In this paper, we present the interference neutralization technique for two-hop multiple-input multiple-output (MIMO) relay systems. It enables multiple MIMO transmitters (sources) to simultaneously transmit independent data streams to their MIMO receivers (destinations) without mutual interferences, thereby improving spectral efficiency of the systems. To neutralize the mutual interferences using multiple amplify-and-forward (AF) MIMO relays, we establish the sufficient condition for the antenna configuration in the MIMO relay networks, and provide a filter design technique for the AF MIMO relays. The proposed method increases sum rates of the systems linearly with the number of transmitters participating in simultaneous transmission. To improve the sum rates further, this method is combined with transmit power allocation using the water-filling algorithm. In addition, it is shown that by employing the minimum number of relays required to meet the sufficient condition, the system cost for the proposed method can be reduced without compromising the sum rate performance severely. Finally, simulation results successfully demonstrate that by exploiting radio resources such as frequency and time efficiently, the proposed method achieves a higher sum rate than the existing techniques based on interference avoidance. (C) 2016 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.