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

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...

Full description

Saved in:
Bibliographic Details
Published in:Molecular immunology 1995-08, Vol.32 (12), p.853-864
Main Authors: Wagner, Ludwig, Avery, Robin K., Bensinger, Lisa, Kusinitz, Felicity, Hibberd, Patricia L., Pasternack, Mark S.
Format: Article
Language:English
Subjects:
Animals
apoptosis
Apoptosis - immunology
Apoptosis - physiology
aprotinin
Aprotinin - immunology
Aprotinin - physiology
Cell Line
Cell Membrane - immunology
Cell Membrane - physiology
cell-mediated cytotoxicity
cytotoxic T-lymphocyte
Cytotoxicity, Immunologic
granzyme A
Granzymes
Kinetics
Mice
Models, Biological
protease inhibition
Serine Endopeptidases - physiology
Serine Proteinase Inhibitors - immunology
Serine Proteinase Inhibitors - physiology
T-Lymphocytes, Cytotoxic - immunology
T-Lymphocytes, Cytotoxic - physiology
Tumor Cells, Cultured
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0161-5890
1872-9142
DOI:10.1016/0161-5890(95)00054-I