Purification and Characterization of an Inhibitor (Soluble Tumor Necrosis Factor Receptor) for Tumor Necrosis Factor and Lymphotoxin Obtained from the Serum Ultrafiltrates of Human Cancer Patients
Serum ultrafiltrates (SUF) from human patients with different types of cancer contain a blocking factor (BF) that inhibits the cytolytic activity of human tumor necrosis factor α (TNF-α) in vitro. BF is a protein with a molecular mass of 28 kDa on reducing sodium dodecyl sulfate/polyacrylamide gel e...
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Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 87; no. 22; pp. 8781 - 8784 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Washington, DC
National Academy of Sciences of the United States of America
01.11.1990
National Acad Sciences |
Subjects | |
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Abstract | Serum ultrafiltrates (SUF) from human patients with different types of cancer contain a blocking factor (BF) that inhibits the cytolytic activity of human tumor necrosis factor α (TNF-α) in vitro. BF is a protein with a molecular mass of 28 kDa on reducing sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The active material was purified to homogeneity by a combination of affinity chromatography, PAGE, and high-pressure liquid chromatography. Amino acid sequence analysis revealed that BF is derived from the membrane TNF receptor. Purified BF blocks the lytic activity of recombinant human and mouse TNF-α and recombinant human lymphotoxin on murine L929 cells in vitro. However, BF inhibits the lytic activity of TNF-α more effectively than it does that of lymphotoxin. The BF also inhibits the necrotizing activity of recombinant human TNF-α when coinjected into established cutaneous Meth A tumors in BALB/c mice. The BF may have an important role in (i) the regulation and control of TNF-α and lymphotoxin activity in cancer patients, (ii) interaction between the tumor and the host antitumor mechanisms, and (iii) use of systemically administered TNF-α in clinical trials with human cancer patients. |
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AbstractList | Serum ultrafiltrates (SUF) from human patients with different types of cancer contain a blocking factor (BF) that inhibits the cytolytic activity of human tumor necrosis factor alpha (TNF-alpha) in vitro. BF is a protein with a molecular mass of 28 kDa on reducing sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The active material was purified to homogeneity by a combination of affinity chromatography, PAGE, and high-pressure liquid chromatography. Amino acid sequence analysis revealed that BF is derived from the membrane TNF receptor. Purified BF blocks the lytic activity of recombinant human and mouse TNF-alpha and recombinant human lymphotoxin on murine L929 cells in vitro. However, BF inhibits the lytic activity of TNF-alpha more effectively than it does that of lymphotoxin. The BF also inhibits the necrotizing activity of recombinant human TNF-alpha when coinjected into established cutaneous Meth A tumors in BALB/c mice. The BF may have an important role in (i) the regulation and control of TNF-alpha and lymphotoxin activity in cancer patients, (ii) interaction between the tumor and the host antitumor mechanisms, and (iii) use of systemically administered TNF-alpha in clinical trials with human cancer patients. Serum ultrafiltrates (SUF) from human patients with different types of cancer contain a blocking factor (BF) that inhibits the cytolytic activity of human tumor necrosis factor α (TNF-α) in vitro. BF is a protein with a molecular mass of 28 kDa on reducing sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The active material was purified to homogeneity by a combination of affinity chromatography, PAGE, and high-pressure liquid chromatography. Amino acid sequence analysis revealed that BF is derived from the membrane TNF receptor. Purified BF blocks the lytic activity of recombinant human and mouse TNF-α and recombinant human lymphotoxin on murine L929 cells in vitro. However, BF inhibits the lytic activity of TNF-α more effectively than it does that of lymphotoxin. The BF also inhibits the necrotizing activity of recombinant human TNF-α when coinjected into established cutaneous Meth A tumors in BALB/c mice. The BF may have an important role in (i) the regulation and control of TNF-α and lymphotoxin activity in cancer patients, (ii) interaction between the tumor and the host antitumor mechanisms, and (iii) use