A single chain immunotoxin, targeting the melanoma-associated chondroitin sulfate proteoglycan, is a potent inducer of apoptosis in cultured human melanoma cells
| Autor: | Michael Schwenkert, Matthias Peipp, Jan Dörrie, Katrin Birkholz, Christian Kellner, Dirk M. Nettelbeck, Niels Schaft, Georg H. Fey, Beatrice Schuler-Thurner, Eckhart Kämpgen, Markus Kügler, Soldano Ferrone, Michael Schwemmlein |
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| Rok vydání: | 2008 |
| Předmět: |
Cytotoxicity
Immunologic Cancer Research Programmed cell death Virulence Factors Recombinant Fusion Proteins Bacterial Toxins Exotoxins Apoptosis Dermatology Biology Article chemistry.chemical_compound Immunotoxin Cell Line Tumor Cyclosporin a Tumor Cells Cultured Humans Cytotoxic T cell Pseudomonas exotoxin Melanoma ADP Ribose Transferases Immunotoxins Molecular biology Chondroitin Sulfate Proteoglycans Oncology chemistry Cell culture Chondroitin sulfate proteoglycan Cyclosporine Immunotherapy Immunosuppressive Agents |
| Zdroj: | Melanoma Research. 18:73-84 |
| ISSN: | 0960-8931 |
| Popis: | A recombinant immunotoxin was constructed by fusing a single chain fragment variable antibody fragment, specific for the melanoma-associated chondroitin sulfate proteoglycan (MCSP), to a truncated variant of Pseudomonas exotoxin A (ETA'), carrying a C-terminal KDEL-peptide for improved retrograde intracellular transport. The resulting immunotoxin MCSP-ETA' was periplasmatically expressed in Escherichia coli and purified under native conditions by affinity chromatography resulting in a yield of approximately 30 mug/l bacterial culture. This immunotoxin induced antigen-specific apoptosis in the cultured human melanoma-derived cell lines A2058 and A375M, and treatment with a single dose of the agent eliminated up to 80% of these cells within 72 h. The dose needed for half-maximum killing (EC50) was approximately 1 nmol/l for both cell lines. MCSP-ETA' also displayed cytotoxic activity against cultured primary melanoma cells from patients with advanced disease (pathologic stages IIIC and IV), with net cell death reaching up to 70% within 96 h after treatment with a single dose of 14 nmol/l. MCSP-ETA' induced cell death synergistically with cyclosporin A, both in established human melanoma cell lines and cultured primary melanoma cells. The distinctive antigen-restricted induction of apoptosis and the synergy with cyclosporin A justify further evaluation of this novel agent with regard to its potential application for the treatment of malignant melanoma. |
| Databáze: | OpenAIRE |
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