Induction of keratinocyte migration via transactivation of the epidermal growth factor receptor by the antimicrobial peptide LL-37
Induction of keratinocyte migration via transactivation of the epidermal growth factor receptor by the antimicrobial peptide LL-37
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DOI：
10.4049/jimmunol.175.7.4662
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发表时间：
2005-10-01
影响因子：
4.4
通讯作者：
Hashimoto, K
中科院分区：
文献类型：
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作者：
Tokumaru, S;Sayama, K;Hashimoto, K
皮肤伤口的闭合对于抵抗微生物病原体是必不可少的，并且角质形成细胞迁移是皮肤伤口愈合的重要步骤。Cathelicidin hCAP 18/LL-37是一种先天性抗菌肽，在皮肤中表达，用于消除微生物病原体。由于hCAP 18/LL-37在皮肤伤口部位上调，我们假设LL-37诱导角质形成细胞迁移。在这项研究中，我们发现1 μ g/ml LL-37在Boyden小室测定中诱导最大水平的角质形成细胞迁移。此外，LL-37在10分钟后磷酸化表皮生长因子受体（EGFR），这表明LL-37诱导的角质形成细胞迁移通过EGFR反式激活发生。为了验证这一假设，我们使用了阻断EGFR反式激活顺序步骤的抑制剂，如OSU 8 -1、CRM 197、抗EGFR 225号抗体和AG 1478。所有这些抑制剂完全阻断LL-37诱导的角质形成细胞迁移，这表明迁移通过HB-EGF介导的EGFR反式激活发生。此外，CRM 197、抗EGFR 225号和AG 1478阻断LL-37诱导的STAT 3磷酸化，并且用STAT 3的显性负突变体转染消除了LL-37诱导的角质形成细胞迁移，表明EGFR反式激活下游的STAT 3途径的参与。最后，我们测试了STAT 3的负调节因子的细胞因子信号传导抑制因子（SOCS）/含丝氨酸诱导的Src同源性2蛋白（CIS）家族是否调节LL-37诱导的角质形成细胞迁移。用SOCS 1/Jak 2结合蛋白或SOCS 3/CIS 3转染几乎完全消除LL-37诱导的角质形成细胞迁移。总之，LL-37通过肝素结合EGF介导的EGFR反式激活诱导角质形成细胞迁移，SOCS 1/Jak 2结合和SOCS 3/CIS 3负性调节这种迁移。该研究的结果表明，LL-37通过诱导角质形成细胞迁移来闭合皮肤伤口。
The closure of skin wounds is essential for resistance against microbial pathogens, and keratinocyte migration is an important step in skin wound healing. Cathelicidin hCAP18/LL-37 is an innate antimicrobial peptide that is expressed in the skin and acts to eliminate microbial pathogens. Because hCAP18/LL-37 is up-regulated at skin wound sites, we hypothesized that LL-37 induces keratinocyte migration. In this study, we found that 1 mu g/ml LL-37 induced the maximum level of keratinocyte migration in the Boyden chamber assay. In addition, LL-37 phosphorylated the epidermal growth factor receptor (EGFR) after 10 min, which suggests that LL-37-induced keratinocyte migration occurs via EGFR transactivation. To test this assumption, we used inhibitors that block the sequential steps of EGFR transactivation, such as OSU8-1, CRM197, anti-EGFR no. 225 Ab, and AG1478. All of these inhibitors completely blocked LL-37-induced keratinocyte migration, which indicates that migration occurs via HB-EGF-mediated EGFR transactivation. Furthermore, CRM197, anti-EGFR no. 225, and AG1478 blocked the LL-37-induced phosphorylation of STAT3, and transfection with a dominant-negative mutant of STAT3 abolished LL-37-induced keratinocyte migration, indicating the involvement of the STAT3 pathway downstream of EGFR transactivation. Finally, we tested whether the suppressor of cytokine signaling (SOCS)/cytokine-inducible Src homology 2-containing protein (CIS) family of negative regulators of STAT3 regulates LL-37-induced keratinocyte migration. Transfection with SOCS1/Jak2 binding protein or SOCS3/CIS3 almost completely abolished LL-37-induced keratinocyte migration. In conclusion, LL-37 induces keratinocyte migration via heparin-binding-EGF-mediated transactivation of EGFR, and SOCS1/Jak 2 binding and SOCS3/CIS3 negatively regulate this migration. The results of this study suggest that LL-37 closes skin wounds by the induction of keratinocyte migration.