Chemical characterization of macrophage cytotoxicity factor, macrophage migration inhibitory factor, T-helper cell-replacing factor and colony-stimulating factor from culture supernatants of concanavalin A-stimulated murine spleen cells

• Published in: Immunobiology (1979) Vol. 157; no. 2; p. 169
• Main Authors: Hübner, L, Kniep, E M, Laukel, H, Sorg, C, Fischer, H, Gassel, W D, Havemann, K, Kickhöfen, B, Lohmann-Matthes, M L, Schimpl, A, Wecker, E
• Format: Journal Article
• Language: English
• Published: Netherlands 01.07.1980
• Subjects: Animals; Cells, Cultured; Chemical Fractionation; Chromatography, Gel; Colony-Stimulating Factors; Concanavalin A - pharmacology; Cytotoxicity, Immunologic; Isoelectric Focusing; Macrophage Migration-Inhibitory Factors; Macrophages - immunology; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Inbred DBA; Mice, Nude; Spleen - immunology; T-Lymphocytes - immunology
• ISSN: 0171-2985
• DOI: 10.1016/S0171-2985(80)80098-2

Supernatants from Concanavalin A-stimulated murine spleen cells were subjected to hydrophobic interaction chromatography on phenyl-Sepharose. Macrophage cytotoxicity factor (MCF), macrophage migration inhibitory factor (MIF), T-helper cell-replacing factor (TRF) and colony-stimulating factor (CSF) were bound at high ionic strength and were released stepwise at low ionic strength. CSF thus could be separated from MCF, MIF and TRF and the bulk of other proteins. Chromatograhy of pools containing MCF, MIF and TRF on Sephadex did not lead to a separation of the three activities which were all found in a molecular weight range of 25.000-55.000. Isoelectric focusing of these pools in pH range from 4 to 9 gave two peaks for MCF in a single sharp peak at pH 5.3. The results demonstrate that the four biological activities can be distinguished on a chemical basis and are accessible for purification and chemical characterization.