Antigen Recognition and the Immune Response: the Capacity of L-Tyrosine-Azobenzenearsonate to Serve as a Carrier for a Macromolecular Hapten

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Abstract
The low molecular weight compound L-tyrosine-azobenzenearsonate (RAT) induced immunity in guinea pigs, as evidenced by delayed cutaneous hypersensitivity and stimulation of DNA synthesis by lymphoid cells in culture. Two different conjugates of RAT and a poly-γ-D-glutamic acid hapten of m.w. 35,000 were prepared. In one, an average of nine molecules of RAT were linked directly to the PGA backbone [PGA-(RAT)9]. In the other, RAT was joined to PGA by means of 6-aminocaproyl (SAC) hydrocarbon chains that extended the RAT groups out from the PGA backbone by a maximum of 10 Å [PGA-(SAC-RAT)12]. Immunization with the two conjugates gave rise to anti-PGA antibody, PGA-(SAC-RAT)12 being somewhat more effective in that regard, and all animals developed delayed hypersensitivity to RAT but not to PGA. Lymphoid cells from animals sensitized to PGA-RAT conjugates demonstrated specificity for the homologous conjugates, assessed by the rate of incorporation of 14C-thymidine into DNA. Prior immunization with RAT potentiated the response to PGA following administration of the conjugates. Thus, it appears that a molecule with an m.w. of about 400 was recognized by antigenreactive cells and could serve as a carrier for a hapten with an m.w. of 35,000.

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