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In humans and in BALB/c mice, immune responses to the hormone insulin use evolutionarily related VHV (human) and VHIX (murine) gene families. To determine if these structural relationships include regulatory elements, BALB/c mice were pretreated with autologous immunoglobulin G (IgG) monoclonal antibodies (mAb) that recognize shared idiotopes on human anti-insulin antibodies and the subsequent immune response to human insulin assessed. One mAb, Id227, was found to augment and accelerate the insulin response by inducing a human idiotype that is expressed on both insulin-binding and non-insulin-binding BALB/c antibodies. Analysis of VH gene utilization by Id227 shows that it expresses a VHIX gene similar to that of anti-insulin mAb 125, but the anti-Id has no anti-insulin activity. Using DNA amplification, four germ-line VHIX genes were isolated from BALB/c liver DNA and sequence analysis shows that the anti-insulin and anti-Id are derived from the same germ-line gene. Consistent with its role as a regulatory idiotype, IgG Id227 entirely preserves germ-line sequence in the complementary determining regions and contains only three mutations in framework regions. These studies show that both structural and regulatory features of immune responses to conserved self antigens extend beyond species boundaries.