Abstract A significant challenge to HIV eradication is the elimination of viral reservoirs in germinal center (GC) resident T follicular helper cells. Therapeutic interventions that target viral reservoirs should induce anti-viral CD8 T cells that home to GC. We and others recently described a unique population of CD8 T cells that express the chemokine receptor CXCR5 (required for homing to GC) during chronic HIV/SIV infections. A critical question that remains to be addressed is if follicular homing CD8 T cells can be induced by vaccination and if so how will they impact control of HIV. Here we show that vaccination of rhesus macaques (RM) with DNA/MVA (modified vaccinia Ankara) SHIV vaccine induces low levels of vaccine-specific CXCR5+ CD8 T cells in blood. However, adjuvanting the DNA vaccine with CD40L dramatically enhanced the induction of CXCR5+ SHIV-specific CD8 T cells up to 10-fold with a frequency of about 5% of total CD8. These CXCR5+ CD8 T cells expressed higher levels of cytolytic molecules perforin and granzyme-B with higher proliferative capacity (ki-67) compared to CXCR5-subset. Following a pathogenic SHIV infection, the CXCR5+ CD8 subset expanded rapidly. Importantly, the frequency of SHIV-specific CXCR5+ CD8 T cells post vaccination and post infection showed a significant inverse correlation with viral load (p=0.003, R=−0.66), these associations were not observed with CXCR5-CD8 T cells. These data demonstrate that CXCR5+ CD8 T cells can be induced by vaccination, their induction can be augmented by CD40L, and the vaccine-induced CXCR5+ CD8 T cells contribute to control of pathogenic immunodeficiency virus infection. These findings have important implications for developing HIV cure strategies.