Young adults with autism spectrum disorder (ASD) are attending university in increasing numbers. The importance of acceptance from peers and integration into the university have been recognized as key factors for success. We examined university students’ knowledge and attitudes towards students with ASD, underlying factors that contributed to such attitudes, and whether attitudes changed across two cohorts 5-years apart. The later cohort demonstrated greater knowledge and more positive attitudes toward students with ASD compared to students in the first cohort. However, knowledge was not found to be a significant predictor of attitudes and many students who were knowledgeable about ASD still reported negative attitudes toward participating in university and classroom based activities with students with ASD.