Journal: PLoS ONE

Article Title: Tumour Necrosis Factor-α Regulates Human Eosinophil Apoptosis via Ligation of TNF-Receptor 1 and Balance between NF-κB and AP-1

doi: 10.1371/journal.pone.0090298

Figure Lengend Snippet: The effect of IL-3, IL-5 and GM-CSF neutralization on the antiapoptotic effect of TNF-α in human eosinophils.

Article Snippet: Other reagents were obtained as follows: ammonium pyrrolidinedithiocarbamate (PDTC) and SR 11302 (( E , E , Z , E )-3-Methyl-7-(4-methylphenyl)-9-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2,4,6,8-nonatetraenoic acid) (Tocris Cookson Ltd., Bristol, UK), monoclonal anti-human Fas [CD95] antibody, clone ZB4 (Kamiya Biomedical Co., Tukwila, WA, USA), monoclonal anti-human TNF-α antibody (Genzyme, Cambridge, MA, USA), monoclonal anti-human CD95 Fas antibody, clone CH-11 and IgM isotype control (Immunotech, Marseille, France), monoclonal anti-human GM-CSF monoclonal antibody (Pharmingen, San Diego, CA, USA), IκBα antibody, actin antibody and horseradish-peroxidase conjugated secondary anti-rabbit antibody (Santa-Cruz Biotechnology, Inc., London, UK), IκBα (phospho S32+S36) antibody (Abcam, Cambridge, UK), horseradish peroxidase conjugated goat anti-mouse antibody (Pierce Biotechnology Inc., Rockford, IL, USA), AP-1 and NF-κB consensus oligonucleotides (Promega Corp., Madison, WI, USA), PD98059 (2-(2-Amino-3-methoxyphenyl)-4 H -1-benzopyran-4-one) (Calbiochem Novabiochem, Nottingham, UK), SP600125 (Anthra[1-9- cd ]pyrazol-6(2 H )-one) (Merck Biosciences, Darmstadt, Germany), soluble recombinant human APO-1/Fas ligand (RhFasL) (Alexis Corp., Laüfelfingen, Switzerland) and tanshinone IIA (1,6,6-Trimethyl-6,7,8,9-tetrahydrophenanthro[1,2-b]furan-10,11-dione Dan Shen ketone) (Enzo Life Sciences AG, Lausen, Switzerland).

Techniques: Neutralization

Journal:

Article Title: Allogeneic T cells induce rapid CD34 + cell differentiation into CD11c + CD86 + cells with direct and indirect antigen-presenting function

doi: 10.1182/blood-2005-11-4330

Figure Lengend Snippet: CD34+ cell proliferation stimulated by allogeneic T cells. Freshly isolated CB CD34+ cells were mixed with or without 5 × 104 irradiated autologous T cells (auto-T cells) or HLA-mismatched PB T cells (allo-T cells) at 1:2 S/R ratio, or medium, and cell proliferation was measured by 3H-Thymidine incorporation assay after 6 days of culture. Results are represented as the mean cpm ± SD of 3 separate experiments. (A) CD34+ cell proliferation was greater after coculture with allogeneic T cells than with autologous T cells or with medium alone (P = .02). (B) When anti-HLA class I (anti-class I) or anti-HLA class II (anti-class II) mAbs were added to the culture, only anti-class II reduced proliferation. Results are represented as the mean cpm ± SD of 3 separate experiments. Only the anti-HLA class II antibody reduced the CD34+ cell proliferation significantly (P = .04). (C) Addition of anti-TNFα (anti-TNF) and anti-GM-CSF (anti-GM) mAbs, or anti-TNFα mAb alone (P = .01), reduced proliferation in 3 separate experiments. *P < .05.

Article Snippet: Where indicated, anti-GM-CSF and/or anti-TNFα antibodies (Pharmingen BD, San Diego, CA) were used in cell culture at 350 ng/mL and 10 μg/mL, respectively, to block cytokine activity.

