The shear properties of the permafrost-structure interface play a crucial role in analyzing the performance of engineering structures in cold regions. Direct shear tests were carried out on the permafrost-concrete structural interface by means of self-improved and optimized temperature-controlled direct shear apparatus to investigate the effects of temperature, moisture content and freeze-thaw cycles. Test results were used to establish a shear characteristic model under coupled influence of temperature and humidity. Preparation of specimens consisting of precast C30 concrete blocks and soil samples taken from borehole cores of the pile foundation of the Middle Bridge No. 3 on the road from Sai Township to Qiuluo Village, Sakya County, Tibet, China, and were treated to different effective confining pressure (σn = 300 kPa, 400 kPa, 500 kPa and 600 kPa), different temperatures (T = −20 °C, −15 °C, −5 °C, 0 °C, 5 °C, 15 °C, and 20 °C), different moisture contents (ω = 5 %, 10 %, 15 %, and 20 %), and different numbers of freeze-thaw cycles (N = 0, 1, 3, 5, 7and11) from −15 °C to 15 °C, test results are given and analyzed. The peak shear stress (τp) is significantly higher than the residual stress (τr) at negative temperature, τp is greater than 1.5 times to 3.2 times of τr. τp is greatly affected by temperature changes, τp under negative temperature is 17 % higher than that under positive temperature, because the water in the soil freezes at negative temperatures, forming a concrete-ice-soil bond interface, which increases the peak strength of the interface. While τr is insensitive to temperature changes. At negative temperatures, the cohesion (c') changes less with increasing water content (ω), and the effect of ω on the external friction angle (φ′) is more significant. At positive temperature, the cohesion decreases with increasing ω, and the effect of ω on the external friction angle is not significant. When N < 5, the cohesion increases with the increase of N; when N = 5, c' reaches the maximum value; when N > 5, c' decreases with the increase of N. Moreover, regardless of the type of shear behavior, the first five freeze-thaw cycles have the greatest impact on the external friction angle. Compared with the sample that has not experienced freeze-thaw cycles, when N = 5, the total decrease in the external friction angle of the sample is 25.6 %.