Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Differential expression of adenosine A3 receptors controls adenosine A2A receptor-mediated inhibition of TLR responses in microglia.

doi: 10.4049/jimmunol.0803383

Figure Lengend Snippet: FIGURE 4. The enhanced inhibitory effect of NECA is mediated by the enhanced contribution of A2AR-mediated signaling and the decreased contribution of A3R-mediated signaling. Unstimulated and activated microglia were exposed to 100 ng LPS/ml in the presence or absence of 10 M NECA in combination with A1, A2A, A2B, and A3 receptor antagonists. After 16 h, TNF- (A) and IL-12p40/p70 (B) levels were measured and expressed as relative values of exposure to 100 ng LPS/ml alone. DPCPX was used as an A1R, SCH58261 as an A2AR, alloxazine as an A2BR and VUF5574 as an A3R antagonist. All antagonists were used at 10 M concentrations. Solid lines in the graph indicate relative cytokine production levels of exposure to 100 ng LPS/ml 10 M NECA without antagonists of unstimulated microglia, the dashed lines indicate activated microglia. , p 0.05; Student’s t test. TNF- (C) and IL-12p40/p70 (D) production after exposure to 100 ng LPS/ml in the presence or absence of 10 M CGS21680, an A2AR-selective agonist, and/or of 10 M VUF5574, an A3R antagonist. Cytokine production is expressed relative to production after exposure to 100 ng LPS/ml alone for unstimulated and activated microglia. , p 0.05; Student’s t test. TNF- (E) and IL-12p40/p70 (F) production after exposure to 100 ng LPS/ml different concentrations of CGS21680, in the presence or absence of A3R antagonist, is expressed relative to production after exposure to 100 ng LPS/ml alone for unstimulated and activated microglia. All data are representative for three independent experiments using different donors.

Article Snippet: Cells were washed and incubated for 30 min with anti-adenosine A3 receptor Abs (SP055P, Acris Antibodies) or total rabbit IgG (AbD Serotec) as an isotype control.

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