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Mass gatherings such as the Olympic Games pose unique challenges for interorganizational collaboration. Such events often bring together organizations that collaborate irregularly or have never engaged in joint working. They involve interaction and collaboration among multiple and diverse agencies aiming at delivering a service to a large clientele, which can often prove challenging. This study used the 2012 London Olympic Games as the empirical setting to examine the interagency collaboration among the multiple and diverse public health and safety organizations involved in one of the world's largest mass gatherings. A single, holistic and exploratory case study design was used and data were collected before, during and after the Games through 39 semi-structured interviews with key informants, direct observations of field exercises and documentary analysis. Data collection commenced in May 2011, 14 months before the actual Games, and was completed in October 2012, two months after the completion of the Games. Template analysis was used to thematically analyze the interviews' transcripts, the fieldnotes from observations and the documents. Findings discuss interagency collaboration in mass gatherings along three main activity domains: leadership, communication and learning. In each domain, a number of challenges and facilitators emerged influencing interagency collaboration. Regarding the leadership domain, the lack of engagement of the leading organization and the ambiguous interoganizational decision-making processes negatively influenced collaboration. Shared micro-level leadership and the use of interorganizational linkages enabled collaborative working. Experienced positional leaders of each organization enabled the decision-making process at the interagency operational level by exercising a range of interpersonal leadership capabilities including flexibility and the ability to negotiate. Codified frameworks at the organizational level also provided leaders with common ground to assist them manage the complex interorganizational processes. Within the second domain, the complex intraorganizational structure of the involved agencies and the high density of information transmitted were associated with a dysfunctional communication experience. Findings revealed that the crafting of boundary-spanning roles and intense face-to-face interaction positively contributed to interagency collaboration. Online information systems and formal intersectoral dissemination of reports were essential in gaining common situational awareness. The implicit cultural rules in the form of communication etiquette shaped how interorganizational collaboration was perceived. Finally, sharing the acquired knowledge was a necessary step to create an enabling collaborative environment among interacting organizations. Experiential learning was identified as a significant factor which helped promote joint understanding and partnership work. Informal interpersonal exchanges and formal knowledge transfer activities facilitated knowledge sharing across interorganizational boundaries, helping to break down silos. The study outlines challenges and strategies that shaped interagency collaboration in the context of mass gatherings. Practical implications arising from this study inform the ways organizers of mass gatherings, public health and safety agencies and professionals can engage in effective partnerships and joint working.