Inhibition of macrophage migration by a factor from ascites fluids of ovarian cancer patients. I. Biochemical characterization and purification.
A nonspecific inhibitor of macrophage migration was found in large quantities in the cell-free ascitic fluids from patients with ovarian tumours. MIF-like activity was assigned by the ability of various dilutions of ascitic fluids to inhibit migration of guinea pig macrophages from agarose droplets. The factor was purified in 3 subsequent steps including ion exchange chromatography, gel filtration, and isoelectric focusing. The data obtained indicate molecular heterogeneity according to net charge and molecular weights. The main MIF activity was found at about 45, 20, and 10 kD, respectively, and partially at less than 10 kD. Isoelectric focusing of the various MIF species revealed activity peaks in the pH range from 3.8 to 5.0. A further peak was detected at pH 6.0 in crude material. The factor was purified about 10 000-fold compared to the starting material. The action of OC-MIF was inhibited by L-fucose, and when target cells were incubated with alpha-L-fucosidase, they did not respond any longer to OC-MIF. Furthermore, purified MIF-like activity is a nondialyzable glycoprotein, sensitive to treatment with neuraminidase, chymotrypsin, trypsin and pronase, however, unaffected by incubation at 60 degrees C for 1 h. The physicochemical properties of OC-MIF activity studied are comparable to lymphocyte-derived conventional MIF.
D001201:Ascites; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005260:Female; D006801:Humans; D007525:Isoelectric Focusing; D008263:Macrophage Migration-Inhibitory Factors / Q000302:isolation & purification*; D009439:Neuraminidase / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000032:analysis*; D005644:alpha-L-Fucosidase / Q000378:metabolism
D008263:Macrophage Migration-Inhibitory Factors; D009439:Neuraminidase; D005644:alpha-L-Fucosidase