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Matrix metalloproteinases produced by rat IL-2-activated NK cells
R P Kitson1, P M Appasamy, U Nannmark
1University of Pittsburgh Cancer Institute, Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, PA 15213, USA.
Journal of Immunology (Baltimore, Md. : 1950)
|May 9, 1998
Summary
Adoptively transferred IL-2-activated Natural Killer (A-NK) cells use matrix metalloproteinases (MMPs) to degrade extracellular matrices, aiding their accumulation in cancer metastases. MMPs are not essential for A-NK cell cytotoxicity.
Area of Science:
- Immunology
- Cancer Biology
- Cell Biology
Background:
- Adoptively transferred IL-2-activated Natural Killer (A-NK) cells have been observed to accumulate within cancer metastases.
- Previous electron microscopy studies showed A-NK cells binding to endothelial cells and traversing basement membranes in pulmonary metastases.
Purpose of the Study:
- To investigate the mechanisms by which A-NK cells accumulate within cancer metastases.
- To determine if A-NK cells produce matrix metalloproteinases (MMPs) and if these enzymes are involved in their migration and accumulation.
Main Methods:
- SDS-PAGE gelatin zymography to detect MMP activity in rat A-NK cells.
- Inhibition assays using BB-94 (MMP inhibitor) and 3,4-dichloroisocoumarin (serine protease inhibitor).
- Western blot analysis and reverse transcriptase-PCR to confirm MMP-2 and MMP-9 identity and presence.
- In vitro migration assays using a model basement membrane-like extracellular matrix.
Main Results:
- Rat A-NK cells produce MMP-2 and MMP-9, with activities inhibited by BB-94.
- MMP-2 identity confirmed by Western blot, MMP-9 presence confirmed by RT-PCR.
- A-NK cells demonstrated the ability to migrate through a model basement membrane.
- BB-94 partially inhibited A-NK cell migration through the model basement membrane but did not affect cytotoxicity.
Conclusions:
- A-NK cells utilize BB-94-inhibitable matrix metalloproteinases (MMPs) to degrade extracellular matrices.
- MMPs likely play a role in the accumulation of A-NK cells within cancer metastases.
- MMPs are not involved in the cytolytic function of A-NK cells.