About 20% of health care workers (HCWs) acquired influenza every year, continuing to work and encouraging virus spreading. Vaccination coverage during last 11 seasons among HCWs of the University Hospital of Palermo were analyzed, assessing the impact of tailored communicative and informative strategies and evaluating knowledge and attitudes for vaccination acceptance or refusal. Among initiatives organized to increase HCWs awareness and vaccine acceptance were: dedicated web and social-media informative pages, mandatory informed dissent, “on site” vaccination. Moreover, reasons for influenza vaccination acceptance or refusal were evaluated. Vaccination coverage raise from a mean value of 5% (from 2010/2011 to 2014/2015) to 27% in 2017/2018 influenza season (chi-square for trend: 857, p < 0.001). Mean age of vaccinated HCWs went down from 48.1 (±15.7) in 2013/2014 to 41.6 (±14.5) in 2017/2018. Risk perception of contracting or transmitting influenza virus were main reasons for acceptance. Otherwise, fear of adverse reaction and a substantial lack of perception of spreading influenza among patients were reported as main refusal motives. Communicative and informative strategies undertaken at the University Hospital of Palermo produced a significant increase of vaccination adherence, reaching rates consistent with mean European coverages, although still far from 75% recommended by Public Health Authorities.