Cytoplasts from cytotoxic t lymphocytes are resistant to perforin-mediated lysis

David M. Ojcius*, Jiang Shibo, Pedro M. Persechini, Patricia A. Detmers, Ding-E Young John Ding-E Young

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

17 Scopus citations

Abstract

Cytotoxic T lymphocytes (CTL) contain a potent cytolytic pore-forming protein (PFP, perforin or cytolysin) localized in their cytoplasmic granules. In the presence of calcium, perforin lyses a variety of target cells (TC) non-specifically. CTL, however, are generally resistant to the lytic effect of perforin. In this work, cytoplasts from CTL and susceptible TC were made by centrifuging cells on a Ficoll density gradient in the presence of cytochalasin B. Characterization by electron microscopy and a serine esterase assay established that both CTL and TC cytoplasts were completely devoid of nuclei and CTL cytoplasts contained essentially no granules. CTL cytoplasts are just as resistant to perforin-mediated lysis as the intact CTL, and both TC and their corresponding cytoplasts are very sensitive to lysis. Furthermore, CTL cytoplasts are less effective than TC cytoplasts in inhibiting hemolysis, a property shared by the respective intact cells. These results indicate that soluble granular components do not confer resistance on CTL, and suggest that the protective agent(s) acts by impeding perforin binding to the CTL membrane.

Original languageEnglish
Pages (from-to)1011-1018
Number of pages8
JournalMolecular Immunology
Volume28
Issue number9
DOIs
StatePublished - 09 1991
Externally publishedYes

Keywords

  • Animal
  • Centrifugation, Density Gradient
  • Esterases
  • In Vitro
  • Membrane Proteins
  • Mice
  • Microscopy, Electron
  • Support, Non-U.S. Gov't
  • Support, U.S. Gov't, P.H.S.
  • T-Lymphocytes, Cytotoxic

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