Distribution, cellular localization, and therapeutic potential of the tumor-associated antigen Ku70/80 in glioblastoma multiforme
- 178 Downloads
Antibodies specifically targeting tumor-associated antigens have proved to be important tools in the treatment of human cancer. A desirable target antigen should be unique to tumor cells, abundantly expressed, and readily available for antibody binding. The Ku70/80 DNA-repair protein is expressed in the nucleus of most cells; it is, however, also present on the cell surface of tumor cell lines, and antibodies binding Ku70/80 at the cell surface were recently shown to internalize into tumor cells. To evaluate the potential of Ku70/80-antigen as a therapeutic target for immunotoxins in glioblastoma multiforme, we investigated binding and localization of Ku70/80-specific antibodies in tissue samples from glioblastomas and normal human brains, and in glioma cell cultures. Furthermore, the internalization and drug-delivery capacity were evaluated by use of immunotoxicity studies. We demonstrate that Ku70/80 is localized on the cell plasma membrane of glioma cell lines, and is specifically present in human glioblastoma tissue. Antibodies bound to the Ku70/80 antigen on the cell surface of glioma cells were found to internalize via endocytosis, and shown to efficiently deliver toxins into glioblastoma cells. The data further imply that different antibodies directed against Ku70/80 possess different abilities to target the antigen, in relation to its presentation on the cell surface or intracellular localization. We conclude that Ku70/80 antigen is uniquely presented on the plasma membrane in glioblastomas, and that antibodies specific against the antigen have the capacity to selectively bind, internalize, and deliver toxins into tumor cells. These results imply that Ku70/80 is a potential target for immunotherapy of glioblastoma multiforme.
KeywordsGlioma Tumor antigen Immunotherapy Ku70/80
Confocal laser-scanning microscopy
Differential interference contrast
Epidermal growth-factor receptor
We gratefully acknowledge Anna Ebbesson at the Department of Experimental Pathology and LBIC optical unit, Lund University, for technical support with immunofluorescence and immunohistochemistry; Susanne Strömblad and Catarina Blennow, Department of Neurosurgery and The Rausing Laboratory, Lund University, for technical support with cell culture and cryosectioning; Peter Ekström, Department of Cell and Organism Biology, Lund University, for the use of the Zeiss confocal microscope in some of the CLSM analyses; and the expert assistance of the Mammalian Protein Expression Core Facility at the University of Gothenburg. This investigation was supported by grants from The Hans and Märit Rausing Charitable Trust, and the Sten Lexner fund.
- 5.Bigner DD, Brown MT, Friedman AH, Coleman RE, Akabani G, Friedman HS, Thorstad WL, McLendon RE, Bigner SH, Zhao XG, Pegram CN, Wikstrand CJ et al (1998) Iodine-131-labeled antitenascin monoclonal antibody 81C6 treatment of patients with recurrent malignant gliomas: phase I trial results. J Clin Oncol 16:2202–2212PubMedGoogle Scholar
- 6.Sampson JH, Akabani G, Archer GE, Bigner DD, Berger MS, Friedman AH, Friedman HS, Herndon JE II, Kunwar S, Marcus S, McLendon RE, Paolino A et al (2003) Progress report of a phase I study of the intracerebral microinfusion of a recombinant chimeric protein composed of transforming growth factor (TGF)-alpha and a mutated form of the Pseudomonas exotoxin termed PE-38 (TP-38) for the treatment of malignant brain tumors. J Neurooncol 65:27–35CrossRefPubMedGoogle Scholar
- 17.Salford LG, Siesjo P, Skagerberg G, Persson BRR, Larsson E-M, Lindvall M, Visse E, Widegren B (2002) Search for effective therapy against glioblastoma multiforme—clinical immunisation with autologous glioma cells transduced with the human interferon-gamma gene. Int Congr Ser 1247:211–220CrossRefGoogle Scholar