A major challenge in the development of vaccines against retroviruses is the induction of neutralizing antibodies since they prevent infection of the cells where the virus may persist. The transmembrane envelope (TM) protein contains highly conserved domains and seems to be a suitable target. To study whether vaccinating with a TM protein of a retrovirus could protect from infection in vivo, cats were immunized with the TM protein p15E of feline leukemia virus (FeLV) and subsequently challenged. For the first time we show that immunization with a retroviral TM protein prevented antigenemia. The level of neutralizing antibodies after the boost immunization correlated with the outcome of FeLV infection.