The exact specifiicity of anti-DNP antibody produced by Hartley guinea pigs immunized with a series of defined α,DNP and ϵ,DNP-oligolysines was studied by fluorescence quenching. All responder animals made anti-DNP antibody which recognized the precise chain length, ± 1 lysyl residue, of the DNP-oligolysines used to induce the immune response as measured by an increase in binding energy (–ΔF°) for that antigen. The ability of the immune system to detect the smallest possible change in oligolysine chain length suggests that the anti-hapten antibody-forming cell possesses a highly specific recognition system for carrier conformation. When DNP-oligolysines are incorporated in an adjuvant containing M. tuberculosis H37Rv, both responder and nonresponder produce anti-DNP antibody, but only the responder develops delayed skin sensitivity. In addition to their failure to develop delayed hypersensitivity, nonresponders produced anti-DNP oligolysine antibody which did not show the increase in –ΔF° for the immunizing antigen characteristic of responder antibody. These observations support a local environment hypothesis for antigen recognition at the level of the anti-hapten antibody-forming cell and suggest that the polylysine gene exerts its control at the same cell.

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