mouse monoclonal anti cd163 ab  (Novus Biologicals)

Journal: Reproduction (Cambridge, England)

Article Title: MEHP-Induced Rat Testicular Inflammation Does Not Exacerbate Germ Cell Apoptosis

doi: 10.1530/REP-18-0093

Figure Lengend Snippet: Expression of CD11bc, CD68 and CD163 by interstitial cells was measured 12, 24, and 48 hrs after MEHP treatment (0.7 g/kg, p.o.) or corn oil control. Flow plots indicating staining for (A) CD68 and CD11bc and (B) CD68 and CD163 of CD11bc-positive cells after MEHP treatment or control. Gates indicate the percentage of cells that are double-positive, single-positive, or unstained. Flow plots are representative of n ≥ 3 for each treatment. (C) Bar chart indicating the total number of CD11bc-expressing cells that are negative, single-positive, or double-positive for CD68 and CD163 expression. (D) Bar chart indicating the geometric mean fluorescence intensity (geo MFI) of CD163 cell surface expression of CD11bc+/CD68+/CD163+ cells. (C-D) Statistical analysis was performed by comparing each group of cells with those of control rats for each time point using the unpaired t-test and corrected for multiple comparisons using the Holm-Sidak method. (E) Immunofluorescent detection of cells labelled with anti-CD68 or anti-CD163 antibody in testicular cross sections 12hrs after exposure to MEHP. Images show immunofluorescence overlaid on differential interference contrast (DIC) images and are representative of n ≥ 3 for each treatment. Scale bars = 50μm. (F) Quantification of CD68-positive and CD163-positive cells from immunofluorescent images. Individual data points are shown and means ± S.E.M for n≥3. Statistical analysis was performed using one-way ANOVA and Tukey’s multiple comparisons test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: Sections were incubated with mouse anti-mononuclear phagocyte monoclonal Ab (1:200 dilution; clone IC7; BD Biosciences, San Jose, CA, USA), or mouse anti-rat CD68 (1:200 dilution, MCA341R, Bio-Rad), or mouse anti-CD163 monoclonal Ab (1:200 dilution; clone ED2; Thermo Fischer Scientific) for 1hr at room temperature.

Techniques: Expressing, Staining, Fluorescence, Immunofluorescence

Journal: Reproduction (Cambridge, England)

Article Title: MEHP-Induced Rat Testicular Inflammation Does Not Exacerbate Germ Cell Apoptosis

doi: 10.1530/REP-18-0093

Figure Lengend Snippet: mRNA expression by (A-B) macrophages from animals 12 hours after exposure to MEHP or control or (C-E) testicular macrophages exposed to 200μM MEHP in vitro for 3, 6, or 12 hrs of (A,C) pro-inflammatory, (B,D) anti-inflammatory genes, or (E) Cd163, is presented as fold increase compared to gene expression of control macrophages. Data are means ± S.E.M. for n≥4. Statistical analysis was performed by the (A-B) unpaired two-tailed t-test and (C-E) the unpaired t-test and corrected for multiple comparisons using the Holm-Sidak method. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Sections were incubated with mouse anti-mononuclear phagocyte monoclonal Ab (1:200 dilution; clone IC7; BD Biosciences, San Jose, CA, USA), or mouse anti-rat CD68 (1:200 dilution, MCA341R, Bio-Rad), or mouse anti-CD163 monoclonal Ab (1:200 dilution; clone ED2; Thermo Fischer Scientific) for 1hr at room temperature.

Techniques: Expressing, In Vitro, Two Tailed Test

Journal: Reproduction (Cambridge, England)

Article Title: MEHP-Induced Rat Testicular Inflammation Does Not Exacerbate Germ Cell Apoptosis

doi: 10.1530/REP-18-0093

Figure Lengend Snippet: Before MEHP-exposure of PND28 rats, monocytes were depleted from the circulation using clodronate liposomes as described in materials and methods. (A) Quantification of flow cytometric counts of CD68+ macrophages in the blood. (B) Bar chart indicating the total number of CD11bc-expressing cells that are negative, single-positive, or double-positive for CD68 and CD163 expression. (C) Representative TUNEL images and d) quantification of untreated rats, and rats treated with MEHP that had been pretreated with clodronate liposomes to deplete monocytes from circulation, or control PBS liposomes. Scale bars = 100μm. (D) Bar chart indicating the geometric mean fluorescence intensity (geo MFI) of CD163 cell surface expression of CD11bc+/CD68+/CD163+ cells. Data are means ± S.E.M. for n=3. Statistical analysis was performed using (A,D) one-way ANOVA and (B) two-way ANOVA and Tukey’s multiple comparisons test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: Sections were incubated with mouse anti-mononuclear phagocyte monoclonal Ab (1:200 dilution; clone IC7; BD Biosciences, San Jose, CA, USA), or mouse anti-rat CD68 (1:200 dilution, MCA341R, Bio-Rad), or mouse anti-CD163 monoclonal Ab (1:200 dilution; clone ED2; Thermo Fischer Scientific) for 1hr at room temperature.

