Real-time monitoring of mainstream and social media can inform public health practitioners and policy makers about vaccine sentiment and hesitancy. We describe a publicly available platform for monitoring vaccination-related content, called the Vaccine Sentimeter. With automated data collection from 100,000 mainstream media sources and Twitter, natural-language processing for automated filtering, and manual curation to ensure accuracy, the Vaccine Sentimeter offers a global real-time view of vaccination conversations online. To assess the system’s utility, we followed two events: polio vaccination in Pakistan after a news story about a Central Intelligence Agency vaccination ruse and subsequent attacks on health care workers, and a controversial episode in a television program about adverse events following human papillomavirus vaccination. For both events, increased online activity was detected and characterized. For the first event, Twitter response to the attacks on health care workers decreased drastically after the first attack, in contrast to mainstream media coverage. For the second event, the mainstream and social media response was largely positive about the HPV vaccine, but antivaccine conversations persisted longer than the provaccine reaction. Using the Vaccine Sentimeter could enable public health professionals to detect increased online activity or sudden shifts in sentiment that could affect vaccination uptake.