An important aspect of successful emotion regulation is the ability to adjust emotional responses to changing environmental cues. Difficulties with such adaptation may underlie both marked symptoms of behavioral inflexibility and frequent severe anxiety in the autism spectrum disorders (ASDs). Thirty children and adolescents diagnosed with ASD and 29 age- and intelligence quotient-matched controls completed a reversal learning paradigm following partial reinforcement Pavlovian fear conditioning, using a surprising air puff as the unconditioned stimulus. After initial reversal of cue contingencies, where a previously safe cue now predicted the air puff threat, the control group but not the ASD group responded more strongly to the new threat cue. The ASD group showed evidence for reversal learning only during later trials. Reversal learning in the ASD group was significantly negatively correlated with everyday symptoms of behavioral inflexibility but not with everyday anxiety. Understanding shared associations between inflexibility, anxiety, and autism, with regard both to clinical symptoms and neurobiological mechanisms, can provide important markers for better characterizing the substantial heterogeneity across the autism spectrum. Autism Res 2012, 5: 398–406. © 2012 International Society for Autism Research, Wiley Periodicals, Inc.