Antigen-Induced Deoxyribonucleic Acid Synthesis in Mouse Lymphocytes

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Abstract
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.

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