Density Gradient Electrophoresis of Mouse Spleen Lymphocytes

Abstract

Preparative density gradient electrophoresis has been employed for the separation of BALB/c mouse spleen T and B lymphocytes, on the basis of their surface charge. The high mobility cells were found to be predominantly T lymphocytes, whereas the low mobility cells were B lymphocytes. One way mixed lymphocyte cultures were prepared using electrophoretically separated BALB/c lymphocytes, either as responding or stimulating cells and unfractionated CBA/H/T6j mouse spleen lymphocytes, in the appropriate combination. The responding lymphocytes were found only among the high mobility cells which are primarily T lymphocytes. In contrast, the stimulating cells were found in the intermediate and low mobility fractions which contain the B lymphocytes and macrophages. Minimal stimulation in mixed lymphocyte culture was observed by T cells in this allogeneic system. Ia-positive cells were found only in the intermediate and low mobility fractions, which exhibited the capacity to stimulate allogeneic cells in MLC. Macrophages, identified by nonspecific esterase staining, exhibited intermediate electrophoretic mobility, their peak height coinciding with the maximum observed stimulation.