Transcriptional Profiling of Phagocytic Leukocytes and Microglia Reveals a Critical Role for Reactive Oxygen Species in Biofilm Containment during Staphylococcus aureus Craniotomy Infection.
Transcriptional Profiling of Phagocytic Leukocytes and Microglia Reveals a Critical Role for Reactive Oxygen Species in Biofilm Containment during Staphylococcus aureus Craniotomy Infection.
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DOI：
10.4049/jimmunol.2200503
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发表时间：
2022-11-15
期刊：
影响因子：
--
通讯作者：
Kielian T
中科院分区：
文献类型：
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作者：
Menousek J;Horn CM;Heim CE;Van Roy Z;Korshoj LE;Kielian T
开颅术用于治疗各种颅内病变。手术部位感染仍然是开颅手术的并发症，尽管使用预防性抗生素和普遍的无菌预防措施。感染发生在1-3%的手术中，大约一半是由金黄色葡萄球菌（S。金黄色葡萄球菌），其在骨瓣上形成生物膜并且对全身抗生素治疗无效。我们使用了S。aureus-dsRed构建体来比较白细胞和小胶质细胞在体外和体内使用开颅感染的小鼠模型的吞噬能力。此外，应用单细胞RNA测序（scRNA-seq）来确定是否可以在体内鉴定吞噬细胞与非吞噬细胞的转录特征。S.金黄色葡萄球菌在体外的小胶质细胞、巨噬细胞、中性粒细胞和粒细胞髓源性抑制细胞中被吞噬的程度相当;金黄色葡萄球菌是有限的在体内，而其他白细胞群体表现出吞噬活性。scRNA-seq比较吞噬细胞（S. aureus-dsRed+）与非吞噬细胞（S. aureus-dsRed-）白细胞鉴定出在吞噬细胞中富集的经典途径（即，活性氧/氮物质（ROS/RNS）、溶酶体、铁摄取和转运），而非吞噬细胞群体具有增加的核糖体、干扰素和缺氧特征。scRNA-seq还揭示了稳健的ROS谱，这导致在NADPH氧化酶（NOX 2）敲除（KO）小鼠中探索开颅术感染。S.与野生型动物相比，NOX 2 KO中的金黄色葡萄球菌负荷、白细胞募集和细胞内细菌负荷显著增加。总的来说，这些结果强调了吞噬细胞中ROS产生对S。金黄色葡萄球菌生物膜遏制，但不清除，在开颅手术感染。
Craniotomies are performed to treat a variety of intracranial pathology. Surgical site infection remains a complication of craniotomy despite the use of prophylactic antibiotics and universal sterile precautions. Infections occur in 1-3% of procedures, with approximately half caused by Staphylococcus aureus (S. aureus) that forms a biofilm on the bone flap and is recalcitrant to systemic antibiotic therapy. We utilized a S. aureus-dsRed construct to compare the phagocytic capacity of leukocytes and microglia in vitro and in vivo using a mouse model of craniotomy infection. In addition, single-cell RNA sequencing (scRNA-seq) was applied to determine whether a transcriptional signature could be identified for phagocytic vs. non-phagocytic cells in vivo. S. aureus was phagocytosed to equivalent extents in microglia, macrophages, neutrophils, and granulocytic myeloid-derived suppressor cells in vitro; however, microglial uptake of S. aureus was limited in vivo, whereas the other leukocyte populations exhibited phagocytic activity. scRNA-seq comparing the transcriptional signatures of phagocytic (S. aureus-dsRed+) vs. non-phagocytic (S. aureus-dsRed−) leukocytes identified classical pathways enriched in phagocytic cells (i.e., reactive oxygen/nitrogen species (ROS/RNS), lysosome, iron uptake and transport) whereas non-phagocytic populations had increased ribosomal, interferon, and hypoxia signatures. scRNA-seq also revealed a robust ROS profile, which led to the exploration of craniotomy infection in NADPH oxidase (NOX2) knockout (KO) mice. S. aureus burden, leukocyte recruitment, and intracellular bacterial load were significantly increased in NOX2 KO compared to wild-type animals. Collectively, these results highlight the importance of ROS generation in phagocytes for S. aureus biofilm containment, but not clearance, during craniotomy infection.