BackgroundRecent rises in the incidence of vaccine-preventable illnesses and suboptimal vaccine acceptance are considered a consequence of accumulating misinformation. Evidence-based approaches to patient-provider communication are key to addressing vaccine hesitancy. ObjectivesThe aim of this study was to assess vaccination attitudes and foundational knowledge among healthcare professions students. MethodsA 72-item survey was developed to assess vaccine attitudes and knowledge about vaccination among health professions students. The survey incorporated 14 demographics questions, 41 attitude questions, and 17 knowledge questions. 16 of the attitude questions, derived from a set of core questions used to diagnose vaccine hesitancy from the WHO Strategic Advisory Group of Experts on Immunization (SAGE) Vaccine Hesitancy Matrix, were analyzed together to derive a vaccine acceptance score. Results295 anonymous survey responses were collected between July 2019 and November 2020. Respondents represented students enrolled in medical, dental, pharmacy, optometry, and biomedical science health professions programs. Respondents scored 82.0% ± 0.8% (mean ± standard error of the mean) on questions that gauged vaccine acceptance. The mean vaccine acceptance score was 85.4% ± 1.0% for medical students and 88.0% ± 1.6% for biomedical science students. The mean knowledge score across all programs was 67.7% ± 1.1%. The greatest proficiency in knowledge scores was seen amongst medical students (79.0% ± 1.3%). ConclusionsAmongst the different health professions, students in the fields of medicine and biomedical sciences had the highest levels of vaccine acceptance attitudes and knowledge. The vaccine acceptance score can be utilized by health professions educators to guide vaccine education for future health professionals to better prepare them to address vaccine hesitancy and educate patients on vaccination.