Article
Development of NK cells with chemotaxis for SDF-1alpha secreting gliomas
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Authors
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Nadja Müller
- Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum Carl Gustav Carus, TU Dresden, Dresden
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Gabriele Schackert
- Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum Carl Gustav Carus, TU Dresden, Dresden
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Achim Temme
- Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum Carl Gustav Carus, TU Dresden, Dresden
| Published: | May 21, 2013 |
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Outline
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Objective: Immunotherapeutic strategies for the treatment of gliomas have shown promising results in pre-clinical studies. However, immunotherapy of brain tumors is hampered by ineffective infiltration of immune effector cells at the tumor side. SDF-1alpha (SDF-1α) expression has been reported in gliomas. By transferring the CXCR4-chemokine receptor to primary NK cells we sought to guide these immune effector cells to SDF-1α secreting gliomas.
Method: U87-MG and U343-MG cells were genetically engineered to express soluble SDF-1α. The NK cell line YTS and primary NK cells were transduced with a lentiviral vector encoding for CXCR4. Surface expression of CXCR4 was assessed by FACS-staining. The capacity of NK cells to migrate towards an SDF-1α gradient was tested in transwell-assays. For in vivo testing SDF-1α-secreting U87 and the same number of EGFP-control cells were s.c. xenografted in Nu/Nu mice. Mice were either treated by four i.v. injections of red-labeled YTS-CXCR4 or YTS wt cells. After 8 days, mice were sacrificed and tumors were analyzed by fluorescence microscopy.
Results: We successfully generated U87-MG and U343-MG cell lines with high SDF-1α secretion and YTS-NK cells as well as primary human NK cells with expression of CXCR4. In-vitro migration-assays demonstrated a 2-fold to 3-fold increased migration of CXCR4-modified YTS NK-cells as well as primary NK cells towards SDF-1α-secreting glioma cells when compared to controls. Furthermore, this specific migration was completely blocked with the CXCR4 inhibitor AMD3100. Subcutaneous implantation of SDF-1α secreting glioma cells led to establishment of tumors, which even upon explantation after 21 days and passaging in cell culture were found to secrete high amounts of SDF-1α. After i.v. injection of YTS-NK cells we observed an improved invasion of YTS-CXCR4-positive cells into subcutaneous gliomas secreting SDF-1α when compared to SDF1α-negative tumors and when compared to CXCR4-negative YTS controls.
Conclusions: Augmenting effector cells of the immune system with specific migratory capacities towards a chemokine secreting tumor source promises to improve immunotherapeutic approaches of treatment of brain tumors and disseminated tumor cells.
