Macrophage Migration Inhibitory Factor (MIF) Produced by a Human T Cell Hybridoma Clone

J-STAGE
  • HIROSE Shin-ichiro
    Biosciences Research Laboratories, Sankyo Co., Ltd.
  • OOKI Shinji
    Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical Sciences, The University of Tokyo
  • HIGUCHI Masahiro
    Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical Sciences, The University of Tokyo
  • OSAWA Toshiaki
    Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical Sciences, The University of Tokyo

抄録

A human T cell hybridoma clone, F5, producing high levels of macrophage migration inhibitory factor (MIF) was established by the emetine-actinomycin D selection method. This clone produced two species of MIF which were separated on a Phenyl Sepharose column. We purified MIF-2 (the more hydrophobic species of the two) to homogeneity from the conditioned medium of stimulated F5 cells by a series of steps that included hydrophobic chromatography, ion-exchange chromatography, Ricinus communis lectin affinity chromatography, and high-performance liquid chromatography on anion exchange and reverse-phase columns. Purified MIF was digested with endoproteinase Lys-C and Asp-N. The amino acid sequences of the generated peptides were determined. No sequence similarity with any other protein was found. The molecular weight of MIF-2 was estimated to be 45kDa from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of immunoprecipitates with anti-peptide antibodies. These results show that F5MIF-2 is a novel cytokine.

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