Journal: Mediators of Inflammation

Article Title: IL-1 β and TNF α Promote Monocyte Viability through the Induction of GM-CSF Expression by Rheumatoid Arthritis Synovial Fibroblasts

doi: 10.1155/2014/241840

Figure Lengend Snippet: Effect of IL-1 β and TNF α on M-CSF, IL-34, and GM-CSF expression in RA SF . SF were stimulated by IL-1 β (25 ng/mL) or TNF α (25 ng/mL) for 24 hours. (a)–(c) RNA was harvested and a qPCR for M-CSF, IL-34, and GM-CSF expression was performed. Results are given as the relative expression using hHPRT gene as housekeeping gene. The bars represent the mean and each plots one biological replicate ( n = 6 patients). (d) GM-CSF concentration (pg/mL) was assessed by ELISA. Results are given as the mean (± SEM). * P < 0.05; ** P < 0.001; *** P < 0.0001.

Article Snippet: GM-CSF levels were measured in RA SF conditioned media by ELISA assay (Human GM-CSF DuoSet ELISA development kit, R&D Systems).

Techniques: Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Infection and Immunity

Article Title: Granulocyte-Macrophage Colony-Stimulating Factor- and Tumor Necrosis Factor Alpha-Mediated Matrix Metalloproteinase Production by Human Osteoblasts and Monocytes after Infection with Brucella abortus

doi: 10.1128/iai.00934-10

Figure Lengend Snippet: FIG. 1. (A) Infection and replication of B. abortus within human osteoblasts (hFOB cell line). After infection at different MOIs, cells were incubated with antibiotics to kill extracellular bacteria. Cell ly- sates obtained at different times postinfection (p.i.) were plated on agar to determine intracellular CFU. (B and C) MMP-2 production at 48 h p.i. by B. abortus-infected osteoblasts as determined by ELISA (B) and zymography (C). (D) GM-CSF production by B. abortus- infected osteoblasts at 48 h p.i. (E) Inhibition of MMP-2 secretion by B. abortus-infected osteoblasts in the presence of an anti-GM-CSF neutralizing antibody or an isotype control. The data shown are from a representative experiment of five performed. *, P 0.05; **, P 0.01; ***, P 0.001 versus control.

Article Snippet: Neutralization experiments were performed with an antibody against the human TNF receptor I (anti-TNFRI, clone 16805) or its isotype control (both from R&D Systems), an anti-TNFneutralizing antibody (clone MAb1) or its isotype control (both from BD Biosciences, San Diego, CA), an anti-GM-CSF neutralizing antibody (clone BVD223B6) or its isotype control (BD Biosciences), or the natural antagonist IL-1Ra (R&D Systems).

Techniques: Infection, Incubation, Bacteria, Enzyme-linked Immunosorbent Assay, Zymography, Inhibition, Control

Journal: Infection and Immunity

Article Title: Granulocyte-Macrophage Colony-Stimulating Factor- and Tumor Necrosis Factor Alpha-Mediated Matrix Metalloproteinase Production by Human Osteoblasts and Monocytes after Infection with Brucella abortus

doi: 10.1128/iai.00934-10

Figure Lengend Snippet: FIG. 7. MMP-9 production by monocytes stimulated with superna- tants from B. abortus-infected osteoblasts (added at a 1/2, 1/5, or 1/10 proportion) or from uninfected osteoblasts, as determined by zymo- graphy (A). Inhibition of the stimulating effect by pretreatment of supernatants with a neutralizing antibody to GM-CSF or an isotype control for 1 h before addition to monocytes determined by ELISA (B) and zymography (C). *, P 0.05 versus untreated supernatants.

Article Snippet: Neutralization experiments were performed with an antibody against the human TNF receptor I (anti-TNFRI, clone 16805) or its isotype control (both from R&D Systems), an anti-TNFneutralizing antibody (clone MAb1) or its isotype control (both from BD Biosciences, San Diego, CA), an anti-GM-CSF neutralizing antibody (clone BVD223B6) or its isotype control (BD Biosciences), or the natural antagonist IL-1Ra (R&D Systems).

Techniques: Infection, Inhibition, Control, Enzyme-linked Immunosorbent Assay, Zymography

Journal: Infection and Immunity

Article Title: Granulocyte-Macrophage Colony-Stimulating Factor- and Tumor Necrosis Factor Alpha-Mediated Matrix Metalloproteinase Production by Human Osteoblasts and Monocytes after Infection with Brucella abortus

doi: 10.1128/iai.00934-10

Figure Lengend Snippet: FIG. 8. GM-CSF produced by B. abortus-infected osteoblasts induces TNF- and IL-1 production in monocytes. TNF- (A) and IL-1 (B) production by monocytes stimulated with supernatants from B. abortus-infected osteoblasts. This stimulating effect is inhibited by pretreatment of supernatants with a neutralizing antibody to GM-CSF for 1 h before addition to monocytes (C and D, respectively).

Article Snippet: Neutralization experiments were performed with an antibody against the human TNF receptor I (anti-TNFRI, clone 16805) or its isotype control (both from R&D Systems), an anti-TNFneutralizing antibody (clone MAb1) or its isotype control (both from BD Biosciences, San Diego, CA), an anti-GM-CSF neutralizing antibody (clone BVD223B6) or its isotype control (BD Biosciences), or the natural antagonist IL-1Ra (R&D Systems).

Techniques: Produced, Infection

Journal: Infection and Immunity

Article Title: Granulocyte-Macrophage Colony-Stimulating Factor- and Tumor Necrosis Factor Alpha-Mediated Matrix Metalloproteinase Production by Human Osteoblasts and Monocytes after Infection with Brucella abortus

doi: 10.1128/iai.00934-10

Figure Lengend Snippet: FIG. 10. Proposed model for the production of MMPs by osteo- blasts and monocytes during Brucella infection. Osteoblasts would produce MMP-2 not only in response to infection but also in response to TNF- produced by adjacent infected monocytes. MMP-2 produc- tion by infected osteoblasts would depend on the autocrine action of GM-CSF. Monocytes would produce MMP-9 in response to infection and also in response to GM-CSF produced by adjacent infected os- teoblasts. The later effect would be mediated by the autocrine action of TNF- produced by monocytes in response to GM-CSF.

Article Snippet: Neutralization experiments were performed with an antibody against the human TNF receptor I (anti-TNFRI, clone 16805) or its isotype control (both from R&D Systems), an anti-TNFneutralizing antibody (clone MAb1) or its isotype control (both from BD Biosciences, San Diego, CA), an anti-GM-CSF neutralizing antibody (clone BVD223B6) or its isotype control (BD Biosciences), or the natural antagonist IL-1Ra (R&D Systems).

Techniques: Infection, Produced