Vaccines are among the most important public health achievements of the last century; however, vaccine awareness and uptake still face significant challenges and the COVID-19 pandemic has only exacerbated this phenomenon. Vaccine Literacy (VL) is the ability to find, understand and judge immunisation-related information to make appropriate immunisation decisions. A cross-sectional study on a sample of 3500 participants, representative of the Italian adult population aged 18+ years, was conducted in Italy in 2021. A validated questionnaire, including sections on health literacy (HL), sociodemographic characteristics, risk factors, and lifestyles of respondents, was used. VL was measured by four items (item 19, 22, 26 and 29) of the HL section. While 67.6% of the respondents had a “good” (47.5%) or “sufficient” (20.1%) level of VL, 32.4% had “limited” VL levels. Although the overall VL level was quite high, many participants reported difficulties in dealing with vaccination information, particularly those with a lower educational level, those living in southern and insular regions of Italy, those with greater financial deprivation and those with a migration background. Improving VL in Italy should be a top priority in the political agenda, with special regard to socially and geographically disadvantaged communities.