HIF-1α is essential for effective PMN bacterial killing, antimicrobial peptide production and apoptosis in Pseudomonas aeruginosa keratitis.
HIF-1α is essential for effective PMN bacterial killing, antimicrobial peptide production and apoptosis in Pseudomonas aeruginosa keratitis.
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DOI：
10.1371/journal.ppat.1003457
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发表时间：
2013
期刊：
影响因子：
6.7
通讯作者：
Hazlett LD
中科院分区：
文献类型：
--
作者：
Berger EA;McClellan SA;Vistisen KS;Hazlett LD
缺氧诱导因子（hypoxia inducible factor， HIF）-1α是一种在缺氧条件下控制能量代谢和血管生成的转录因子，是先天免疫的有效调节因子。本文所述的研究检测了HIF-1α在眼部感染铜绿假单胞菌（P.）后BALB/c（耐药，角膜愈合）小鼠疾病消退中的作用。此外，目前的研究主要集中在中性粒细胞（PMN），这是角膜炎的主要细胞浸润。使用siRNA和拮抗剂（17-DMAG），在铜绿假单胞菌感染的BALB/c小鼠中评估HIF-1α的作用。临床评分和裂隙灯摄影显示HIF-1α抑制加重了疾病和角膜破坏。实时RT-PCR、免疫组织化学、ELISA、Greiss和MPO检测、细菌负荷、细胞内杀伤、吞噬和凋亡检测进一步检测了HIF-1α的调控作用。尽管在抑制HIF-1α表达后，促炎细胞因子表达增加，MPO水平增加，但体内研究显示NO产生减少，细菌负荷增加。使用PMN进行的体外研究证明，尽管抑制HIF-1α不影响吞噬，但细菌杀伤和细胞凋亡都受到显著影响，抗菌肽的产生也受到影响。总的来说，数据提供的证据表明，抑制HIF-1α在诱导细菌性角膜炎后，将通常耐药的疾病反应转化为易感（角膜变薄和穿孔）。虽然这种抑制作用似乎不影响PMN的迁移或吞噬，但体内和体外研究表明，转录因子对有效的细菌杀灭、细胞凋亡和抗菌肽的产生至关重要。眼睛感染，尤其是角膜感染，常常导致视力丧失，可能需要角膜移植。铜绿假单胞菌是一种常见的机会性细菌，可感染角膜，特别是在长时间佩戴隐形眼镜的用户中。仅在美国就有3000多万这样的人，他们占角膜细菌感染（角膜炎）病例的40%以上。角膜本身是一种特殊的组织，必须有效地抵抗感染，同时保持完整的视力。我们使用各种技术在动物模型中模拟人眼中可能发生的情况，测试免疫反应的细胞和分子如何对细菌作出反应，导致其被消灭或失明。通过更好地了解免疫系统在感染期间的功能，合理设计更有效的治疗方法，保护患者的视力，是可行的。此外，我们对眼睛免疫反应的不同细胞和分子的了解也可能适用于其他传染病。
Hypoxia-inducible factor (HIF)-1α, is a transcription factor that controls energy metabolism and angiogenesis under hypoxic conditions, and a potent regulator of innate immunity. The studies described herein examined the role of HIF-1α in disease resolution in BALB/c (resistant, cornea heals) mice after ocular infection with Pseudomonas (P.) aeruginosa. Furthermore, the current studies focused on the neutrophil (PMN), the predominant cell infiltrate in keratitis. Using both siRNA and an antagonist (17-DMAG), the role of HIF-1α was assessed in P. aeruginosa-infected BALB/c mice. Clinical score and slit lamp photography indicated HIF-1α inhibition exacerbated disease and corneal destruction. Real time RT-PCR, immunohistochemistry, ELISA, Greiss and MPO assays, bacterial load, intracellular killing, phagocytosis and apoptosis assays further tested the regulatory role of HIF-1α. Despite increased pro-inflammatory cytokine expression and increased MPO levels after knocking down HIF-1α expression, in vivo studies revealed a decrease in NO production and higher bacterial load. In vitro studies using PMN provided evidence that although inhibition of HIF-1α did not affect phagocytosis, both bacterial killing and apoptosis were significantly affected, as was production of antimicrobial peptides. Overall, data provide evidence that inhibition of HIF-1α converts a normally resistant disease response to susceptible (corneal thinning and perforation) after induction of bacterial keratitis. Although this inhibition does not appear to affect PMN transmigration or phagocytosis, both in vivo and in vitro approaches indicate that the transcriptional factor is essential for effective bacterial killing, apoptosis and antimicrobial peptide production. Infections of the eye, especially the cornea, often result in vision loss and may require corneal transplantation. Pseudomonas aeruginosa is a common, opportunistic bacterium that can infect the cornea, especially in extended wear contact lens users. There are more than 30 million such individuals in the US alone and they account for over 40% of corneal bacterial infection (keratitis) cases. The cornea itself is a specialized tissue that must effectively combat infections, while preserving intact visual acuity. Using various techniques to mimic in an animal model, what can occur in the human eye, we test how cells and molecules of the immune response react to the bacteria, leading to either its eradication or blindness. By better understanding the functions of the immune system during infection, rational design of more effective treatments for this disease, which preserve a patient's eyesight, are feasible. In addition, what we have learned about the different cells and molecules of the immune response in the eye may be applicable to other infectious diseases, as well.
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