Techniques: Isolation, Irradiation, Thymidine Incorporation Assay

Journal:

Article Title: Allogeneic T cells induce rapid CD34 + cell differentiation into CD11c + CD86 + cells with direct and indirect antigen-presenting function

doi: 10.1182/blood-2005-11-4330

Figure Lengend Snippet: CTLA4-Ig prevents CD34+ cell proliferation upon stimulation with allogeneic T cells. (A) Proliferation of purified CD34+ cells was assessed after 6 days in culture with medium alone or with irradiated allogeneic T cells (irrad T-cells) and CTLA4-IgG4m (CTLA4-Ig), with or without the addition of exogenous cytokines (GM-CSF/TNFα). CTLA4-Ig significantly inhibited CD34+ cell proliferation (P = .05). Results are represented as the mean cpm ± SD of 3 separate experiments. (B) The number of CFU-Cs and CFU-GMs generated from CD34+ cells in clonogenic assays was not affected by addition of CTLA4-Ig. The results are represented as mean colonies ± SD in 3 separate experiments. *P = .05.

Article Snippet: Where indicated, anti-GM-CSF and/or anti-TNFα antibodies (Pharmingen BD, San Diego, CA) were used in cell culture at 350 ng/mL and 10 μg/mL, respectively, to block cytokine activity.

Techniques: Purification, Irradiation, Generated

Journal:

Article Title: Allogeneic T cells induce rapid CD34 + cell differentiation into CD11c + CD86 + cells with direct and indirect antigen-presenting function

doi: 10.1182/blood-2005-11-4330

Figure Lengend Snippet: Differentiation of CD34+ cells upon culture with allogeneic T cells is mediated by endogenous GM-CSF and TNFα. Purified CD34+ cells were cultured with irradiated allogeneic T cells alone (control) or in the presence of anti-GM-CSF (anti-GM-CSF), anti-TNFα (anti-TNFα) mAb, or both (anti-GM-CSF anti-TNFα), then analyzed by flow cytometry for the expression of CD86 and CD11c. Markers were set in each histogram based upon isotype controls in order to calculate the percentage of positive cells.

Article Snippet: Where indicated, anti-GM-CSF and/or anti-TNFα antibodies (Pharmingen BD, San Diego, CA) were used in cell culture at 350 ng/mL and 10 μg/mL, respectively, to block cytokine activity.

Techniques: Purification, Cell Culture, Irradiation, Flow Cytometry, Expressing

Journal:

Article Title: Allogeneic T cells induce rapid CD34 + cell differentiation into CD11c + CD86 + cells with direct and indirect antigen-presenting function

doi: 10.1182/blood-2005-11-4330

Figure Lengend Snippet: Potent allo-antigen (allo-Ag)-presenting function of CD34-derived APCs induced by allogeneic T cells. Purified CD34+ cells, either freshly isolated (□), after culture with irradiated allogeneic T cells for 6 days (Irrad Allo-T cells, ▪), or with irradiated allogeneic T cells and anti-GM-CSF plus anti-TNFα blocking antibodies (blocking MAbs, ), were irradiated and mixed with blood T-cell responders from the same allogeneic donor at a 1:2 ratio in MLC. T-cell alloreactivity was measured by 3H-Thymidine incorporation assay after 6 days of culture. Results are the mean SI ± SD of 4 separate experiments. *P = .05.

Article Snippet: Where indicated, anti-GM-CSF and/or anti-TNFα antibodies (Pharmingen BD, San Diego, CA) were used in cell culture at 350 ng/mL and 10 μg/mL, respectively, to block cytokine activity.

Techniques: Derivative Assay, Purification, Isolation, Irradiation, Blocking Assay, Thymidine Incorporation Assay

Journal: Nature Reviews. Immunology

Article Title: GM-CSF-based treatments in COVID-19: reconciling opposing therapeutic approaches

doi: 10.1038/s41577-020-0357-7

Figure Lengend Snippet: History and current status of GM-CSF-based therapies undergoing assessment in patients with COVID-19

Article Snippet: Roivant , Gimsilumab (anti-GM-CSF) , Phase I HVs + RA Phase I HVs , No announcements , RCT (2 arms, N = 270, NCT04351243).

Techniques: