Characterisation of functional biotinylated TNF-α targeted to the membrane of apoptotic melanoma cells

Valérie S. Zimmermann, Attilio Bondanza, Patrizia Rovere-Querini, Barbara Colombo, Angelina Sacchi, Umberto Fascio, Angelo Corti, Angelo A. Manfredi

Research output: Contribution to journalArticlepeer-review


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-α (TNF-α) after anchorage to apoptotic melanoma cells via a biotin-avidin-biotin bridge. Flow cytometric and morphological analysis showed that biotinylated TNF-α efficiently bound to apoptotic membrane blebs of dying cells. Membrane-bound TNF-α (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-α secreted significantly higher amounts of soluble TNF-α and lower amounts of interleukin-10 (IL-10). Therefore the bridging of TNF-α potentiates its biological function and influences the outcome of the phagocytic clearance of apoptotic tumor cells.

Original languageEnglish
Pages (from-to)79-87
Number of pages9
JournalJournal of Immunological Methods
Issue number1-2
Publication statusPublished - May 1 2003


  • Cell death
  • Immunotherapy
  • Melanoma
  • TNF-α

ASJC Scopus subject areas

  • Biotechnology
  • Immunology


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