Bone marrow (BM)-FMS-like tyrosine kinase 3 (Flt3) ligand-murine dendritic cells (DC) in response to group A streptococcal (GAS) lipopeptide vaccines containing an analogue of a Toll-like receptor (TLR) 2 agonist showed significant up-regulation in expression of DC maturation markers CD40 and CD80 when compared to unstimulated controls. There were significant increases in MHC class II, CD40, CD80 and CD86 expression on classical DC and plasmacytoid DC after in vivo administration or in vitro stimulation of BM-granulocyte macrophage colony stimulating factor (GMCSF)-DC with lipopolysaccharide but not lipopeptide GAS vaccines. Our results indicate that an LCP-GAS vaccine induced phenotypic maturation of BM-Flt3-DC but not BM-GMCSF-DC.