During the Covid-19 pandemic, many governments promoted the adoption and development of telework to reduce some of the consequences of the current health crisis on the economy and favor social distancing. The aim of this web-based cross-sectional study was to assess the consequences of the Covid-19 pandemic on job organization, exploring the effects of lockdown measures on the psychological distress and perceived well-being of workers experiencing telework. A web-based cross-sectional survey has been used to collect data. The participants answered the questionnaire from April 1 to April 30, 2020. The questionnaire consisted of three sections, which investigated: 1) demographic and occupational variables, 2) lifestyle and habits variables, 3) psychological distress and perceived well-being. The General Health Questionnaire (GHQ-12) has been used to evaluate psychological distress and the 5-item World Health Organization Well-Being Index (WHO-5) to explore subjective well-being. Psychological distress was associated with educational level, with habits, and with reporting poor well-being. Poor well-being was associated with a higher job demand during pandemic, lifestyle and habits variables, and psychological distress. This is one of the first studies exploring the consequences of the COVID-19 pandemic and lockdown measures on the perceived well-being and psychological distress of workers experiencing telework. It is mandatory to pay more and more attention to the mental health of teleworkers, considering the increasing diffusion and adoption of this type of work organization.