In order to improve the efficacy of current vaccine candidates against HIV/AIDS, we sought to strengthen the induction of immune responses via simultaneous in vivo mobilization of dendritic cells using a chimeric Flt-3 and G-CSF receptor agonists (ProGP). We investigated ProGP treatment in combination with two DNA immunizations encoding HIV-Env89.6, SIV-Gag proteins to increase the priming of immune responses. Administration of this Flt-3/G-CSF chimera elicited marked increases in numbers of both plasmacytoid and myeloid dendritic cells. However, there was no increase seen in T-cell responses either directly following the DNA immunization or after further boosting with MVA vectors expressing HIV-Env89.6p, SIV-Gag. After challenge with SHIV89.6p all animals became infected and no differences were seen between the ProGP treated versus the control group with regard to plasma virus load or CD4 T-cell count. We conclude that besides mobilization of dendritic cells additional stimuli to induce dendritic cell maturation may be needed for avid boosting of antigen specific immune activation.