Inhibition of cytotoxic T-lymphocyte-triggered apoptosis by target cell surface-coupled aprotinin

Ludwig Wagner, Robin K. Avery, Lisa Bensinger, Felicity Kusinitz, Patricia L. Hibberd, Mark S. Pasternack

Research output: Contribution to journalArticlepeer-review

Abstract

A variety of recent investigations have implicated granzymes A and/or B in the target cell nuclear injury which accompanies cytotoxic T-lymphocyte-mediated cytolysis. Since soluble antiproteases have has limited efficacy in inhibiting CTL-mediated lysis, we developed a method to couple aprotinin, a peptide inhibitor of serine proteases, to the surface of target cells. Aprotinin modified by N-succinimidyl 3-(2-pyridyldithio)propionate retained trypsin-inhibitory activity, and target cells modified with aprotinin had demonstrable cell surface trypsin-inhibitory activity. Flow cytometry demonstrated that aprotinin was detectable on the target cell surface but underwent modulation at a rather rapid rate. When radiolabeled, aprotinin-coupled target cells were studied in 1-2 hr CTL assays, 51Cr release was little affected, but 125IUdR release was reduced up to 75% compared to controls. Corresponding apoptosis analysed by agarose gel electrophoresis and direct cytologic visualization was similarly reduced. Thus, aprotinin bound to the surface of target cells selectively protected target cells against CTL-mediated nuclear injury, and may serve as a model for the development of novel inhibitors of CTL-mediated lysis.

Original languageEnglish (US)
Pages (from-to)853-864
Number of pages12
JournalMolecular Immunology
Volume32
Issue number12
DOIs
StatePublished - Aug 1995
Externally publishedYes

Keywords

  • apoptosis
  • aprotinin
  • cell-mediated cytotoxicity
  • cytotoxic T-lymphocyte
  • granzyme A
  • protease inhibition

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

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