Recently, BST-2 has been identified as an effective cellular factor that prevents the release of human immunodeficiency virus type 1 and other enveloped viruses by tethering virus particles to the cell surface. Here, we showed that the production of HIV-1 virus-like particles was markedly inhibited by BST-2. Both the transient and stable expressing of BST-2 had the same function and Vpu rescued the release of HIV-1 VLP in the presence of human BST-2. Consistent with a direct tethering mechanism, we confirmed that proteolysis releases restricted virions and further showed that this removed the ectodomain of BST-2 from the cell surface.