NK cells play critical roles in the first line of defense against viruses and other pathogens. However, the factors that control NK cell recruitment into local sites to exert effector functions during viral infection remain poorly understood. In this study, we found that murine NK cells in various organs could be divided into CD62L− and CD62L+ subsets, the latter of which were less abundant in the liver and exhibited a relatively mature NK cell phenotype and a stronger cytotoxic function. Moreover, NK cells acquired CD62L expression after birth, and the frequency of CD62L+ NK cells gradually increased during postnatal development. In models of polyinosinic-polycytidylic acid administration and adenovirus infection in vivo, CD62L+ NK cell frequency and absolute numbers in the liver rapidly and markedly increased as a result of the augmented differentiation of CD62L− to CD62L+ NK cells and recruitment of peripheral mature NK cells to the liver. However, blocking CD62L prior to administering viral stimuli in vivo abolished viral stimulation–induced NK cell accumulation and maturation in the liver. Collectively, these data suggest that CD62L marks a mature NK cell subset, as well as affects the magnitude of the local NK cell response to viral infection.