Macrophage migration inhibitory factor (MIF) produced by a human T cell hybridoma clone.
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 45 kDa from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of immunoprecipitates with anti-peptide antibodies. These results show that F5MIF-2 is a novel cytokine.
Hirose
S
S
Biosciences Research Laboratories, Sankyo Co., Ltd., Tokyo.
Ooki
S
S
Higuchi
M
M
Osawa
T
T
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D002999:Clone Cells; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006825:Hybridomas; D008263:Macrophage Migration-Inhibitory Factors / Q000096:biosynthesis* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D013601:T-Lymphocytes / Q000276:immunology*
D008263:Macrophage Migration-Inhibitory Factors