Human papillomavirus (HPV) is a highly widespread virus which is responsible for one of the most common sexually transmitted infections. Two main preventative strategies exist: anti-HPV vaccination and cervical screening. Health-care workers play a key role in promoting public health campaigns; however, vaccine hesitancy is an often under-recognized challenge. To investigate the overall knowledge of HPV and HPV-related issues, as well as the attitudes and practices of health professionals towards recommending the anti-HPV vaccine, an ad hoc knowledge, attitudes, and practices (KAP) questionnaire was developed and validated in a sample of 139 Israeli pediatricians, gynecologists, and internal medicine doctors. The KAP questionnaire was found to be psychometrically valid and sound (with an rKR-20 coefficient of 0.74 for the second part and a Cronbach’s alpha of 0.85 for the third part). Furthermore, the present study confirmed the importance of health-care providers in recommending the immunization practice. Parents that had been strongly advised by health-care providers to vaccinate their children accepted immunization for their girls (odds ratio (OR) 1.09 (95% CI 1.04–1.14)) and boys (OR 1.06 (95% CI 1.02–1.10)), had a lower probability of deciding to postpone the immunization appointment (OR 0.81 (95% CI 0.66–0.98)), had fewer doubts and concerns about the vaccine (OR 0.69 (95% CI 0.54–0.89)), and had a lower probability of refusing the vaccination (OR 0.93 (95% CI 0.86–0.99)). Interestingly, the use of new, emerging tools such as ad hoc websites, applications, and other interactive devices reduced vaccine hesitancy (OR 0.90 (95% CI 0.82–0.99)) and concerns about the side-effects of the vaccine (OR 0.92 (95% CI 0.86–0.99)). However, among Israeli health-care workers, knowledge was generally moderate, with updated information lacking in about 30% of surveyed health-care providers and approximately 20% of them not recommending the anti-HPV vaccine among boys. This study has practical implications for policy- and decision-makers in that they should be aware of the overall level of knowledge among health-care workers and should implement ad hoc educational interventions to address gaps in knowledge and help medical providers routinely recommend the anti-HPV vaccine both to male and female children and adolescents.