Journal: Immunology letters

Article Title: Increased expression of membrane-bound TGF-β1, GITR, and GITR ligand in patients with autoimmune thyroid disease.

doi: 10.1016/j.imlet.2025.107036

Figure Lengend Snippet: Fig. 4. Immunohistochemical staining of thyroid tissue from healthy subjects (A and D), AITD patient with GD (B and E), and HD (C and F) with anti-GITRL antibody (D, E, and F) and isotype controls (A, B, and C). GITRL is visualized using diaminobenzidine (brown) and counterstained with hematoxylin. GITRL expression is depicted on thyroid follicular cells. Arrows indicate GITRL expression on infiltrating cells. Magnification ×400.

Article Snippet: For the working dilution, the mouse anti-GITRL polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was diluted at 1:150.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Immunology letters

Article Title: Increased expression of membrane-bound TGF-β1, GITR, and GITR ligand in patients with autoimmune thyroid disease.

doi: 10.1016/j.imlet.2025.107036

Figure Lengend Snippet: Fig. 5. Histoscores of GITRL expression on thyroid follicular cells from patients with AITD (GD [black circle] and HD [white circle]) and healthy subjects. Wilcoxon test is used to compare differences in histoscores between AITD pa tients and healthy subjects.

Article Snippet: For the working dilution, the mouse anti-GITRL polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was diluted at 1:150.

Techniques: Expressing

Journal: Immunology letters

Article Title: Increased expression of membrane-bound TGF-β1, GITR, and GITR ligand in patients with autoimmune thyroid disease.

doi: 10.1016/j.imlet.2025.107036

Figure Lengend Snippet: Fig. 6. Serum titers of (A) TgAb or (B) McAb in AITD patients (GD [black circle] and HD [white circle]) with each histoscore of GITRL expression on thyroid follicular cells. Mann–Whitney U test is used to compare differences in TgAb or McAb between patients with AITD showing diverse histoscores.

Article Snippet: For the working dilution, the mouse anti-GITRL polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was diluted at 1:150.

Techniques: Expressing, MANN-WHITNEY

Journal:

Article Title: Glucocorticoid-induced tumour necrosis factor receptor-related protein-mediated macrophage stimulation may induce cellular adhesion and cytokine expression in rheumatoid arthritis

doi: 10.1111/j.1365-2249.2007.03363.x

Figure Lengend Snippet: THP-1 cells express both glucocorticoid-induced tumour necrosis factor receptor (TNFR)-related protein (GITR) and GITR ligand (GITRL) and the stimulation of GITR induces intracellular adhesion molecule-1 (ICAM-1) expression. (a) THP-1 cells were stained with anti-GITR and anti-GITRL and the fluorescence profiles (empty lines) were overlayed with background staining (filled area, stained with isotype-matching control antibody). (b) THP-1 cells were stimulated with 10 µg/ml anti-GITR monoclonal antibody or isotype-matching control antibody for 2 days. Expression levels of CD54 (ICAM-1) and CD11a were measured using flow cytometry. Histograms from specific staining (empty lines) and background staining (filled area) are compared. These experiments were repeated more than three times with essentially the same results.

Article Snippet: Monoclonal antibodies and immunohistochemistry Monoclonal antibodies (MoAb) for GITR (clone 621) [ 18 ] and GITRL (clone EB11) were purchased from Immunomics (Ulsan, Korea); endotoxin levels in the anti-GITR/GITRL stock solution (2 mg/ml) were below 20 pg/ml (tested with the QCL-1000 chromogenic Limulus amebolyte lysate test method; Bio-Whittaker, Walkersville, MD, USA); MoAb for CD68 (KP1) and CD3 (F7·2.38), and rabbit polyclonal antibody to von Willebrand factor (vWF) (N1505) from Dako (Glostrup, Denmark); monoclonal antibody to intracellular adhesion molecule-1 (ICAM-1) (BBIG-1), mouse IgG 1 and recombinant human GITRL (rhGITRL) from R&D Systems, Inc. (Minneapolis, MN, USA); and anti-CD11a (HI111) antibody from Becton-Dickinson (Mountain View, CA, USA).

Techniques: Expressing, Staining, Fluorescence, Control, Flow Cytometry

Journal:

Article Title: Glucocorticoid-induced tumour necrosis factor receptor-related protein-mediated macrophage stimulation may induce cellular adhesion and cytokine expression in rheumatoid arthritis

doi: 10.1111/j.1365-2249.2007.03363.x

Figure Lengend Snippet: Stimulation of glucocorticoid-induced tumour necrosis factor receptor (TNFR)-related protein (GITR) induces intracellular adhesion molecule-1 (ICAM-1)-dependent cellular aggregation/adhesion that was enhanced by extracellular matrix proteins (ECM) proteins. (a) carboxyl fluorescein diacetate succinimidyl ester-labelled THP-1 cells were stimulated with 10 µg/ml anti-GITR monoclonal antibody (MoAb) or control antibody for 2 days in the presence or absence of laminin (10 µg/ml) that had been precoated on the culture plate. Cellular aggregates were visualized using fluorescence microscope. (b) THP-1 cells were stimulated with 10 µg/ml anti-GITR MoAb or isotype matching control antibody (mIgG) for 2 days in the presence or absence of collagen, fibronectin and laminin (10 µg/ml) which had been precoated on the culture plate. After the removal of non-adherent cells, the amount of adherent cells were measured as described in Materials and methods. Unstimulated THP-1 cells were also treated in the same way as a negative control. #P < 0·05, ##P < 0·01, ###P < 0·001 when compared with control. *P < 0·05, **P < 0·01, ***P < 0·001 when compared with cells stimulated without ECM proteins. (c) Experiments were performed as in (b) with the addition of anti-ICAM-1 or anti-CD11a MoAb (10 µg/ml). The same amounts of isotype-matching mouse antibodies were added as a control. **P < 0·01, ***P < 0·001 when compared with positive control.

Article Snippet: Monoclonal antibodies and immunohistochemistry Monoclonal antibodies (MoAb) for GITR (clone 621) [ 18 ] and GITRL (clone EB11) were purchased from Immunomics (Ulsan, Korea); endotoxin levels in the anti-GITR/GITRL stock solution (2 mg/ml) were below 20 pg/ml (tested with the QCL-1000 chromogenic Limulus amebolyte lysate test method; Bio-Whittaker, Walkersville, MD, USA); MoAb for CD68 (KP1) and CD3 (F7·2.38), and rabbit polyclonal antibody to von Willebrand factor (vWF) (N1505) from Dako (Glostrup, Denmark); monoclonal antibody to intracellular adhesion molecule-1 (ICAM-1) (BBIG-1), mouse IgG 1 and recombinant human GITRL (rhGITRL) from R&D Systems, Inc. (Minneapolis, MN, USA); and anti-CD11a (HI111) antibody from Becton-Dickinson (Mountain View, CA, USA).

Techniques: Control, Fluorescence, Microscopy, Negative Control, Positive Control