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Antigen-induced deoxyribonucleic acid synthesis in mouse lymphocytes. I. The nature and specificity of lymphocyte activation by hemocyanin and dinitrophenyl carrier conjugates.


 The deoxyribonucleic acid (DNA) synthetic response of mouse spleen cell cultures has been studied after stimulation with the soluble antigens, keyhole limpet hemocyanin (KLH) and dinitrophenyl (DNP)-KLH. KLH was capable of inducing mitosis in normal as well as KLH-primed mice, the greater reactivity in primed mice apparently reflecting a specific anamnestic response. The kinetics of KLH antigen-induced DNA synthesis showed an earlier peak response (48 hr) than that induced by the mitogen concanavalin A (72 hr). In contrast, spleen cell cultures from normal mice failed to respond to DNP-KLH while DNP-KLH-primed spleen cells produced a highly significant DNA synthetic response. This response was dose-dependent and required the presence of the intact priming conjugate, since DNP-ovalbumin, DNP-bovine γ-globulin, DNP-lysine, and DNP-ε-amino caproic acid failed to stimulate such DNP-KLH-primed cells. To a great extent, the response was hapten-specific, since dissociated KLH, the form conjugated to DNP, stimulated only low levels of DNA synthesis. Moreover, the DNP-KLH-induced response could be inhibited more than 50% by the presence of the univalent DNP-ligand, DNP-ε-amino caproic acid. Finally, DNP-copolymer of d-glutamic acid and d-lysine, a molecule for which no thymus-derived lymphocytes exist, induced a DNA synthetic response in both normal and DNP-KLH-primed spleen cell cultures, and priming with DNP-KLH failed to increase the level of response to this molecule over that seen in normal spleen cells.