Granzyme B is a novel interleukin-18 converting enzyme

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• 作者：Youichi Omoto ; Keiichi Yamanaka ; Kazuya Tokime ; Shigehisa Kitano ; Masato Kakeda ; Tomoko Akeda ; Ichiro Kurokawa ; Esteban C. Gabazza ; Hiroko Tsutsui ; Naoyuki Katayama ; Kiyofumi Yamanishi ; Kenji Nakanish
• 刊名：Journal of Dermatological Science
• 出版年：2010
• 出版时间：August 2010
• 年：2010
• 卷：59
• 期：2
• 页码：129-135
• 全文大小：370 K

文摘

Background

Granzyme B (GrB) is recognized to induce apoptosis; however, little is known about its possible role in other biological events. IL-18, a potent inflammatory cytokine, is produced as an inactive precursor (proIL-18). Several cells, including monocytes/macrophage lineage and non-hematopoietic cells such as keratinocytes, produce proIL-18. ProIL-18 requires appropriate processing to become active. Caspase-1 is the authentic IL-18 processing enzyme and is essential for IL-18 release from monocyte/macrophage lineage cells. However, caspase-1 is absent in non-hematopoietic cells, suggesting that there is another candidate to cleave proIL-18 except for caspase-1.

Objective

GrB can invade and be active in cytoplasm of non-hematopoietic cells via perforin, therefore we investigated whether GrB converts proIL-18 into the biologically active form.

Methods

Recombinant proIL-18 (rproIL-18) was produced and purified for protease reaction with GrB; this incubate was evaluated by immunoblotting. Biological activity of the proteolytic fragment cleaved by GrB was determined by IFN-γ assay using KG-1 cells. IFN-γ induction was also analyzed between extracts from GrB(+)/caspase-1(−) human CD8+ T cells and proIL-18 from normal human keratinocytes (NHK).

Results

The proteolytic fragment that GrB cleaved proIL-18 had the same sequence and biological activity compared with mature IL-18 cleaved by caspase-1. Culture extracts from CD8+ T cells was able to cleave proIL-18 into authentic mature IL-18. IFN-γ induction was also detected in NHK treated with CD8+ T cells.

Conclusion

GrB is a potent IL-18 converting enzyme and suggest that GrB secreted by CTLs and/or NK cells may initiate IL-18 release from target cells, leading to the development of inflammation.