The effect of pre-strain on tensile behaviour of 316L austenitic stainless steel was investigated, focusing on strain rate sensitivity, temperature sensitivity and strain hardening. Experimental data showed that strain rate sensitivity, temperature sensitivity and strain hardening were weakening with pre-strain. Meanwhile, the variation of microstructure with pre-strain was observed by optical microscopy, scanning electronic microscopy and X-ray diffraction. Then, the mechanical properties of pre-strained material were correlated with the increase in dislocation density and mechanical twinning with pre-strain. Finally, an improved Arrhenius-PS model considering the effect of pre-strain was developed. This paper is part of a thematic issue on Nuclear Materials.