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: K P O'Boyle, D Colletti, C Mazurek, Y Wang, S K Ray, B Diamond, M G Rosenblum, A L Epstein, D Shochat, J P Dutcher
• Journal: Journal of immunotherapy with emphasis on tumor immunology : official journal of the Society for Biological Therapy (J Immunother Emphasis Tumor Immunol) Vol. 18 Issue 4 Pg. 221-30 (Nov 1995) ISSN: 1067-5582 [Print] United States
• PMID: 8680650 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antibodies, Monoclonal
• Antineoplastic Agents, Phytogenic
• Growth Inhibitors
• Immunoconjugates
• Plant Proteins
• Ribosome Inactivating Proteins, Type 1
• Tumor Necrosis Factor-alpha
• GEL protein, Gelonium multiflorum
• Interferon-gamma

Topics

• Animals
• Antibodies, Monoclonal (metabolism, pharmacology)
• Antineoplastic Agents, Phytogenic (metabolism, pharmacology)
• Binding Sites, Antibody
• Burkitt Lymphoma (drug therapy, metabolism, pathology)
• Cells
• Drug Synergism
• Growth Inhibitors (pharmacology, toxicity)
• Humans
• Immunoconjugates (isolation & purification, metabolism, pharmacology)
• Interferon-gamma (pharmacology)
• Mice
• Plant Proteins (metabolism, pharmacology)
• Ribosome Inactivating Proteins, Type 1
• Tumor Cells, Cultured
• Tumor Necrosis Factor-alpha (pharmacology)