LA loaded moDCs induce upregulated expressions of TLR4 and TLR2 on human γδ T cells and the response of LA-activated γδ T cells to LA is closely associated with TLR4. (A) Tested cells were stained with FITC-conjugated anti-TCRγδ mAb and PE-conjugated anti-TLR4/TLR2 mAbs, and then determined by flow cytometry. The resting purified γδ T cells did not express TLR4 or TLR2 (right panel). After 7 days' co-culture with LA-loaded moDCs, TLR2+ TCRγδ+ and TLR4+ TCRγδ+ T cells accounted for 19% and 40% of tested cells, respectively (left panel). Up-panel is isotype antibody control. This result is shown as a representative experiment of three independent ones. (B) LA-activated γδ T cells were incubated with mAb TLR2, TLR4 or TLR2 plus TLR4 and rested for 24 h in medium. Autologous moDCs were preincubated with mAbs against CD1b, CD1c CD1b plus CD1c mAb at 37°C for 2 h. Then, they were thoroughly washed respectively, co-cultured with LA for 48 hour. During the last 8 h, 1 μCi of 3H-TdR per-well was added. 3H-TdR incorporation was measured. The results are expressed as mean cpm ± SD of five independent experiments. *P < 0.05, **P > 0.05, when compared with the group without mAb.