The occurrence of an interactive effect between warp and weft of woven fabrics under practical working conditions is an important concern for researchers. A novel 3D loading device has therefore been developed to elucidate information on materials subjected to loadings in multiple dimensions. This paper investigated the interactive effects between warp and weft in biaxial tension. Strain fields were characterized by digital speckle correlation method and Poisson's ratio was calculated at 0°in the warp direction. The effects of different cutting angles and loading threshold values at 0°on the interactive effect were also evaluated. An interactive effect between warp and weft existed, and the influence of load at weft to warp was larger than warp to weft. Weft produced larger strain fields in both directions. As the load in the warp direction linearly increased, force values nonlinearly increased while load ratio decreased. Poisson's ratio, however, was practically unchanged. Loading mean value curves at x and y axes had sine curve characteristics. The greatest interactive effect occurred when the warp load was 65.3% of the biaxial breaking strength.