Studies of Human Papillomavirus (HPV) knowledge among university students have previously been conducted, typically to the exclusion of faculty and staff. The study aim was to assess HPV knowledge among university students, staff, and faculty in the state of Indiana during 2016, 2019, and 2022. The three samples comprised a total of 1275, 1056, and 403 students, staff, and faculty respectively during 2016, 2019, and 2022. Ages ranged from 18 to 70 + each year of the study. A 27-item survey was administered including sociodemographic questions and quantitative HPV knowledge questions were modified from a previously validated instrument. Descriptive statistics characterized the study participants and differences in scores were reported. Among the overall sample who answered receiving the vaccine question (n = 1917), 39% had received the HPV vaccine. HPV knowledge significantly increased among students from 2016 to 2019 but decreased in 2022. Among staff, there was a significant knowledge difference between 2016 and 2022, but no significant knowledge difference among the faculty. About 86% of the total sample did not know that HPV usually does not need any treatment. Further, participants correct knowledge scores were less than average on the following items: HPV can cause throat cancer (49%); HPV can be transmitted by skin-to-skin contact (42%). More education and campus-wide efforts need to be implemented to increase awareness and accurate HPV-knowledge. Targeted and individual messaging for students, staff, and faculty may be a way to increase awareness of HPV and improve HPV knowledge.