Transduced αT2ib inhibits lipopeptide-but not LPS-induced macrophage activation. TLR2 specific induction of cytokine mRNA accumulation and protein production. RAW264.7 macrophages were challenged with 100 ng/ml Pam3CSK4 or 100 ng/ml LPS for 4 h at room temperature. A, Cytokine mRNA accumulation in RAW264.7 macrophages was analysed upon RNA isolation and cDNA synthesis. Specific PCR amplificates were analysed semi-quantitatively (HPRT, hypoxanthine guanine phophoribosyl transferase). B, RAW264.7 macrophages were fixed/permeabilized and the TNFα release inhibited during stimulation. Intracellular cytokine was analysed by flow cytometry (black areas, specific staining; white areas, unspecific staining with isotype control mAb). All viable infected GFP+ cells were analysed. C, Analysis of cytokine release to cell supernatant by ELISA (white columns, non infected; grey columns, AdVGFP infected; black columns, AdVαT2ib infected). Results of four independent experiments (p < 0.001).