1. Matrix metalloproteinases produced by rat IL-2-activated NK cells.
- Author
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Kitson RP, Appasamy PM, Nannmark U, Albertsson P, Gabauer MK, and Goldfarb RH
- Subjects
- Animals, Cell Movement immunology, Cells, Cultured, Collagenases immunology, Gelatinases immunology, Humans, Killer Cells, Natural cytology, Killer Cells, Natural enzymology, Male, Matrix Metalloproteinase 2, Matrix Metalloproteinase 9, Metalloendopeptidases immunology, RNA, Messenger analysis, Rats, Rats, Inbred F344, Collagenases biosynthesis, Gelatinases biosynthesis, Interleukin-2 pharmacology, Killer Cells, Natural immunology, Lymphocyte Activation drug effects, Metalloendopeptidases biosynthesis
- Abstract
We have previously documented that adoptively transferred IL-2-activated NK (A-NK) cells can accumulate within cancer metastases. Electron microscopic studies of pulmonary metastases have revealed that adoptively transferred A-NK cells that accumulate within metastases bind to endothelial cells and are able to traverse basement membranes. We have now extended these morphologic studies. We report that rat A-NK cells produce two matrix metalloproteinases: MMP-2 and MMP-9, as determined by SDS-PAGE gelatin zymography. These activities are inhibited following incubation with BB-94 (batimastat), a specific inhibitor of matrix metalloproteinases but not with 3,4-dichloroisocoumarin, an inhibitor of neutral serine proteases. The identity of MMP-2 was confirmed by Western blots using a polyclonal Ab against human MMP-2, whereas reverse transcriptase-PCR analysis of mRNA extracts of A-NK cells has confirmed the presence of MMP-9. In addition, we report for the first time that A-NK cells can migrate through a model basement membrane-like extracellular matrix. Moreover, the ability of A-NK cells to migrate through this model basement membrane was partially inhibited by BB-94; however, BB-94 has no effect on A-NK cell-mediated cytotoxicity, suggesting that matrix metalloproteinases do not contribute to cytolytic function of A-NK cells. In sum, our studies show that A-NK cells employ BB-94-inhibitable matrix metalloproteinases to degrade extracellular matrices. This suggests that matrix metalloproteinases may play a role in the accumulation of A-NK cells within cancer metastases.
- Published
- 1998