γ-干扰素预刺激的人脐带间充质干细胞来源的细胞外囊泡减轻大鼠大肠埃希菌诱导的急性肺损伤。

Extracellular Vesicles from Interferon-γ-primed Human Umbilical Cord Mesenchymal Stromal Cells Reduce Escherichia coli-induced Acute Lung Injury in Rats.

机构信息

From the Keenan Research Centre for Biomedical Science (A.K.V., M.J., L.O., S. Gagnon, S. Goyal, R.R., C.M., C.S., C.C.d.S., G.F.C., J.G.L.) Interdepartmental Division of Critical Care Medicine and Department of Critical Care (C.C.d.S., G.F.C., J.G.L.) Department of Anesthesia (G.F.C., J.G.L.), St. Michael's Hospital, University of Toronto, Toronto, Ontario, Canada Department of Chemical Engineering and Applied Chemistry (F.X.G.), University of Toronto, Toronto, Ontario, Canada Department of Chemical Engineering, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, Ontario, Canada (P.Z.C., F.X.G.) Department of Anesthesia, Royal College of Surgeons in Ireland, Dublin, Ireland (G.F.C.) Regenerative Medicine Institute (REMEDI) at CÚRAM Centre for Research in Medical Devices, National University of Ireland Galway, Galway, Ireland (J.G.L.).

出版信息

Anesthesiology. 2019 May;130(5):778-790. doi: 10.1097/ALN.0000000000002655.

DOI:10.1097/ALN.0000000000002655

PMID:30870158

Abstract

BACKGROUND

Human umbilical cord mesenchymal stromal cells possess considerable therapeutic promise for acute respiratory distress syndrome. Umbilical cord mesenchymal stromal cells may exert therapeutic effects via extracellular vesicles, while priming umbilical cord mesenchymal stromal cells may further enhance their effect. The authors investigated whether interferon-γ-primed umbilical cord mesenchymal stromal cells would generate mesenchymal stromal cell-derived extracellular vesicles with enhanced effects in Escherichia coli (E. coli) pneumonia.

METHODS

In a university laboratory, anesthetized adult male Sprague-Dawley rats (n = 8 to 18 per group) underwent intrapulmonary E. coli instillation (5 × 10 colony forming units per kilogram), and were randomized to receive (a) primed mesenchymal stromal cell-derived extracellular vesicles, (b) naïve mesenchymal stromal cell-derived extracellular vesicles (both 100 million mesenchymal stromal cell-derived extracellular vesicles per kilogram), or (c) vehicle. Injury severity and bacterial load were assessed at 48 h. In vitro studies assessed the potential for primed and naïve mesenchymal stromal cell-derived extracellular vesicles to enhance macrophage bacterial phagocytosis and killing.

RESULTS

Survival increased with primed (10 of 11 [91%]) and naïve (8 of 8 [100%]) mesenchymal stromal cell-derived extracellular vesicles compared with vehicle (12 of 18 [66.7%], P = 0.038). Primed-but not naïve-mesenchymal stromal cell-derived extracellular vesicles reduced alveolar-arterial oxygen gradient (422 ± 104, 536 ± 58, 523 ± 68 mm Hg, respectively; P = 0.008), reduced alveolar protein leak (0.7 ± 0.3, 1.4 ± 0.4, 1.5 ± 0.7 mg/ml, respectively; P = 0.003), increased lung mononuclear phagocytes (23.2 ± 6.3, 21.7 ± 5, 16.7 ± 5 respectively; P = 0.025), and reduced alveolar tumor necrosis factor alpha concentrations (29 ± 14.5, 35 ± 12.3, 47.2 ± 6.3 pg/ml, respectively; P = 0.026) compared with vehicle. