Human immunodeficiency virus (HIV)-2 differs from HIV-1 in its relative lower transmissibility and pathogenicity. To understand the virologic basis of these differences, the nef gene from HIV-2-seropositive persons was analyzed because of its importance for disease progression in the genetically related simian immunodeficiency virus (SIV[MAC]). Proviral nef sequences from 60 HIV-2-infected persons were amplified from peripheral blood lymphocytes, and nef open-reading frames were screened by a transcription and translation assay for the presence of full-length (32- to 36-kDa) or truncated (<32 kDa) Nef proteins. Overall, 6 (10%) of 60 persons had truncated Nef proteins; of these, 5 were among the 36 asymptomatic subjects (13.9%) and only 1 was among the 24 symptomatic subjects (4.2%) (P =.23). The results of this study document the presence of defective nef genes in HIV-2 infections with a prevalence higher than that previously seen in HIV-1-infected cohorts of long-term nonprogressors or patients with AIDS.