Background: Nurses may be unwilling to accept HIV-infected colleagues who can continue towork with patients if the provider-to-patient transmission risks have been appropriately assessed. Aims: To assess the factors associated with nurses' willingness to accept HIV-infected colleagues as coworkers. Design: Descriptive and cross-sectional. Methods: An anonymous online survey targeting Japanese nurses working in hospitals or clinics (n=992). Logistic regression analysis was used to evaluate factors associated with their willingness to accept HIV-positive colleagues before or after a risk assessment. Results: Respondents who avoided contact with, and expressed discriminatory views about, HIV-infected colleagues were less willing to accept them. After undertaking an appropriate risk assessment, a high level of knowledge regarding HIV transmission increased the likelihood of acceptance towards HIV-infected colleagues. Conclusions: The use of risk assessment may increase nurses' willingness to accept HIV-infected colleagues, particularly among co-workers with a high level of knowledge about HIV.