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Characterisation of functional biotinylated TNF-alpha targeted to the membrane of apoptotic melanoma cells.
- Source :
-
Journal of immunological methods [J Immunol Methods] 2003 May 01; Vol. 276 (1-2), pp. 79-87. - Publication Year :
- 2003
-
Abstract
- The physiologic clearance of apoptotic cells prevents inflammation at the site of cell death and limits the immunogenicity of tumors. In this study we report the functional characterisation of biotinylated tumor necrosis factor-alpha (TNF-alpha) after anchorage to apoptotic melanoma cells via a biotin-avidin-biotin bridge. Flow cytometric and morphological analysis showed that biotinylated TNF-alpha efficiently bound to apoptotic membrane blebs of dying cells. Membrane-bound TNF-alpha (12 fg/cell) killed sensitive WEHI164 cells 250-fold more effectively than equivalent amounts of the soluble cytokine. Furthermore, macrophages engulfing apoptotic cells with membrane-bound TNF-alpha secreted significantly higher amounts of soluble TNF-alpha and lower amounts of interleukin-10 (IL-10). Therefore the bridging of TNF-alpha potentiates its biological function and influences the outcome of the phagocytic clearance of apoptotic tumor cells.
- Subjects :
- Animals
Biotinylation
Cell Membrane chemistry
Cell Membrane drug effects
Cytokines biosynthesis
Drug Delivery Systems
Flow Cytometry
Macrophages immunology
Melanoma, Experimental immunology
Melanoma, Experimental pathology
Mice
Phagocytosis
Tumor Cells, Cultured
Tumor Necrosis Factor-alpha analysis
Tumor Necrosis Factor-alpha chemistry
Apoptosis
Melanoma, Experimental drug therapy
Tumor Necrosis Factor-alpha therapeutic use
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1759
- Volume :
- 276
- Issue :
- 1-2
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 12738361
- Full Text :
- https://doi.org/10.1016/s0022-1759(03)00069-3