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 supematants 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 timulated 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.

Murine macrophage-lymphocyte interactions: scanning electron microscopic study

“…The increase in macrophage spreading due to the MIF-containing fractions is similar to previously reported results (42,57,58). In addition, it appears as though this more rapid spreading occurs in the presence of spleen, thymic, or peritoneal-cavity lymphocytes and leads to an earlier attachment of the lymphocytes to the macrophages.…”

“…MIF. MIF was prepared as described previously (58). Briefly, spleen cell suspensions from either Brucella-infected or uninfected female Swiss-Webster ROR mice were incubated in medium NCTC-109 with B. abortus antigen at 37°C in a 5% C02-95% air atmosphere for 24 h. The supernatant fluid was collected, centrifuged to remove cells and cell debris, filtered through a microporous filter (0.45-am approximate pore diameter), and concentrated five times by way of ultrafiltration (Millipore Pellicon filter; nominal molecular weight limit, 1,000).…”