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Ionophorous Activity and Murine B Lymphocyte Mitogens

From the Laboratory of Microbiology and Immunology, National Institute of Dental Research and the Laboratory of Theoretical Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014

Abstract

The relationship between ionophorous and B cell mitogen activity has been investigated. Most known ionophores were nonmitogenic for mouse spleen cells. In addition, when tested in a bilayer lipid membrane (BLM) apparatus, most types of B cell mitogens were nonionophorous.

However, excitability-inducing material (EIM), a high m.w. polymeric protein, which is a channel-forming ionophore, was a potent mitogen for mouse B lymphocytes. Similarly, keyhole limpet hemocyanin (KLH), a high m.w. polymeric protein, which is a B cell mitogen, is a channel-forming ionophore. The mitogenic activities of these two compounds were not due to contamination with endotoxin since they produced weak or absent responses in the limulus lysate clotting and rabbit pyrogenicity assays, and were also mitogenic for spleen cells of endotoxin-low responder C3H/HeJ mice.

Both the mitogenic and ionophorous activities of EIM and KLH were dependent on their polymeric structure since dissociation of these compounds into monomeric subunits markedly decreased both activities. However, heat denaturation destroyed their ionophorous ability but preserved their mitogenicity, thereby demonstrating that ionophorous activity was not essential for B cell activation. These data suggest that B cell mitogens do not necessarily act as primary ionophores. However, we propose that these molecules intercalate into the lipid portion of the cell membrane, and that this interaction initiates the process of B cell activation.