Ultrasonic fatigue tests have been performed in austenitic stainless steel, SUS316NG, in order to investigate giga-cycle fatigue strength of pre-strained materials, i.e. 5, 10 and 20% tensile pre-strains and -20% compressive pre-strain. The pre-strains were applied before specimen machining. The austenitic stainless steels are known to exhibit remarkable self-heating during the fatigue experiment. Therefore, heat radiation method was established by setting fatigue specimens in a low temperature chamber at about -100°C. The self-heating was controlled by intermittent loading condition, which enabled us to maintain the test section of the specimens at about room temperature. The results revealed that the fatigue strength increased with increasing pre-strain levels. Fish-eye fracture was observed for -20% pre-strained specimen fractured at 4.11×10⁷ cycles, while the other specimens exhibited ordinary fatigue fracture surface originated from stage I facet on the specimen surface. The increase in fatigue limit was predicted by Vickers hardness, HV, which depended on the size of indented region. The prediction was successful using HV values obtained by the size of the indented region similar to those of the stage I facets.