Background: Despite the introduction of the human papillomavirus (HPV) vaccination as a preventive measure in 2006 for cervical and other cancers, uptake rates remain suboptimal, resulting in preventable cancer mortality. Social media, widely used for information seeking, can influence users’ knowledge and attitudes regarding HPV vaccination. Little is known regarding attitudes related to HPV vaccination on Reddit (a popular news aggregation site and online community), particularly related to cancer risk and sexual activity. Examining HPV vaccine–related messages on Reddit may provide insight into how HPV discussions are characterized on forums online and influence decision making related to vaccination. Objective: We observed how the HPV vaccine is characterized on Reddit over time and by user gender. Specifically, this study aimed to determine (1) if Reddit messages are more related to cancer risks or sexual behavior and (2) what other HPV vaccine–related discussion topics appear on Reddit. Methods: We gathered all public Reddit comments from January 2007 to September 2015. We manually annotated 400 messages to generate keywords and identify salient themes. We then measured the similarity between each comment and lists of keywords associated with sexual behavior and cancer risk using Latent Semantic Analysis (LSA). Next, we used Latent Dirichlet Allocation (LDA) to characterize remaining topics within the Reddit data. Results: We analyzed 22,729 messages containing the strings hpv or human papillomavirus and vaccin. LSA findings show that HPV vaccine discussions are significantly more related to cancer compared with sexual behavior from 2008 to 2015 (P .05). LDA analyses demonstrated that although topics related to cancer risk and sexual activity were both frequently discussed (16.1% and 14.5% of word tokens, respectively), the majority of online discussions featured other topics. The most frequently discussed topic was politics associated with the vaccine (17.2%). Other topics included HPV disease and/or immunity (13.5%), the HPV vaccine schedule (11.5%), HPV vaccine side effects (9.7%), hyperlinks to outside sources (9.1%), and the risks and benefit of HPV vaccination (8.5%). Conclusions: Reddit discourse on HPV vaccine encompasses a broad range of topics among men and women, with HPV political debates and cancer risk making up the plurality of the discussion. Our findings demonstrated that women and men both discussed HPV, highlighting that Reddit users do not perceive HPV as an issue that only pertains to women. Given the increasing use of social media as a source of health information, these results can inform the development of targeted online health communication strategies to promote HPV vaccination to young adult users of Reddit. Analyzing online discussions on Reddit can inform health communication efforts by identifying relevant, important HPV-related topics among online communities. [JMIR Public Health Surveill 2019;5(1):e12480]