The purpose of this study was to assess the knowledge and attitudes about human papilloma virus (HPV) and cervical cancer, and the acceptability of HPV vaccine among students, parents and teachers in secondary schools in Bangkok, Thailand. We conducted a school-based cross-sectional study at four public secondary schools in Bangkok. A total of 644 students aged 12-15 years, 664 parents and 304 teachers were recruited into the study. Data were collected by self-administered questionnaires. The percentages of students, parents and teachers who were willing to be vaccinated were 26, 49 and 43%, respectively. Forty-one percent of parents wanted their children to be vaccinated. Students, parents and teachers had a moderate knowledge of HPV, cervical cancer and the HPV vaccine with mean scores of 6.91 (SD = 1.75), 6.82 (SD = 1.88), and 6.70 (SD = 1.89), respectively. The attitudes of students, parents, and teachers were fair with scores of 3.46 (SD = 0.41), 3.52 (SD = 0.43), and 3.46 (SD = 0.47) out of 5, respectively. Twenty-nine percent of students and 36% of parents were willing to pay USD 14.3-28.5 per dose for the quadrivalent vaccine; 33% of teachers were willing to pay < USD 14.3 per dose for the quadrivalent vaccine. This study is the first study to report the knowledge, and attitudes and acceptability of HPV vaccination in Thailand. The findings suggest the willingness to pay was relatively low and related to the price, while knowledge and attitudes regarding the importance of the HPV vaccine were fair particularly among parents and teachers. Greater effort may be needed to educate people regarding the cost and benefits of HPV vaccination before it would be more acceptable to parents, teachers and students in Thailand.