Biochemical separation of a human B cell mitogenic factor
| Autor: | Richard J. Ford, Lawrence B. Lachman, Shashikant Mehta, John W. Morgan, Abby L. Maizel, Chintaman G. Sahasrabuddhe |
|---|---|
| Rok vydání: | 1982 |
| Předmět: |
DNA Replication
medicine.medical_treatment T cell Size-exclusion chromatography Cell Separation Biology Lymphocyte Activation Peripheral blood mononuclear cell medicine Humans Secretion Growth Substances Polyacrylamide gel electrophoresis Cells Cultured Ammonium sulfate precipitation B cell B-Lymphocytes Multidisciplinary Cell Cycle Molecular biology Kinetics Cytokine medicine.anatomical_structure Immunoglobulin M Interleukin-4 Research Article |
| Zdroj: | Proceedings of the National Academy of Sciences. 79:5998-6002 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.79.19.5998 |
| Popis: | Recent studies have established the ability of human B lymphocytes to undergo G1-phase cell cycle progression and subsequent DNA synthesis upon exposure to factor(s) present in media conditioned by lectin-stimulated mononuclear cells. Procedures for the isolation of such a cytokine have been the focus of the present investigation. Conditioned medium from cells stimulated by lectin for 72 hr was fractionated by ammonium sulfate precipitation, ion-exchange chromatography, and gel filtration chromatography. During the isolation procedure the proliferation-stimulating activity of the column fractions was assayed concurrently on purified human T cells, purified human B cells, and murine thymocytes. T cell and B cell stimulatory factors present in the initial conditioned medium were found to copurify during ammonium sulfate precipitation, DEAE-Sephadex chromatography, and Bio-Gel P-30 gel filtration. However, partial separation of these two activities was achieved after Bio-Gel P-100 gel filtration. Analytic polyacrylamide gel electrophoresis of radiolabeled Bio-Gel P-100 column fractions demonstrated a distinct protein band of 14,000-15,000 daltons in those column fractions predominantly supporting T cell growth and a distinct protein band of 12,000-13,000 daltons for those fractions predominantly supporting B cell growth. The fractions associated with B cell mitogenic activity induced B cell S-phase entry in a proportion of B lymphocytes in the absence of any detectable IgM secretion. |
| Databáze: | OpenAIRE |
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