Macromolecular oxidative alterations have been analysed in vitro in anisotonic suspensions of mouse splenocytes. Both hypertonicity and hypotonicity induced the generation of thiobarbituric acid reactive species (TBARS) and carbonylation of the proteins, which took place along with cell death. Addition of antioxidants partially inhibited oxidative changes in isotonic and hypotonic suspensions. Anisotonic shock of mouse splenocytes proved to be an inducer of oxidative stress. The oxidative macromolecular alterations might contribute to pathogenesis of cell death caused by osmotic stress.