Fig. 3.

Inhibition of cytokine/CD40L gene expression by Ly294002. (a) CD4⁺ peripheral blood T lymphocytes (PBT) and CD4⁺ LPT were cultured in the presence or absence of CD2 monoclonal antibody (mAb) M1 (1 μg/ml), M2 (1 μg/ml) and 3PT (0·33 μg/ml) for 4 h. Ly294002 was added 30 min prior to stimulation. Interleukin (IL)-2, interferon (IFN)-γ and granulocyte–macrophage colony-stimulating factor (GM-CSF) mRNA expression was determined by quantitative reverse transcribed polymerase chain reaction. Relative inhibition (in percentage) of cytokine/CD40L gene expression by Ly294002 is shown. Data are expressed as mean ± standard deviation (n = 3–6). The effectiveness of cytokine/CD40L inhibition was tested statistically for 1 μM Ly294002 (n = 6). (b) CD2-induced AKT phosphorylation in CD4⁺ PBT and lamina propria T lymphocytes (LPT) in the presence or absence of Ly294002 was quantified by densitometry (n = 3–4).