mouse monoclonal anti-human TNF-α [Clone 28401] (mab610, R&D Systems, Minneapolis, MN, USA, 1:100 dilution).

Techniques: Staining, Immunohistochemical staining, Immunohistochemistry, Produced

Journal: Frontiers in Immunology

Article Title: Comparative immunohistochemical analysis of inflammatory cytokines in distinct subtypes of Sweet syndrome

doi: 10.3389/fimmu.2024.1355681

Figure Lengend Snippet: Interferon (IFN)-γ and interleukin (IL)-17 are involved in Sweet syndrome (SS) immunopathology. Graphical representation of percentage of positive (%positivity) and quantification of staining intensity using ( I n+ I wp+ I p+ I sp)/Ntotal data of IFN-γ and IL-17 among adult-onset immunodeficiency (AOID)- and non-AOID-associated SS and the control normal tissues (A) , with or without non-tuberculous mycobacteria (NTM) and control group (B) , and with various forms of non-AOID-associated SS (C) . Data represent the mean ± SD. P values were determined by one-way ANOVA with Tukey adjustments for multiple comparisons where appropriate. Results are expressed in mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. I n, total intensity of negative; I p, intensity threshold of medium-positive pixels; I sp, intensity threshold of strong-positive pixels; I wp, intensity threshold of weak-positive pixels; Ntotal, number of total pixels.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-human IL-1β [2H12] (sc130323; Santa Cruz Biotechnology, Inc., USA, 1:50 dilution), rabbit polyclonal anti-human IL-6 (ab6672; Abcam, Cambridge, MA, USA, 1:100 dilution), rabbit polyclonal anti-human IL-17A (ab79056; Abcam, 1:100 dilution), rabbit monoclonal anti-human IFN-γ (ab218426; Abcam, 1:50 dilution), and mouse monoclonal anti-human TNF-α [Clone 28401] (mab610, R&D Systems, Minneapolis, MN, USA, 1:100 dilution).

Techniques: Staining

Journal: Frontiers in Immunology

Article Title: Comparative immunohistochemical analysis of inflammatory cytokines in distinct subtypes of Sweet syndrome

doi: 10.3389/fimmu.2024.1355681

Figure Lengend Snippet: Schematic representation of the proposed cytokine network implicated in the pathogenesis of Sweet syndrome (SS). The pathogenesis of SS is related to both dysregulated innate and adaptive immune responses, which contribute to the aberration of neutrophil functions. T helper (Th) 1 and Th17 responses may be particularly important in the pathogenesis of SS, as shown by the elevated expression of interferon (IFN)-γ and interleukin (IL)-17 in different subtypes of SS. Because IFN-γ and IL-17 could enhance neutrophil infiltration at sites of inflammation and in turn, neutrophils can amplify inflammatory responses though secreting these cytokines contributing to local inflammation. The expression of IFN-γ, despite the presence of anti-IFN-γ autoantibodies in SS lesions indicates an imbalance of immune system homeostasis.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-human IL-1β [2H12] (sc130323; Santa Cruz Biotechnology, Inc., USA, 1:50 dilution), rabbit polyclonal anti-human IL-6 (ab6672; Abcam, Cambridge, MA, USA, 1:100 dilution), rabbit polyclonal anti-human IL-17A (ab79056; Abcam, 1:100 dilution), rabbit monoclonal anti-human IFN-γ (ab218426; Abcam, 1:50 dilution), and mouse monoclonal anti-human TNF-α [Clone 28401] (mab610, R&D Systems, Minneapolis, MN, USA, 1:100 dilution).

Techniques: Expressing

Journal: Cancers

Article Title: Role of IL-17A and IL-17RA in Prostate Cancer with Lymph Nodes Metastasis: Expression Patterns and Clinical Significance.

doi: 10.3390/cancers15184578

Figure Lengend Snippet: Figure 1. Comparison of IL-17A expression levels in the prostate and metastatic lymph nodes. (a) High IL-17A expression in prostate tissue; (b) Low IL-17A expression in prostate tissue; (c) High IL-17A expression in metastatic lymph node; (d) Low IL-17A expression in metastatic lymph node. Magnification, ×15.

Article Snippet: Endogenous peroxidase was blocked using the EnVision FLEX Peroxidase-Blocking Reagent (Dako) for 5 min. Primary antibodies—polyclonal rabbit antiIL-17/IL-17A antibody (1:1600, cat. no NBP1-76337, Novus Biologicals, Minneapolis, MN, USA) and monoclonal mouse anti-IL17RA/IL-17R antibody (1:200, cat. No NBP2-25258, Novus Biologicals)—were applied for 20 min.

Techniques: Comparison, Expressing

Journal: Neural Regeneration Research

Article Title: Fingolimod protects against neurovascular unit injury in a rat model of focal cerebral ischemia/reperfusion injury

doi: 10.4103/1673-5374.353500

Figure Lengend Snippet: Fingolimod (FTY-720) attenuates ischemia/reperfusion-induced glial cell activation and IL-17A expression. (A) Representative immunofluorescence staining for IL-17A (red) and GFAP (green). (B) Representative immunofluorescence staining for IL-17A (red) and Iba-1 (green). Scale bars: 50 μm. (C) Representative western blot bands for IL-17A. (D) Quantification of IL-17A protein expression ( n = 5 rats per group, * P < 0.05, vs . DMSO group, one-way analysis of variance followed by Bonferroni post hoc test). The experiment was repeated five times. DMSO: Dimethyl sulfoxide; GFAP: glial fibrillary acidic protein; Iba1: ionized calcium-binding adapter molecule 1; IL: interleukin.

Article Snippet: The membranes were blocked for 1 hour with 5% skimmed milk (Solarbio Science & Technology Co., Ltd.) in TBST (tris-buffered saline and Polysorbate 20, also known as Tween 20) at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: rabbit anti-occludin antibody (tight junction protein, 1:1000; Cat# 40-4700, Thermo Fisher Scientific), rabbit anti-claudin-5 polyclonal antibody (tight junction protein, 1:1000; Cat# PA5-99415, Thermo Fisher Scientific), rabbit anti-S1PR1 polyclonal antibody (endothelial cell receptor; 1:1000; Cat# ab11424, Abcam), rabbit anti-IL-17A polyclonal antibody (an inflammatory cytokine; 1:1000; Cat# PA5-106856, Thermo Fisher Scientific), mouse anti-β-actin polyclonal antibody (1:3000; Cat# 3700, CST).

Techniques: Activation Assay, Expressing, Immunofluorescence, Staining, Western Blot, Binding Assay