Abstract
Some of extracellular serine proteases with trypsin-like specificity of cleavage have been known to increase the release of inflammatory mediators from various cell types. For instance, two well-known trypsin-like serine proteases circulating in blood, granzyme A (GrA) and thrombin, have been found to promote interleukin (IL)-8 release from an alveolar epithelial A549 cell line. However, the mechanisms by which the proteases promote IL-8 release from the cells are not fully understood. In the present study, using A549 cells we found that (1) thrombin promoted IL-8 release from the cells via a mechanism partially involving activation of protease-activated receptor-1, a G-protein coupled receptor, whereas a recombinant form of GrA (rGrA) did it via a mechanism that does not involve the receptor activation; that (2) unlike rGrA, thrombin did not cause detachment and microtubule disruption of the cells; and that (3) the release of IL-8 induced by rGrA was inhibited in the presence of taxol, a microtubule-stabilizing reagent, whereas that induced by thrombin was not. These findings suggest that rGrA and thrombin promote the release of IL-8 from A549 cells through distinct mechanisms.
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Abbreviations
- BLT:
-
N-α-benzyloxy l-lysine thiobenzyl ester
- DTNB:
-
5,5′-Dithiobis-(2-nitrobenzoic acid)
- GrA:
-
Granzyme A
- IL:
-
Interleukin
- PARs:
-
Protease-activated receptors
- rGrA:
-
Recombinant form of rat GrA
- SFM:
-
Serum-free Dulbecco’s modified Eagle medium supplemented with 0.1% w/v BSA
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Acknowledgments
This work was supported by Grants-in-Aid for Scientific Research (18580118 to S. Tsuzuki) from the Japan Society for the Promotion of Science.
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Yoshikawa, Y., Hirayasu, H., Tsuzuki, S. et al. Granzyme A and thrombin differentially promote the release of interleukin-8 from alveolar epithelial A549 cells. Cytotechnology 62, 325–332 (2010). https://doi.org/10.1007/s10616-010-9275-z
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DOI: https://doi.org/10.1007/s10616-010-9275-z