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

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 87; no. 22; pp. 8781 - 8784
• Main Authors: Gatanaga, Tetsuya, Hwang, Chenduen, Kohr, William, Cappuccini, Fabio, Lucci, Joseph A., Edward W. B. Jeffes, Lentz, Rigdon, Tomich, John, Yamamoto, Robert S., Granger, Gale A.
• 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: 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; Proteins; Cytotoxicity; Inhibition
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.87.22.8781

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.