question: Background: The immunization of healthcare workers (HCWs) plays a recognized key role in prevention in the COVID-19 pandemic: in Italy, the vaccination campaign began at the end of December 2020. A better knowledge of the on-field immune response in HCWs, of adverse effects and of the main factors involved is fundamental. Methods: We performed a study on workers at a nursing home in Northern Italy, vaccinated in January–February 2021 with two doses of the BNT162b2 vaccine four weeks apart, instead of the three weeks provided for in the original manufacturer protocol. One month after the second dose, the serological titer of IgG-neutralizing anti-RBD antibodies of the subunit S1 of the spike protein of SARS-CoV-2 was determined. The socio-demographic and clinical characteristics of the subjects and adverse effects of vaccination were collected by questionnaire. Results: In all of the workers, high antibody titer, ranging between 20 and 760 times the minimum protective level were observed. Titers were significantly higher in subjects with a previous COVID-19 diagnosis. Adverse effects after the vaccine were more frequent after the second dose, but no severe adverse effects were observed. Conclusions: The two doses of the BNT162b2 vaccine, even if administered four weeks apart, induced high titers of anti-SARS-CoV-2 neutralizing IgG in all the operators included in the study.