Lymphocyte Transformation Induced by Chemical Modification of Membrane Components

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Abstract
Reliable culture conditions and a valid 3H-thymidine labeling procedure were established for studies on the in vitro response of rat lymph node cells to treatment with the mitogen, sodium periodate. Six days after an initial periodate stimulation of rat lymph node cells (by which time all blasts have reverted to “secondary” lymphocytes and 3H-thymidine incorporation has declined to near background levels) the surviving cells were restimulated either directly, i.e., by retreatment of all the cells with periodate, or indirectly, i.e., in mixed cultures composed half of cells treated against with periodate, but blocked with mitomycin C (stimulator cells) and half of cells that received no retreatment (responder cells). Both the direct and indirect restimulation responses were faster and of greater magnitude than the corresponding initial responses. The magnitude of indirect restimulation was about 36% of that of direct restimulation. Reduction of the periodate-induced aldehyde groups with sodium borohydride abolished both the direct and indirect restimulation responses. The greater magnitude of the indirect restimulation response as compared with the initial indirect response was due to an enhanced ability of the secondary lymphocytes to respond rather than to an enhanced ability to stimulate. The responding cells in periodate-restimulated cultures were the progeny of the cells that had responded initially. Concanavalin A also restimulated at least some of the periodate progeny cells, and in addition stimulated some cells that had not responded to periodate initially. Cells from cultures initially stimulated by periodate were able to serve as indirect restimulators of cells initially stimulated by either periodate or concanavalin A. In contrast, cells initially stimulated by concanavalin A (and then treated with mitomycin C and periodate) could restimulate neither type of responder.

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