Background: HIV eradication may require synergistic interventions that purge latent reservoirs and increase the HIV-specific cellular immune response. DNA prime followed by recombinant adenovirus 5 boost vaccination (HIV-rAd5) has been shown to induce strong CD8+ cytotoxic T lymphocytes responses in HIV-infected and uninfected individuals. Methodology: Randomized, controlled, non-comparative clinical trial of ART intensification (raltegravir and maraviroc (RAL/MVC)) with or without HIVrAd5. We included patients having: ≥3 years of HIV-1 RNA ≤500 copies/mL, HIV-1 RNA <40 copies/mL ≥one year, current CD4+ cell count ≥350/mm3, HIV-1 DNA between 10 and 1,000 copies/106 PBMCs and serum Ad5 neutralizing antibody titer ≤ 250. After a lead-in of RAL/MVC intensification for 8 weeks, patients were randomized (1:1) to continue intensification with HIV-rAd5 vaccination [DNA prime at W8-W12-W16 and rAd5 boost at W32 (RAL/MVC/HIV-rAd5 arm)] or continue intensification alone (RAL/MVC arm) for 48 weeks. The primary end-point was >0.5 log10 decrease in HIV-1 DNA at 56 weeks. Secondary end-points included changes in rectal tissue HIV-1 DNA, immunologic changes in peripheral blood, and safety. Results: We enrolled 28 patients (14 in each arm) on suppressive ART with the following median baseline characteristics: 636 CD4+ cells/mm3, 672 CD8+ cells/mm3, 170 HIV-1 DNA copies/106 PBMCs, 13 years on ART, and 2.6 years with HIV-1 RNA<40 copies/mL. At week 56, one patient in the RAL/ MVC alone arm reached the primary endpoint with a 0.55 log10 decrease in HIV-1 DNA from 156 (W0) to 44 (W56) DNA copies/106 PBMCs. Mean PBMC and rectal tissue cell-associated HIV-1 DNA levels did not significantly change after RAL/MVC intensification with or without HIV-rAd5 injections (see table). Peripheral blood CD4+ and CD8+ cell counts did not significantly change from baseline to W56 in either arm (see table). RAL/MVC intensification and HIV-rAd5 vaccination were well tolerated and there were no serious SAEs related to study treatment. Conclusions: RAL/MVC intensification with HIV-rAd5 vaccination did not reduce the total HIV DNA reservoir in either peripheral blood or rectal tissue. RAL/MVC intensification alone decreased the HIV-1 reservoir by over 0.5 log DNA copies/106 PBMCs in one patient.