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The Journal of Immunology, 1979, 122: 723-727.
Copyright © 1979 by The American Association of Immunologists, Inc.

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Purification and Biochemical Characterization of Human Lymphocyte Mitogenic Factor (LMF)1

William D. Rutenberg2, Usama al-Khalidi3, Fred S. Rosen and Ezio Merler

Laboratory of Immunology, Department of Medicine, Children's Hospital Medical Center, and Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115

Abstract

Supernatants of human T lymphocytes stimulated by TT antigen release two factors that induce mitogenesis in autologous and allogeneic B lymphocytes. These factors are precipitated by 60% ammonium sulfate and 50% ethanol, and are both destroyed by heating to 70°C for 5 min. By equilibrium ultracentrifugation there was a peak of mitogenic activity in the fraction with a specific gravity of 1.3147 corresponding to a partial specific volume of 0.761. After ultrafiltration through an Amicon XM50 membrane, the concentrate was chromatographed on a Sephadex G-200 column. Mitogenic activity was found only in the post-albumin fraction. When the postalbumin fraction was run on an isoelectrofocusing column, two distinct mitogenic factors were identified. The major peak of mitogenic activity (LMF) had a pI of 6.68 ± 0.05 and the minor peak (MMF) had a pI of 7.27 ± 0.05. Amino acid analysis of LMF identified it as a protein and PAGE showed that LMF probably was a tetramer with a m.w. of 80,000.

Footnotes

1 This work was supported by United States Public Health Service Research Grant AI 05877 and United States Public Health Service Training Grants AI 00366 and HL 05855.

2 Present address: University of Illinois Medical Center, Chicago, Illinois.

3 Present address: American University of Beirut, Beirut, Lebanon.







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