Human immunodeficiency virus type 1 (HIV-1) has the ability to adapt to the host environment by escaping from host immune responses. We previously observed that escape from humoral immunity, both at the individual and at a population level, coincided with longer variable loops and an increased number of potential N-linked glycosylation sites (PNGS) in the viral envelope glycoprotein (Env) and, in particular, in variable regions 1 and 2 (V1V2). Here, we provide several lines of evidence for the role of V1V2 in the resistance of HIV-1 to neutralizing antibodies. First, we determined that the increasing neutralization resistance of a reference panel of tier-categorized neutralization-sensitive and -resistant HIV-1 variants coincided with a longer V1V2 loop containing more PNGS. Second, an exchange of the different variable regions of Env from a neutralization-sensitive HIV-1 variant into a neutralization-resistant escape variant from the same individual revealed that the V1V2 loop is a strong determinant for sensitivity to autologous-serum neutralization. Third, exchange of the V1V2 loop of neutralization-sensitive HIV-1 variants from historical seroconverters with the V1V2 loop of neutralization-resistant HIV-1 variants from contemporary seroconverters decreased the neutralization sensitivity to CD4-binding site-directed antibodies. Overall, we demonstrate that an increase in the length of the V1V2 loop and/or the number of PNGS in that same region of the HIV-1 envelope glycoprotein is directly involved in the protection of HIV-1 against HIV-specific neutralizing antibodies, possibly by shielding underlying epitopes in the envelope glycoprotein from antibody recognition.The HIV-1 envelope glycoprotein (Env) is a major target of the humoral immune response in HIV-1-infected individuals. Antibodies directed against Env can be detected early in infection and are able to neutralize autologous virus variants with increasing titers over time in most patients (1,35,40,51). HIV-1 Env has developed multiple mechanisms to evade neutralizing antibodies, including the inaccessibility of relevant epitopes due to the trimeric structure of Env, the density of glycosylation, and the presence of occluding variable loops on the outer domain of Env (11,14,17,22,51). Moreover, some epitopes for neutralizing antibodies only emerge after the conformational changes that occur upon the engagement of Env with the CD4 receptor, when spatial constraints between cell and viral membrane no longer allow binding of the relatively large immunoglobulins to Env (21, 22, 23, 29, 52).The HIV-1 Env is synthesized as a gp160 precursor protein, which is subsequently cleaved into two subunits, surface protein gp120 and transmembrane protein gp41. Three subunits of gp120 bind noncovalently to three subunits of gp41 to form a trimeric complex on the surface of the virion. Gp120 is composed of five conserved regions (C1 to C5) that are interspersed with 5 variable regions (V1 to V5) (47). The conserved regions form a central core consisting of an inner dom...