One of the most striking features of exposure to low doses of mercury in mice is the high-titer haplotype-linked anti-nucleolar (ANoA) autoantibody response. Mice of H-2(s) haplotype have been high responders, while H-2(b) mice have been low. This pattern has been attributed to the class II molecule itself, but the poor response of F1 crosses between high and low responders raised the possibility that the anti-fibrillarin specificity was actually due to a closely linked dominant negative gene. To test the role of class II explicitly, F1 crosses between congenic B6.SJL (H-2(s)) and C57BL/6 (H-2(b)) mice with a targeted deletion of I-AbAbeta were generated, creating mice heterozygous for all MHC loci, but expressing only I-As. In comparison with B6.SJL, no diminution of ANoA titers was found, proving that I-As itself was responsible for susceptibility and I-Ab for downregulation. Unlike I-A, expression of the I-E class II molecule could not downregulate the response in an otherwise susceptible mouse. These results suggest a complicated role for class II in the regulation of a novel, environmentally induced autoimmune response.