Activation of human mast cells by retrocyclin and protegrin highlight their immunomodulatory and antimicrobial properties
Activation of human mast cells by retrocyclin and protegrin highlight their immunomodulatory and antimicrobial properties
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DOI：
10.18632/oncotarget.5611
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发表时间：
2015-10-06
期刊：
影响因子：
--
通讯作者：
Ali, Hydar
中科院分区：
文献类型：
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作者：
Gupta, Kshitij;Kotian, Akhil;Ali, Hydar
逆转录周期素(RC-100，RC-101)和前列环素-1(PG-1)抗菌肽(AMPs)的临床前评估非常重要，因为它们具有治疗细菌、真菌和病毒感染的潜力。人肥大细胞(HMCs)在宿主防御和伤口愈合中发挥着重要作用，但其调控功能的研究尚未见报道。在这里，我们报告了化学合成的RC-100和PG-1引起HMCS钙动员和脱颗粒，但这些反应不能被已知的AMPS受体甲酰肽受体样1(FPRL1)的抑制剂所阻断。但RC-100和PG-1可诱导稳定表达Mas相关G蛋白偶联受体X2(MRgX2)的大鼠嗜碱粒白血病(RBL-2H3)细胞脱颗粒。这些AMP的化学合成成本高得令人望而却步，合成后的修饰(环化、二硫键、折叠)不足以达到最佳的抗菌活性。事实上，我们发现通过MRGX2导致肥大细胞脱颗粒的合成RC-100没有显示出任何抗菌活性。绿色荧光蛋白(GFP)标记的RC-101(类似RC-100)和从转基因植物叶绿体中纯化的GFP标记的PG-1杀死细菌并诱导肥大细胞脱颗粒。此外，GFP-PG1可与表达MRgX2的RBL-2H3细胞特异性结合。这些发现表明，逆转录周期素和前列联素独立于FPRL1激活HMCs，但通过MRGX2激活。利用AMPS的这一新特性来激活肥大细胞的宿主防御/伤口愈合特性，以及它们的抗菌活性，扩大了它们的临床潜力。在工厂中低成本生产AMP应该有助于它们进入临床，克服目前生产系统中的主要障碍。
Preclinical evaluation of Retrocyclins (RC-100, RC-101) and Protegrin-1 (PG-1) antimicrobial peptides (AMPs) is important because of their therapeutic potential against bacterial, fungal and viral infections. Human mast cells (HMCs) play important roles in host defense and wound healing but the abilities of retrocyclins and protegrin-1 to harness these functions have not been investigated. Here, we report that chemically synthesized RC-100 and PG-1 caused calcium mobilization and degranulation in HMCs but these responses were not blocked by an inhibitor of formyl peptide receptor-like 1 (FPRL1), a known receptor for AMPs. However, RC-100 and PG-1 induced degranulation in rat basophilic leukemia (RBL-2H3) cells stably expressing Mas related G protein coupled receptor X2 (MrgX2). Chemical synthesis of these AMPs is prohibitively expensive and post-synthesis modifications (cyclization, disulfide bonds, folding) are inadequate for optimal antimicrobial activity. Indeed, we found that synthetic RC-100, which caused mast cell degranulation via MrgX2, did not display any antimicrobial activity. Green-fluorescent protein (GFP)-tagged RC-101 (analog of RC-100) and GFP-tagged PG-1 purified from transgenic plant chloroplasts killed bacteria and induced mast cell degranulation. Furthermore, GFP-PG1 bound specifically to RBL-2H3 cells expressing MrgX2. These findings suggest that retrocyclins and protegrins activate HMCs independently of FPRL1 but via MrgX2. Harnessing this novel feature of AMPs to activate mast cell's host defense/wound healing properties in addition to their antimicrobial activities expands their clinical potential. Low cost production of AMPs in plants should facilitate their advancement to the clinic overcoming major hurdles in current production systems.