The aim of this study was to construct a fusion protein from the cytokine granulocyte/macrophage colony-stimulating factor (GM-CSF) and a single-chain Fv fragment (scFv D29) and to investigate its potential to activate cells of the immune system against neuroblastoma cells expressing neural cell adhesion molecule (NCAM). Mammalian cell expression of the scFv D29-GM-CSF fusion protein was compared using a number of vectors, including retroviral and adenoviral vectors. The resultant fusion protein, expressed by HeLa cells, was found by ELISA to bind immobilized recombinant NCAM. Moreover, FACS analysis confirmed binding to the human neuroblastoma cell line SKNBE and a murine neuroblastoma cell line engineered to express the glycosylphosphatidylinositol form of human NCAM (N2A-rKNIE). The fusion protein was also found to stimulate the proliferation of the FDC-P1 haemopoietic cell line, which is dependent on GM-CSF (or interleukin 3) for continued growth. In vitro clonogenic assays indicated that scFv-GM-CSF could selectively induce growth inhibition of SKNBE cells by murine lymphoid cells.
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Conference Article|
August 01 2002
Targeted cytokine delivery to neuroblastoma
P. K. Dehal;
P. K. Dehal
1
1Medical Oncology, Paterson Institute for Cancer Research, Wilmslow Road, Withington, Manchester M20 4BX, U.K.
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M. J. Embleton;
M. J. Embleton
1Medical Oncology, Paterson Institute for Cancer Research, Wilmslow Road, Withington, Manchester M20 4BX, U.K.
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J. T. Kemshead;
J. T. Kemshead
1Medical Oncology, Paterson Institute for Cancer Research, Wilmslow Road, Withington, Manchester M20 4BX, U.K.
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R. E. Hawkins
R. E. Hawkins
2
1Medical Oncology, Paterson Institute for Cancer Research, Wilmslow Road, Withington, Manchester M20 4BX, U.K.
2To whom correspondence should be addressed (e-mail RHawkins@picr.man.ac.uk)
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Publisher: Portland Press Ltd
Received:
April 10 2002
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2002 Biochemical Society
2002
Biochem Soc Trans (2002) 30 (4): 518–520.
Article history
Received:
April 10 2002
Citation
P. K. Dehal, M. J. Embleton, J. T. Kemshead, R. E. Hawkins; Targeted cytokine delivery to neuroblastoma. Biochem Soc Trans 1 August 2002; 30 (4): 518–520. doi: https://doi.org/10.1042/bst0300518
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