Purification to homogeneity and partial characterization of cytotoxic lymphocyte maturation factor from human B-lymphoblastoid cells
| Autor: | D L Stremlo, A G Wolitzky, Jeffrey D. Hulmes, Frank John Podlaski, Alvin S. Stern, Philip C. Familletti, T Truitt, Richard Anthony Chizzonite, P M Quinn, Y C Pan |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Interleukin 2
T-Lymphocytes Lymphocyte Molecular Sequence Data Biology Lymphocyte Activation Chromatography Affinity Biological Factors medicine Humans Cytotoxic T cell Amino Acid Sequence Killer Cells Lymphokine-Activated Polyacrylamide gel electrophoresis Cells Cultured Chromatography High Pressure Liquid B-Lymphocytes Multidisciplinary Lymphokine-activated killer cell Lymphoblast Lymphokine Chromatography Ion Exchange Recombinant Proteins Kinetics medicine.anatomical_structure Biochemistry Cell culture Cytokines Interleukin-2 Research Article medicine.drug |
| Zdroj: | Proceedings of the National Academy of Sciences. 87:6808-6812 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.87.17.6808 |
| Popis: | A cytokine that can synergize with interleukin 2 to activate cytotoxic lymphocytes was purified to homogeneity. The protein, provisionally called cytotoxic lymphocyte maturation factor (CLMF), was isolated from a human B-lymphoblastoid cell line that was induced to secrete lymphokines by culture with phorbol ester and calcium ionophore. The purification method, utilizing classical and high-performance liquid chromatographic techniques, yielded protein with a specific activity of 8.5 x 10(7) units/mg in a T-cell growth factor assay. Analysis of the purified protein by sodium dodecyl sulfate/polyacrylamide gel electrophoresis demonstrated that CLMF is a 75-kDa heterodimer composed of disulfide-bonded 40-kDa and 35-kDa subunits. Determination of the N-terminal amino acid sequences of the two subunits revealed that both subunits are not related to any previously identified cytokine. Purified CLMF stimulated the proliferation of human phytohemagglutinin-activated lymphoblasts by itself and exerted additive effects when used in combination with suboptimal amounts of interleukin 2. Furthermore, the purified protein was shown to synergize with low concentrations of interleukin 2 in causing the induction of lymphokine-activated killer cells. |
| Databáze: | OpenAIRE |
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