of systemically administered TNF-α in clinical trials with human cancer patients. Serum ultrafiltrates (SUF) from human patients with different types of cancer contain a blocking factor (BF) that inhibits the cytolytic activity of human tumor necrosis factor {alpha} (TNF-{alpha}) in vitro. BF is a protein with a molecular mass of 28kDa on reducing sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The active material was purified to homogeneity by a combination of affinity chromatography, PAGE, and high-pressure liquid chromatography. Amino acid sequence analysis revealed that BF is derived from the membrane TNF receptor. Purified BF blocks the lytic activity of recombinant human and mouse TNF-{alpha} and recombinant human lymphotoxin activity of TNF-{alpha} and recombinant human lymphotoxin on murine L929 cells in vitro. However, BF inhibits the lytic activity of TNF-{alpha} more effectively than it does that of lymphotoxin. The BF also inhibits the necrotizing activity of recombinant human TNF-{alpha} when coinjected into established cutaneous Meth A tumors in BALB/c mice. The BF may have an important role in (i) the regulation and control of TNF-{alpha} and lymphotoxin activity in cancer patients, (ii) interaction between the tumor and the host antitumor mechanisms, and (iii) use of systemically administered TNF-{alpha} in clinical trials with human cancer patients. Serum ultrafiltrates (SUF) from human patients with different types of cancer contain a blocking factor (BF) that inhibits the cytolytic activity of human tumor necrosis factor alpha (TNF- alpha ) in vitro. BF is a protein with a molecular mass of 28 kDa on reducing sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The active material was purified to homogeneity by a combination of affinity chromatography, PAGE, and high-pressure liquid chromatography. The BF may have an important role in (i) the regulation and control of TNG- alpha and lymphotoxin activity in cancer patients, (ii) interaction between the tumor and the host antitumor mechanisms, and (iii) use of systemically administered TNF- alpha in clinical trials with human cancer patients. |
Author | Hwang, Chenduen Lucci, Joseph A. Cappuccini, Fabio Edward W. B. Jeffes Gatanaga, Tetsuya Yamamoto, Robert S. Lentz, Rigdon Granger, Gale A. Tomich, John Kohr, William |
AuthorAffiliation | Department of Molecular Biology and Biochemistry, University of California, Irvine 92717 |
AuthorAffiliation_xml | – name: Department of Molecular Biology and Biochemistry, University of California, Irvine 92717 |
Author_xml | – sequence: 1 givenname: Tetsuya surname: Gatanaga fullname: Gatanaga, Tetsuya – sequence: 2 givenname: Chenduen surname: Hwang fullname: Hwang, Chenduen – sequence: 3 givenname: William surname: Kohr fullname: Kohr, William – sequence: 4 givenname: Fabio surname: Cappuccini fullname: Cappuccini, Fabio – sequence: 5 givenname: Joseph A. surname: Lucci fullname: Lucci, Joseph A. – sequence: 6 fullname: Edward W. B. Jeffes – sequence: 7 givenname: Rigdon surname: Lentz fullname: Lentz, Rigdon – sequence: 8 givenname: John surname: Tomich fullname: Tomich, John – sequence: 9 givenname: Robert S. surname: Yamamoto fullname: Yamamoto, Robert S. – sequence: 10 givenname: Gale A. surname: Granger fullname: Granger, Gale A. |
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SubjectTerms | 550201 - Biochemistry- Tracer Techniques AMINO ACID SEQUENCE Amino acids Analytical, structural and metabolic biochemistry Animals BASIC BIOLOGICAL SCIENCES Biological and medical sciences Cancer Cytokines Cytotoxicity, Immunologic - drug effects DISEASES ELECTROPHORESIS Fundamental and applied biological sciences. Psychology Gels Humans In Vitro Techniques Lymphotoxin-alpha - antagonists & inhibitors LYSIS MEMBRANE PROTEINS Mice Miscellaneous Molecular Sequence Data MOLECULAR STRUCTURE Molecular Weight NECROSIS NEOPLASMS Neoplasms - blood Neoplasms, Experimental - pathology ORGANIC COMPOUNDS PATHOLOGICAL CHANGES PATIENTS Peptide Fragments - blood Phagocytes PROTEINS PURIFICATION RECEPTORS Receptors, Cell Surface - isolation & purification Receptors, Tumor Necrosis Factor Tumor necrosis factor receptors Tumor Necrosis Factor-alpha - antagonists & inhibitors Tumor necrosis factors Tumors |
Title | Purification and Characterization of an Inhibitor (Soluble Tumor Necrosis Factor Receptor) for Tumor Necrosis Factor and Lymphotoxin Obtained from the Serum Ultrafiltrates of Human Cancer Patients |
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