Objective To investigate the role of interleukin (IL)-12 in regulation of the expression of L-selectin (CD62L) on γδT cells. Methods After induction and proliferation, human peripheral activated γδT cells were primarily and secondarily stimulated with Mtb-Ag. Cells were cultured with different cytokines (IFN-γ, IL-12+anti-IFN-γ, IL-12+anti-IL-12 and IL-12+IFN-γ) and different concentrations of IL-12 (0 ng/mL, 0.1 ng/mL and 1 ng/mL), respectively. Cell cultured for 0 d, 3 d, 5 d and 7 d were collected, and the expression of CD62L on γδT cells was detected by flow cytometry. Results After primary and secondary stimulation with Mtb-Ag, the expression of CD62L on γδT cells in IL-12 group, IL-12+anti-IFN-γ group and IL-12+ IFN-g group was significantly higher than that in blank control group (P<0.05), while the expression of CD62L on γδT cells in IFN-γ group and IL-12+anti-IL-12 group was not significantly different from that in blank control group (P>0.05). With the increase of concentrations of IL-12, the expression of CD62L on γδT cells increases. On the third day and fifth day after primary stimulation with Mtb-Ag, the expression of CD62L in 0.1 ng/mL IL-12 group and 1.0 ng/mL IL-12 group was significantly higher than that in negative control group (P<0.05), and the expression of CD62L in 1.0 ng/mL IL-12 group was significantly higher than that in 0.1 ng/mL IL-12 group (P<0.05). The expression of CD62L decreased after secondary stimulation with Mtb-Ag, while the decrease of expression of CD62L slowed down after treatment with IL-12, and the effect was more significant for 1.0 ng/mL IL-12 compared with 0.1 ng/mL IL-12 (P<0.05). Conclusion The expression of CD62L is dose-dependently regulated by IL-12 when γδT cells are activated.