title: A functional recombinant human 4-1BB ligand for immune costimulatory therapy of cancer.
abstract: Costimulatory factors hold great promise for development into novel anticancer biotherapeutics. An agonist to 4-1BB is ranked number 8 by National Cancer Institute on the list of 20 agents with high potential for use in treating cancer. We earlier reported on a recombinant murine 4-1BB ligand fusion protein that binds 4-1BB receptor on murine T cells and stimulates their proliferation in tumor-bearing mice. To facilitate clinical translation,we constructed a corresponding recombinant human 4-1BB ligand fusion protein (hIg-h4-1BBLs) and showed its ability to activate human T cells in vitro. Using Chinese hamster ovary cells transformed with a plasmid coexpressing hIg-h4-1BBLs and rat glutamine synthetase, we generated a high-producing clone by sequential selection with methionine sulfoximine. The hIg-h4-1BBLs was partially purified by protein A column chromatography and characterized biochemically and functionally, using human 4-1BB binding and human T-cell proliferation assays, in vitro.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western Blot confirmed that the hIg-h4-1BBLs is expressed predominantly as a functionally active multimeric protein with the ability to specifically bind to cells expressing human 4-1BB receptor and induce significant T-cell proliferation in vitro using both human and monkey peripheral blood mononuclear cells. The hIg-h4-1BBLs can be produced in large quantities from the high producer clone and developed as a novel immune costimulatory biotherapeutic to treat, alone and in combination with other modalities, various malignant diseases in patients through T-cell activation. Process development of this clinical agent has been discussed with the Food and Drug Administration in a pre-Investigational New Drug meeting and presented to the Office of Biotechnology Activities in a public hearing.
authors.affiliation: Department of Gene and Cell Medicine, Mount Sinai School of Medicine, New York, NY 10029, USA.
authors.forename: Marcia
authors.initials: M
authors.lastname: Meseck
authors.forename: Tiangui
authors.initials: T
authors.lastname: Huang
authors.forename: Ge
authors.initials: G
authors.lastname: Ma
authors.forename: George
authors.initials: G
authors.lastname: Wang
authors.forename: Shu-Hsia
authors.initials: SH
authors.lastname: Chen
authors.forename: Savio L C
authors.initials: SL
authors.lastname: Woo
mesh_terms: D053330:4-1BB Ligand / Q000235:genetics* / Q000276:immunology*; D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D023061:Gene Order; D005974:Glutamate-Ammonia Ligase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000882:Haplorhini; D006801:Humans; D007167:Immunotherapy; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000628:therapy*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism
chemical_list: D053330:4-1BB Ligand; D000276:Adjuvants, Immunologic; D011993:Recombinant Fusion Proteins; D005974:Glutamate-Ammonia Ligase