title: Potentiation of antiproliferative effects of monoclonal antibody Lym-1 and immunoconjugate Lym-1-gelonin on human Burkitt's lymphoma cells with gamma-interferon and tumor necrosis factor.
abstract: A type I ribosome inactivating protein, gelonin, was linked to Lym-1, a murine monoclonal antibody reactive with a polymorphic determinant of class II HLA-DR histocompatibility leukocyte antigen (HLA) on human lymphoma cells, via a disulfide linkage using the heterobifunctional cross-linking agent, N-succinimidyl-3-(2-pyridyldithio) propionate. This immunotoxin was purified from unreacted gelonin and unconjugated Lym-1 by fast protein liquid chromatography using sephacryl S-300 gel filtration and blue sepharose affinity gradient separation. Binding of Lym-1-gelonin immunoconjugate to human Raji Burkitt's lymphoma cells was demonstrated by indirect immunofluorescence using flow cytometry. Lym-1-gelonin was very active in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium salt and sulforhodamine B in vitro cytotoxicity assays against the Raji lymphoma cell line and confirmed the fact that monoclonal antibody Lym-1 internalizes into human lymphoma cells. A weaker cytostatic antiproliferative effect was also noted for unconjugated Lym-1. gamma-interferon augmented the antiproliferative effects of Lym-1-gelonin conjugate and unconjugated Lym-1, by having a direct cytotoxic effect on the Raji cells. Tumor necrosis factor-alpha also enhanced the antiproliferative effect of unconjugated Lym-1, but did not significantly augment the cytotoxic activity of the Lym-1-gelonin conjugate. These results suggest that anti-HLA class II monoclonal antibodies may be useful in constructing immunotoxins for the treatment of human lymphomas and leukemias expressing HLA class II antigens, and that unconjugated anti-HLA class II monoclonal antibodies may be therapeutically useful in conjunction with recombinant cytokines, especially gamma-interferon.
authors.affiliation: Department of Oncology, Montefiore Medical Center, Bronx, NY 10467, USA.
authors.forename: K P
authors.initials: KP
authors.lastname: O'Boyle
authors.forename: D
authors.initials: D
authors.lastname: Colletti
authors.forename: C
authors.initials: C
authors.lastname: Mazurek
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: S K
authors.initials: SK
authors.lastname: Ray
authors.forename: B
authors.initials: B
authors.lastname: Diamond
authors.forename: M G
authors.initials: MG
authors.lastname: Rosenblum
authors.forename: A L
authors.initials: AL
authors.lastname: Epstein
authors.forename: D
authors.initials: D
authors.lastname: Shochat
authors.forename: J P
authors.initials: JP
authors.lastname: Dutcher
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology*; D000972:Antineoplastic Agents, Phytogenic / Q000378:metabolism / Q000494:pharmacology*; D001666:Binding Sites, Antibody; D002051:Burkitt Lymphoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D002477:Cells; D004357:Drug Synergism; D006131:Growth Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D006801:Humans; D018796:Immunoconjugates / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology*; D051379:Mice; D010940:Plant Proteins / Q000378:metabolism / Q000494:pharmacology*; D054789:Ribosome Inactivating Proteins, Type 1; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D000972:Antineoplastic Agents, Phytogenic; D006131:Growth Inhibitors; D018796:Immunoconjugates; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D014409:Tumor Necrosis Factor-alpha; C025674:GEL protein, Gelonium multiflorum; D007371:Interferon-gamma