Magneto-rheological (MR) fluid dampers can be used to reduce the traffic induced vibration in highway bridges and protect critical structural components from fatigue. Experimental verification is needed to verify the applicability of the MR dampers for this purpose. Real-time hybrid simulation (RTHS), where the MR dampers are physically tested and dynamically linked to a numerical model of the highway bridge and truck traffic, provides an efficient and effective means to experimentally examine the efficacy of MR dampers for fatigue protection of highway bridges. In this paper a complex highway bridge model with 263 178 degrees-of-freedom under truck loading is tested using the proposed convolution integral (CI) method of RTHS for a semiactive structural control strategy employing two large-scale 200 kN MR dampers. The formation of RTHS using the CI method is first presented, followed by details of the various components in the RTHS and a description of the implementation of the CI method for this particular test. The experimental results confirm the practicability of the CI method for conducting RTHS of complex systems.