Alterations in mouse macrophage migration: a function of assay systems, lymphocyte activation product preparation, and fractionation

Abstract

Supernatants from mouse spleen cell and peritoneal cell cultures were tested for the presence of lymphocyte activation products. Supernatants from mouse spleen cell and peritoneal cell cultures incubated with brucella antigens contained a macrophage migration inhibition factor(s) and a macrophage spreading factor(s) only if the cells were harvested from Brucella infected mice. After dialysis and freeze drying, the supernatants were fractionated by preparative acrylamide gel electrophoresis. Three fractions with lymphocyte activation product activity were obtained from the fractionated supernatants of mouse spleen cells and peritoneal cells harvested from Brucella infected mice and cultured with brucella antigen. One fraction inhibited mouse macrophage migration from capillary tubes but not from agarose wells. A second fraction not only inhibited macrophage migration from both agarose wells and capillary tubes, but also contained an activity(s) that stimulated macrophage migration through Nuclepore filters and induced macrophage spreading. A third fraction stimulated macrophage migration from agarose wells and also contained an activity(s) that stimulated macrophage migration through Nuclepore filters. Fractionated supernatants of mouse spleen cells and peritoneal cells harvested from uninfected mice incubated with and without brucella antigen, as well as of cells harvested from infected mice and not incubated with antigen, did not contain detectable lymphocyte activation products.