Effect of mitogenic and hormonal stimulation on zinc transport in mixed lymphocyte cultures

Human mixed lymphocyte cultures (hMLC) were used to examine the relationship between mitogenic stimuli (MITO), synthetic human calcitonin (hCT), and human insulin (hINS) on zinc (Zn) transport kinetics. Lymphocytes were isolated using a Ficoll density gradient. The hMLCs were labeled by incubation with 65Zn in either control or MITO-containing media. 65Zn release to equilibrium was then measured in unstimulated and mitogen-stimulated cells treated with hCT and hINS. hCT and hINS were added only during this final incubation due to the rapid response to peptide hormones. Bidirectional transmembrane flux coefficients were calculated using a closed two-compartment model. The hMLCs subjected to MITO stimulation demonstrated a 25% decrease in the fractional efflux coefficient (Kcm) and a 69% increase in the fractional influx coefficient (Kmc) compared with controls. Acute exposure to hINS resulted in a marked increase in Kmc with no significant change in Kcm. Acute exposure to hCT had effects qualitatively similar to those of MITO alone. Neither hormone significantly altered the transport of 65Zn when compared with stimulation with MITO alone.