In this study, we conducted a GWAS of HIV-1 acquisition among a high-risk sample of 3,136 IDUs from the Urban Health Study (UHS). In addition to both cases and controls being IDUs, HIV- controls were frequency-matched to HIV+ cases on a number of exposure risks (e.g., sexual risks)—enhancing detection of genetic contributions to differences in HIV status. We tested for independent replication in the Women’s Interagency HIV Study (WIHS, N = 2,533) and examined gene expression data to link the replicated novel SNP association with HIV acquisition to known mechanisms affecting viral replication and infectivity during acute HIV exposure.