Inflammation Program and Department of Internal Medicine Roy J. and Lucille A Carver College of Medicine, University of Iowa and Veterans Administration Medical Center, Iowa City, Iowa, USA.
Department of Microbiology and Immunology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa, USA.
J Leukoc Biol. 2020 Nov;108(5):1543-1553. doi: 10.1002/JLB.4MA0820-649R. Epub 2020 Sep 25.
Optimal innate immune response to infection includes eradication of potential pathogens, resolution of associated inflammation, and restitution of homeostasis. Phagocytosing human polymorphonuclear leukocytes (hPMN) undergo accelerated apoptosis, a process referred to as phagocytosis-induced cell death (PICD) and an early step in their clearance from inflammatory sites. Among human pathogens that modulate hPMN apoptosis, Neisseria gonorrhoeae delays PICD, which may contribute to the exuberant neutrophilic inflammation that characterizes gonorrhea. To elucidate the mechanisms underlying delayed PICD, we compared features of hPMN cell death that followed phagocytosis of N. gonorrhoeae FA1090 wild-type (GC) or serum-opsonized zymosan (OPZ), a prototypical stimulus of PICD. Phosphatidylserine externalization required NADPH oxidase activity after ingestion of GC or OPZ, and annexin V staining and DNA fragmentation were less after phagocytosis of GC compared to OPZ. Caspase 3/7 and caspase 9 activities after phagocytosis of GC were less than that seen after ingestion of OPZ, but caspase 8 activity was the same after ingestion of GC or OPZ. When hPMN sequentially ingested GC followed by OPZ, both caspase 3/7 and 9 activities were less than that seen after OPZ alone, and the inhibition was dose dependent for GC, suggesting that ingestion of GC actively inhibited PICD. Sequential phagocytosis did not block caspase 8 activity, mitochondrial depolarization, or annexin V/propidium iodide staining compared to responses of hPMN fed OPZ alone, despite inhibition of caspases 3/7 and 9. Taken together, these data suggest that active inhibition of the intrinsic pathway of apoptosis contributes to the delay in PICD after hPMN ingestion of N. gonorrhoeae.
最佳的先天免疫反应包括消除潜在的病原体、解决相关炎症和恢复体内平衡。吞噬人类多形核白细胞(hPMN)会加速细胞凋亡,这个过程被称为吞噬诱导的细胞死亡(PICD),是它们从炎症部位清除的早期步骤。在调节 hPMN 凋亡的人类病原体中,淋病奈瑟菌(Neisseria gonorrhoeae)延迟了 PICD,这可能导致淋病特征性的过度中性粒细胞炎症。为了阐明延迟 PICD 的机制,我们比较了吞噬淋病奈瑟菌 FA1090 野生型(GC)或血清调理的酵母聚糖(OPZ)后 hPMN 死亡的特征,OPZ 是 PICD 的典型刺激物。吞噬 GC 或 OPZ 后需要 NADPH 氧化酶活性来实现磷脂酰丝氨酸外翻,并且吞噬 GC 后 Annexin V 染色和 DNA 片段化比吞噬 OPZ 后少。吞噬 GC 后 caspase 3/7 和 caspase 9 的活性低于吞噬 OPZ 后,但吞噬 GC 或 OPZ 后 caspase 8 的活性相同。当 hPMN 顺序吞噬 GC 后再吞噬 OPZ 时,caspase 3/7 和 caspase 9 的活性均低于单独吞噬 OPZ 时的活性,并且 GC 的抑制作用呈剂量依赖性,表明吞噬 GC 主动抑制了 PICD。与单独用 OPZ 喂养的 hPMN 反应相比,顺序吞噬并没有阻断 caspase 8 活性、线粒体去极化或 Annexin V/碘化丙啶染色,尽管 caspase 3/7 和 9 被抑制。总之,这些数据表明,吞噬淋病奈瑟菌后,内源性凋亡途径的主动抑制有助于延迟 PICD。