Injuries and fatalities for vulnerable road users, especially bicyclists and pedestrians, are on the rise. To better inform design for vulnerable road users, we need to evaluate how bicyclist and pedestrian behavior and physiological states change in different roadway design and contextual settings. Previous research highlights the advantages of using immersive virtual environments (IVEs) in conducting bicyclist and pedestrian studies. These environments do not put participants at risk of injury, are low cost compared to on-road or naturalistic studies, and allow researchers to fully control variables of interest. In this paper, we propose a framework, Omni-Reality and Cognition Lab Simulator (ORCLSim), to support human sensing techniques within IVEs to evaluate bicyclist and pedestrian physiological and behavioral changes in different contextual settings. To showcase this framework, we present two case studies, where pilot data from five participants’ physiological and behavioral responses in an IVE setting are collected and analyzed, representing real-world roadway segments and traffic conditions. Results from these case studies indicate that physiological data are sensitive to road environment changes and real-time events in the IVE, especially changes in heart rate and gaze behavior. In addition, our preliminary data indicate participants may respond differently to various roadway settings (e.g., signalized vs. unsignalized intersections). By analyzing these changes, future studies can identify how participants’ stress level and cognitive load are impacted by the surrounding environment. The ORCLSim system architecture is a prototype that can be customized for future studies in understanding users’ behavioral and physiological responses in virtual reality settings.