Recombinant Myeloperoxidase as a New Class of Antimicrobial Agents.
Recombinant Myeloperoxidase as a New Class of Antimicrobial Agents.
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DOI：
10.1128/spectrum.00522-21
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发表时间：
2022-02-23
影响因子：
3.7
通讯作者：
Cheng G
中科院分区：
文献类型：
--
作者：
Cao Z;Cheng G
含血红素的过氧化物酶广泛分布于动植物界，通过产生强氧化剂在宿主防御中发挥重要作用。髓过氧化物酶（MPO）是含血红素的过氧化物酶的原型，存在于中性粒细胞和单核细胞中。MPO具有广谱的微生物杀灭作用。MPO的大规模生产困难阻碍了其研究和利用。本研究旨在过量表达重组人髓过氧化物酶，并在体外和体内的杀菌活性进行表征。作为本研究的组成部分，建立了稳定表达重组MPO（rMPO）的人HEK 293细胞系。为了研究rMPO的生物化学和酶学性质以及其杀菌活性，过量表达并纯化了rMPO。在这项研究中，rMPO作为单体分泌到培养基中。rMPO显示与天然MPO相似的酶活性。与天然MPO一样，rMPO能够在低nM水平下杀死广谱微生物，包括革兰氏阴性和阳性细菌和真菌。有趣的是，rMPO可以杀死耐药细菌，使其非常适用于医院感染和混合感染的治疗。rMPO的施用显著降低了由铜绿假单胞菌或耐甲氧西林金黄色葡萄球菌诱导的小鼠肺部感染的发病率和死亡率。在动物安全性试验中，通过尾静脉给予100 nM rMPO未导致任何毒性作用迹象。综上所述，数据表明，从稳定表达的人细胞系中纯化的rMPO是一类新的抗菌剂，具有杀死广谱病原体的能力，包括具有或不具有耐药性的细菌和真菌。在过去的20年里，出现了20多种新的传染病。不幸的是，新的抗微生物治疗剂的发现率要低得多。由耐药微生物引起的感染通常对常规治疗无效，导致长期患病，死亡风险更大，医疗保健费用高昂。目前，这一点在缺乏治愈2019冠状病毒病（COVID-19）的方法方面表现得最好。为了对抗这些不可治疗的微生物，迫切需要发现新类别的抗微生物剂。髓过氧化物酶（MPO）在宿主防御中起重要作用。MPO的大规模生产困难阻碍了其研究和利用。我们已经大规模生产了重组MPO，并对其抗菌活性进行了表征。最重要的是，重组MPO显着降低了铜绿假单胞菌或耐甲氧西林金黄色葡萄球菌诱导的小鼠肺炎的发病率和死亡率。我们的数据表明，重组MPO从人类细胞是一个新的类别的抗菌剂具有广谱的活性。
Heme-containing peroxidases are widely distributed in the animal and plant kingdoms and play an important role in host defense by generating potent oxidants. Myeloperoxidase (MPO), the prototype of heme-containing peroxidases, exists in neutrophils and monocytes. MPO has a broad spectrum of microbial killing. The difficulty of producing MPO at a large scale hinders its study and utilization. This study aimed to overexpress recombinant human MPO and characterize its microbicidal activities in vitro and in vivo. A human HEK293 cell line stably expressing recombinant MPO (rMPO) was established as a component of this study. rMPO was overexpressed and purified for studies on its biochemical and enzymatic properties, as well as its microbicidal activities. In this study, rMPO was secreted into culture medium as a monomer. rMPO revealed enzymatic activity similar to that of native MPO. rMPO, like native MPO, was capable of killing a broad spectrum of microorganisms, including Gram-negative and -positive bacteria and fungi, at low nM levels. Interestingly, rMPO could kill antibiotic-resistant bacteria, making it very useful for treatment of nosocomial infections and mixed infections. The administration of rMPO significantly reduced the morbidity and mortality of murine lung infections induced by Pseudomonas aeruginosa or methicillin-resistant Staphylococcus aureus. In animal safety tests, the administration of 100 nM rMPO via tail vein did not result in any sign of toxic effects. Taken together, the data suggest that rMPO purified from a stably expressing human cell line is a new class of antimicrobial agents with the ability to kill a broad spectrum of pathogens, including bacteria and fungi with or without drug resistance. IMPORTANCE Over the past 2 decades, more than 20 new infectious diseases have emerged. Unfortunately, novel antimicrobial therapeutics are discovered at much lower rates. Infections caused by resistant microorganisms often fail to respond to conventional treatment, resulting in prolonged illness, greater risk of death, and high health care costs. Currently, this is best seen with the lack of a cure for coronavirus disease 2019 (COVID-19). To combat such untreatable microorganisms, there is an urgent need to discover new classes of antimicrobial agents. Myeloperoxidase (MPO) plays an important role in host defense. The difficulty of producing MPO on a large scale hinders its study and utilization. We have produced recombinant MPO at a large scale and have characterized its antimicrobial activities. Most importantly, recombinant MPO significantly reduced the morbidity and mortality of murine pneumonia induced by Pseudomonas aeruginosa or methicillin-resistant Staphylococcus aureus. Our data suggest that recombinant MPO from human cells is a new class of antimicrobials with a broad spectrum of activity.
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