Related Experiment Video
Updated: Aug 12, 2026
Methods to Assess Beta Cell Death Mediated by Cytotoxic T Lymphocytes
Published on: June 16, 2011
Resistance of mouse cytolytic cells to pore-forming protein-mediated cytolysis
Y Shinkai1, H Ishikawa, M Hattori
1Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan.
Abstract:
Pore-forming protein (perforin, PFP) was isolated from a mouse large granular lymphocyte (LGL) [natural killer (NK-like)] cell line. Purified PFP lysed a variety of mouse tumor cell lines and helper T lymphocyte cell lines. However, LGL and cytotoxic T lymphocyte cell lines were resistant to PFP-mediated cell lysis. The presence of hemolytic activity in the granule was examined in these resistant cell lines. Four out of five of these resistant cell lines had hemolytically active granules. We determined whether NK cells freshly isolated from BALB/c nude mouse spleens were resistant to PFP-mediated cytolysis. Nylon column-passed spleen cells with an enriched content of NK cells exhibited more resistance than whole spleen cells. Moreover, when spleen cells were treated with PFP the remaining live cells showed enriched NK activity suggesting that normal peripheral cells with NK activity are resistant to PFP. These results indicate that cytolytic cells containing PFP have developed defense mechanisms to inhibit PEP-mediated cell lysis.
Insights
Cytolytic cells containing pore-forming protein (PFP) resist PFP-mediated lysis. Freshly isolated natural killer (NK) cells show resistance, indicating defense mechanisms against PFP.
Area of Science:
- Immunology
- Cell Biology
Background:
- Pore-forming protein (PFP) is crucial for cell-mediated cytotoxicity.
- Large granular lymphocyte (LGL) and natural killer (NK) cells utilize PFP for lysis.
Purpose of the Study:
- To investigate the resistance of LGL and NK cells to PFP-mediated lysis.
- To explore potential defense mechanisms in cytolytic cells.
Main Methods:
- Isolation of PFP from a mouse LGL cell line.
- Testing PFP-mediated lysis on various cell lines (tumor, T lymphocyte, LGL, CTL).
- Assessing hemolytic activity in resistant cell lines.
- Evaluating resistance of freshly isolated mouse NK cells to PFP.
Main Results:
- LGL and cytotoxic T lymphocyte (CTL) cell lines were resistant to PFP lysis.
- Resistant cell lines often possessed hemolytically active granules.
- Freshly isolated NK cells exhibited resistance to PFP-mediated cytolysis.
- NK cell activity was enriched in cells surviving PFP treatment.
Conclusions:
- Cytolytic cells, including NK cells, possess intrinsic defense mechanisms against PFP.
- These defense mechanisms inhibit self-lysis by PFP.
- Understanding these mechanisms is key to PFP-based immunotherapies.
