In vitro efficacy of recombinant diphtheria toxin-murine interleukin-4 immunoconjugate on mouse glioblastoma and neuroblastoma cell lines and the additive effect of radiation.
OBJECTIVE
The prognosis for patients with primary malignant brain tumors is poor despite aggressive treatment, and tumor recurrence is common regardless of the chosen therapy. Although multimodal treatment does not provide a cure, it is necessary to determine which treatment modalities have the greatest cytotoxic effect and can potentially prolong survival. Immunotoxin therapy is a novel approach for the treatment of tumors, and it has been successfully used in the central nervous system. Because the interleukin (IL)-4 receptor is commonly expressed on brain tumor cells, the purpose of this study was to evaluate the cytotoxic effect of using a modified diphtheria toxin-murine IL-4 (DT390- mIL4) immunoconjugate for the treatment of murine brain tumor cell lines and to determine whether the addition of radiation therapy could potentiate the effect of this agent.


METHODS
Spontaneous murine glioblastoma (SMA-560) and two neuroblastoma (Neuro-2a and NB41A3) cell lines were treated using DT390-mIL4 at different concentrations, and the anti-mouse IL-4 monoclonal antibody (11B11) was used for blocking its cytotoxicity. Other SMA-560 and Neuro-2a cell lines were treated using 500 cGy of radiation 3 hours before DT390-mIL4 treatment. Cytotoxity was evaluated using a trypan blue viability assay. The immunoconjugate exhibited a dose-dependent cytotoxic effect with 50% inhibitory concentration values of 0.56 x 10(-9) M in SMA-560, 1.28 x 10(-9) M in Neuro-2a, and 0.95 x (-10) M in NB41A3 cell lines. The cytotoxicity of DT390-mIL4 was specifically blocked by an excess of 11B11. Cytotoxicity was additive when the DT390-mIL4 at 10(-9) M immunoconjugate administration was followed by radiation therapy.


CONCLUSIONS
These results indicate that the IL-4 receptor can be a target for diphtheria toxin fusion proteins and that radiation can potentiate the effects of DT390-mIL4. The development of multimodal approaches to treat malignant brain tumors with agents that have different mechanisms of action may be beneficial.
Kim
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Department of Neurolosurgery, University of Minnesota School of Medicine, Minneapolis, Minnesota, USA.
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D000818:Animals; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000532:radiotherapy*; D045744:Cell Line, Tumor; D003131:Combined Modality Therapy; D003603:Cytotoxins / Q000276:immunology / Q000494:pharmacology; D004167:Diphtheria Toxin / Q000276:immunology / Q000494:pharmacology; D005909:Glioblastoma / Q000188:drug therapy* / Q000532:radiotherapy*; D018796:Immunoconjugates / Q000276:immunology / Q000494:pharmacology; D007167:Immunotherapy / Q000379:methods; D066298:In Vitro Techniques; D015847:Interleukin-4 / Q000276:immunology / Q000494:pharmacology*; D051379:Mice; D009447:Neuroblastoma / Q000188:drug therapy / Q000532:radiotherapy; D019948:Receptors, Interleukin-4 / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology
D003603:Cytotoxins; D004167:Diphtheria Toxin; D018796:Immunoconjugates; D019948:Receptors, Interleukin-4; D011994:Recombinant Proteins; D015847:Interleukin-4