Mucosal-associated invariant T (MAIT) cells are a subset of pro-inflammatory innate-like T cells that are highly abundant in human tissues and recognize small molecule metabolite antigens presented by the MHC class-I-like molecule, MR1. MAIT cells are found in almost all mammals, whereby the MAIT-MR1 axis is highly conserved, suggesting an important physiological function linked to metabolite antigen sensing. A well-characterised function of MAIT cells is protection during bacterial and fungal infections upon recognition of the MR1-presented antigen 5-OP-RU, a riboflavin-synthesis-derived metabolite. Whether MAIT cells contribute to immunity from riboflavin-deficient parasites upon recognition of a novel antigen is unclear. Because of the large frequency of MAIT cells in the human liver and the well-established protective capacity of CD8+ T cells during liver-stage malaria, we sought to characterise the role of MAIT cells during liver-stage malaria using a mouse model of liver-stage infection. Current results indicate that two days following infection with Plasmodium berghei, the liver parasite burden of MR1-/- C57BL/6 mice is significantly higher than that of MAIT cell-proficient wild-type mice. This suggests that MAIT cells may contribute to protection. In vitro, when PMA-differentiated THP-1 cells were co-incubated with Plasmodium falciparum or P. berghei sporozoites and MAIT cells enriched from healthy human blood, MAIT cells displayed a polyfunctional pro-inflammatory cytokine and cytotoxic response. Assays with MR1 blockade or MR1 overexpressing and knock-out THP-1 cells, demonstrated activation was dependent on antigen presentation by MR1. As Plasmodium spp. lack the riboflavin biosynthesis pathway, this data alludes to the existence of a novel Plasmodium liver-stage-infection-derived MAIT cell antigen. Consequently, our findings suggest MAIT cells play a protective role in the immune response to liver-stage malaria and have the potential to provide the conceptual basis for a broad-spectrum MAIT cell-based malaria vaccine for Plasmodium infection.