Herpes simplex virus type 1 (HSV-1) is a highly prevalent human herpesvirus which infects approximately two-thirds of individuals worldwide, typically causing vesicular lesions (‘cold sores’) at the orolabial mucosa. The success and lifelong persistence of HSV-1 is partly attributed to the range of immunoevasive mechanisms employed by the virus, which include viral downregulation of classical and non-classical antigen presentation molecules, such as MHC-I, CD1d and MR1. Although HSV-1 infects and replicates at mucocutaneous regions, interactions between the virus and mucosal-associated invariant T (MAIT) cells are largely uncharacterised. Here we report for the first time that HSV-1 is capable of directly infecting peripheral blood MAIT cells and manipulating their effector phenotype. Using spectral cytometry and tetramer-based identification of MAIT cells, we show using an in vitro model of infection that HSV-1 infects distinct primary human MAIT cell subsets (CD4+, CD8+, CD4-CD8-, and CD4+CD8+) and alters their activation profile in response to stimulation via T-cell receptor (TCR)-dependent and TCR-independent pathways. The impact of HSV-1 infection on MAIT cell function is profound and involves the suppression of cell-surface activation markers (CD25, CD69, PD-1), impaired expression of key cytokine receptors IL-18R and IL-12R, and potent inhibition of Th1-associated cytokine production (IFN-γ, TNF). Further, HSV-1 infected MAIT cells display reduced cytotoxic potential, with limited degranulation and perforin/granzyme B expression following stimulation with the MR1 ligand 5-OP-RU and/or cytokines. Across a range of effector outputs, we also find that HSV-1 impacts the functional capacity of ‘bystander’ MAIT cells, which are exposed to HSV-1 infection but remain viral antigen-negative. Collectively, this work reveals MAIT cells are susceptible to functional disruption via infection or exposure to a prevalent human herpesvirus. This adds to our growing understanding of the effects of viral infection on MAIT cell phenotype and function.