Urothelial carcinoma of the bladder is a solid tumor entity for which the immunomodulatory agent Bacillus-Calmette Guerin (BCG) has been shown to have substantial efficacy with approximately 90% cure rates in select patients with superficial disease. Immune-based therapies for patients with more invasive disease do not currently exist. We previously showed that invasive urothelial carcinomas express the NY-ESO-1 tumor antigen. Here we evaluated the safety and immunogenicity of a recombinant NY-ESO-1 protein vaccine, which was administered with granulocyte macrophage colony-stimulating factor and BCG as immunologic adjuvants in a cohort of urothelial carcinoma patients. Sixty-two urothelial carcinoma patients were screened for enrollment onto the vaccine clinical trial and 6 patients met all eligibility criteria to receive vaccination. Patients with localized disease underwent surgical removal of their bladders as treatment for their disease. Tumor tissues were tested for NY-ESO-1 expression and eligible patients, shown to have NY-ESO-1 tumors, were vaccinated in the postoperative setting. Peripheral blood samples were analyzed for vaccine-induced antibody and T-cell responses. The vaccine regimen was well tolerated with only mild injection site reactions. NY-ESO-1-specific antibody responses were induced in 5/6 patients whereas CD8 T-cell responses occurred in 1/6 patients and CD4 T-cell responses were found in 6/6 patients. This study demonstrates safety and feasibility of the NY-ESO-1 recombinant protein in combination with BCG and granulocyte macrophage colony-stimulating factor to induce predominantly antibody and CD4 T-cell responses in urothelial carcinoma patients. Induction of higher frequency of CD8 T-cell responses may be possible in clinical trials implementing NY-ESO-1 vaccination in combination with other immunomodulatory agents.