Cell surface and cell cycle analysis of metal-induced murine T cell proliferation

The heavy metal cations Pb2+, Ni2+ and Zn2+ have previously been shown to induce T cell proliferation which required the presence of both T cells and Ia+ cells at the initiation of culture. This work has examined the ability of these metals to induce cell cycle entry as determined by acridine orange cell cycle analysis. Cell surface phenotype analysis, performed on splenocytes stimulated with optimum metal concentrations (100 microM), indicated that in vitro T cell recovery (growth and/or longevity) was enhanced by Pb2+, Ni2+, and Zn2+. Furthermore, simultaneous examination of cell surface phenotype and cell cycle progression (propidium iodide) indicated that the predominant cell type proliferating in response to these metals was Thy-1.2+. The metals differentially induced L3T4+ and Lyt-2+ cells to enter the cell cycle. The ability of various monoclonal antibodies to modulate metal-induced proliferation was examined. Anti-L3T4a, anti-I-A and anti-I-E blocked metal-induced proliferation. Anti-Lyt-2 only partially inhibited whereas anti-Lyt-1 was stimulatory. These results suggest that recognition of major histocompatibility complex-encoded class II molecules is required for the induction of proliferation by these metals (similar to the autologous mixed lymphocyte response).