Lymphocyte transformation induced by chemical modification of membrane components. I. Characteristics of the direct and indirect restimulation responses of rat lymph node cells to periodate

Beyer, C.F.; Bowers, W.E.

Journal of Immunology 119(6): 2120-2128

1977


ISSN/ISBN: 0022-1767
PMID: 199671
Accession: 068523790

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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 directly, (i.e., by retreatment of all the cells with periodate) or indirectly [i.e., in mixed cultures composed half of cells treated again with periodate, but blocked with mitomycin C (stimulator cells) and half of cells that received no retreatment (responder cells)]. 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 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 no responded to periodate initially. Cells from cultures initially stimulated by periodate were able to serve as indirect restimulators of cells initially stimulated by periodate or concanavalin A. Cells initially stimulated by concanavalin A (and then treated with mitomycin C and periodate) could restimulate neither type of responder. The progeny cells from periodate stimulation differ from fresh cells in the kinetics and magnitude of the proliferative responses and in their ability to particulate in the cellular interactions required for indirect stimulation. Some of the changes in indirect periodate responses recall similar differences between the primary and secondary responses in the mixed lymphocyte reaction.