Influenza viruses circulate annually in humans, accumulating mutations in the viral genome. In real-world conditions, people develop an immune history from multiple influenza virus infections and vaccinations, yet most experimental studies utilize na?ve hosts to investigate viral pathogenesis or vaccine efficacy; a scenario that does not reflect actual human exposure and immune responses. Here, we report that mice previously infected with the mouse-adapted H1N1 virus mounted an increased antibody response after vaccination with the split virion quadrivalent seasonal influenza vaccine. Antibodies targeting the H1 antigen component of the vaccine in H1N1 preimmune mice had titers peaking at a log2 HAI titer of 8 versus 2 in na?ve-vaccinated mice at 14 days post-vaccination, whereas H3N2 pre-immune mice showed no improved vaccine responses. Increased total serum class-switched IgG subclass concentrations in the H1N1 preimmune-vaccinated mice accompanied protection from weight loss and mortality when these mice were challenged with an H1N1 pandemic 2009 strain. Temporal sampling and analysis of RNA from mediastinal lymph nodes from preimmune mice by RNA-seq at time points following vaccination revealed distinct regulation of lymphocyte and germinal center-specific genes, including Bcl6, an essential gene for germinal-center-dependent B-cells. Taken together, H1N1 A/FM/1/1947 preimmunity led to a more efficient vaccine response accompanied by altered expression of germinal center-associated genes in the mediastinal lymph node, with implications for future vaccine design.