Abstract: Previous studies showed that heat-shock protein 60 (HSP60) was known to function as a molecular chaperone and is an important factor in the innate immune system in mammals. However, little was known about the physiological relevance of HSP60 in marine invertebrates. This study focuses on long-term monitoring of the differential expression of LvHSP60 in shrimp Litopenaeus vannamei in response to environmental stress. The thermal aggregation assay elucidated that LvHSP60 was an effective chaperone. It also suggested that LvHSP60 may employ the cell’s intrinsic mechanism to start the immunizing process. Using quantitative real-time PCR to monitor gene expression showed that LvHSP60 was variable under different stresses including environmental stress and pathogenic infection. LvHSP60 was speculated to regulate the adaptive responses to overcome environmental stresses. In conclusion, our study proved that LvHSP60 plays an important role in the intrinsic immune system and stress responses of shrimp.