Techniques: Expressing, TUNEL Assay, Fluorescence

Journal: Reproduction (Cambridge, England)

Article Title: MEHP-Induced Rat Testicular Inflammation Does Not Exacerbate Germ Cell Apoptosis

doi: 10.1530/REP-18-0093

Figure Lengend Snippet: Before MEHP-treatment of PND28 rats, monocytes and/or neutrophils were depleted from the circulation by two i.v. injections of an anti-PMN Ab as described in materials and methods. (A) Quantification of flow cytometric counts of neutrophils and CD68+ macrophages in the blood. (B) Bar chart indicating the total number of CD11bc-expressing cells that are negative, single-positive, or double-positive for CD68 and CD163 expression. (C) Representative images and (D) summarized data of myeloperoxidase staining. Scale bars = 100μm. (E) Representative TUNEL images and (F) quantification of TUNEL-positive tubules of rats treated with MEHP with or without neutrophil depletion from the circulation. (G) Bar chart indicating the geometric mean fluorescence intensity (geo MFI) of CD163 cell surface expression of CD11bc+/CD68+/CD163+ cells. Data are means ± S.E.M. for n= 4-7. Statistical analysis was performed using two-way ANOVA and Tukey’s multiple comparisons test. **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: Sections were incubated with mouse anti-mononuclear phagocyte monoclonal Ab (1:200 dilution; clone IC7; BD Biosciences, San Jose, CA, USA), or mouse anti-rat CD68 (1:200 dilution, MCA341R, Bio-Rad), or mouse anti-CD163 monoclonal Ab (1:200 dilution; clone ED2; Thermo Fischer Scientific) for 1hr at room temperature.

Techniques: Expressing, Staining, TUNEL Assay, Fluorescence

Journal: Arthritis Research & Therapy

Article Title: Circulating levels of soluble MER in lupus reflect M2c activation of monocytes/macrophages, autoantibody specificities and disease activity

doi: 10.1186/ar4407

Figure Lengend Snippet: Soluble Mer is released by M2c monocytes/macrophages and correlates with circulating levels of soluble CD163 in systemic lupus erythematosus, and soluble CD163, like soluble Mer, correlates with Systemic Lupus Erythematosus Disease Activity Index score. Levels of soluble Mer (sMer) (A) and soluble Axl (sAxl) (B) were measured in supernatants of monocytes/macrophages cultured for 3 days in the presence of medium alone (resting M0), interferon γ (IFN-γ), granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin 17 (IL-17) (classically activated and/or proinflammatory M1 and M17), IL-4, transforming growth factor β (TGF-β), IL-10, macrophage colony-stimulating factor (M-CSF), dexamethasone (Dex), alone or in combination (alternatively activated and/or regulatory M2a and M2c cells). In 40 consecutive patients in our cohort, potential correlations between plasma levels of sMer (C) or sAxl (D) and soluble CD163 (sCD163) were examined. Levels of sCD163 were also analyzed according to circulating autoantibodies (E) and disease activity (F) . αchrom., antichromatin; SLEDAI, Systemic Lupus Erythematosus Disease Activity Index. * P < 0.05; ** P < 0.01; n.s., not significant.

Article Snippet: For sCD163 quantitation in supernatants, we used a mouse monoclonal anti-CD163 Ab (clone EDHu-1; Novus Biologicals) for capture, recombinant human CD163 (R&D Systems) for the standard curve and a biotinylated goat polyclonal anti-CD163 Ab (R&D Systems) for detection.

Techniques: Activity Assay, Cell Culture, Clinical Proteomics

Journal: Arthritis Research & Therapy

Article Title: Circulating levels of soluble MER in lupus reflect M2c activation of monocytes/macrophages, autoantibody specificities and disease activity

doi: 10.1186/ar4407

Figure Lengend Snippet: Interferon α/β is required for production of soluble Axl, and combining M2c polarizing conditions with type I interferon exposure inhibits soluble Mer production and enhances soluble Axl release. Levels of soluble Mer (sMer) (A) , soluble CD163 (sCD163) (B) and soluble Axl (sAxl) (C) were measured in supernatants of monocytes/macrophages cultured for 3 days in the presence or absence of type I interferons (IFN-α or IFN-β), macrophage growth factors (macrophage colony-stimulating factor (M-CSF) or granulocyte macrophage colony-stimulating factor (GM-CSF)), M2c polarizing agents (interleukin 10 (IL-10) or dexamethasone) or combinations of these. On day 2, low doses of lipopolysaccharide (LPS) were added for an additional 24 hours to stimulate ectodomain shedding of membrane receptors. Surface expression levels of Mer receptor tyrosine kinase (MerTK), Axl and CD163 were measured by flow cytometry (D) . Numbers shown in (D) refer to mean fluorescence intensity values. Data are representative of three independent experiments. Dex, dexamethasone. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: For sCD163 quantitation in supernatants, we used a mouse monoclonal anti-CD163 Ab (clone EDHu-1; Novus Biologicals) for capture, recombinant human CD163 (R&D Systems) for the standard curve and a biotinylated goat polyclonal anti-CD163 Ab (R&D Systems) for detection.

Techniques: Cell Culture, Membrane, Expressing, Flow Cytometry, Fluorescence