Chronic viral infections such as HIV are responsible for a significant global burden of disease and have proven to be difficult to target with effective vaccines. While the immune defects in chronic infectious diseases are multifactorial, a key problem is the dysregulation of the humoral immune response in these patients. Memory B cell subsets provide our immune system with the ability to adapt to different antigens and provide protection upon secondary exposure to pathogens. During chronic viral infection, there is a skewing of the immune response towards short-lived plasmablasts and an expansion of atypical memory B cells. These cells have a reduced ability to produce neutralising antibody responses, along with increased expression of inhibitory receptors (FcRL4 and PD-1) and poor proliferative capacity and may play a role in allowing the pathogens to evade clearance by the immune system. There is no established in vivo animal model for atypical memory B cell formation in the context of chronic viral infection and little is known about the requirements for the formation of memory B cell subsets in viral infection. We used acute and chronic strains of lymphocytic choriomeningitis virus (LCMV) in mice in conjunction with a fluorochrome-conjugated B cell tetramer specific for the LCMV nucleoprotein in order to examine memory B cell subset formation following infection and determine whether expansion of atypical memory B cells is associated with impaired immune memory. We found increased germinal centre B cells in chronic infection, compared to acute infection, over time. However, fewer of these were antigen-specific compared to those produced in acute infection. Antigen-specific B cells in vivo had increased expression of CXCR3, PD-1, and reduced expression of CD21, comparable to the phenotype of human atypical memory B cells. CD138+ B cells were increased in chronic infection compared to acute, indicating a dysregulation of differentiation into plasma cells. Identifying the requirements and functions of different memory B cell subsets in acute and chronic infectious states will provide insight into the mechanisms driving B cell subset formation, and may allow for effective future vaccine development.