Journal: Purinergic Signalling

Article Title: Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4 + T lymphocytes activation

doi: 10.1007/s11302-005-8077-9

Figure Lengend Snippet: ATPγS inhibits IFN-γ secretion in primary human CD4 + T cells following activation mediated by anti-CD3 plus anti-CD28 but not by PMA plus anti-CD28. The cells were incubated for 24 h either with PMA (1 ng/ml) and two concentrations of soluble anti-CD28 (1 or 5 µg/ml) (A) or with a combination of pre-coated anti-CD3 and soluble anti-CD28 (1 µg/ml) (B), in the continuous presence or absence of ATPγS, that was added at the same time as PMA/CD28 or anti-CD3/anti-CD28. In the absence of stimulation, no IFN-γ was detectable in the supernatant (data not shown). Data represent the mean ± S.D. of triplicate experimental points obtained in one representative experiment of three.

Article Snippet: The anti- CD3 monoclonal antibody (mAb) OKT3 (Orthoclone OKT3®) was provided by Janssen-Cilag (Berchem, Belgium) and the anti-CD28 mAb (clone CD28.2) by BD Pharmingen (San Diego, CA, USA).

Techniques: Activation Assay, Incubation

Journal: Purinergic Signalling

Article Title: Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4 + T lymphocytes activation

doi: 10.1007/s11302-005-8077-9

Figure Lengend Snippet: ATPγS has no effect on the calcium response induced by TCR stimulation with anti-CD3 and anti-CD28. Cells were loaded with FURA 2-AM, and intracellular calcium mobilization was followed on a spectrofluorometer (LS50B, Perkin Elmer). CD4 + T lymphocytes were activated by cross-linking the anti-CD3/anti-CD28 mAb with goat anti-IgG (▾) without (A) or with 100 µM ATPγS added (▿) to the cells 2 min before cross-linking (B). Data are from one representative experiment out of three.

Article Snippet: The anti- CD3 monoclonal antibody (mAb) OKT3 (Orthoclone OKT3®) was provided by Janssen-Cilag (Berchem, Belgium) and the anti-CD28 mAb (clone CD28.2) by BD Pharmingen (San Diego, CA, USA).

Techniques:

Journal: Purinergic Signalling

Article Title: Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4 + T lymphocytes activation

doi: 10.1007/s11302-005-8077-9

Figure Lengend Snippet: (A) ATPγS pre-treatment inhibits the phosphorylation of ERK1/2 (a) and p-38 (b) induced by TCR stimulation with anti-CD3 and anti-CD28 mAb. ATPγS 100 µM was added to the cells 10 min before activation with antibodies. The same amount of protein for each condition was electrophoresed on a 12% SDS-polyacrylamide gel. (B) ATPγS pretreatment inhibits the phosphorylation of PKB induced by TCR stimulation with anti-CD3 and anti-CD28 mAb. ATPγS 100 µM was added to the cells 10 min before activation with antibodies. The same amount of protein (20 µg) for each condition was electrophoresed on a 12% SDS-polyacrylamide gel.

Article Snippet: The anti- CD3 monoclonal antibody (mAb) OKT3 (Orthoclone OKT3®) was provided by Janssen-Cilag (Berchem, Belgium) and the anti-CD28 mAb (clone CD28.2) by BD Pharmingen (San Diego, CA, USA).

Techniques: Activation Assay

Journal: Purinergic Signalling

Article Title: Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4 + T lymphocytes activation

doi: 10.1007/s11302-005-8077-9

Figure Lengend Snippet: Effect of ATPγ s , ATP and ADP on IFN-γ secretion following CD3 and CD28 stimulation.

Article Snippet: The anti- CD3 monoclonal antibody (mAb) OKT3 (Orthoclone OKT3®) was provided by Janssen-Cilag (Berchem, Belgium) and the anti-CD28 mAb (clone CD28.2) by BD Pharmingen (San Diego, CA, USA).

Techniques: Inhibition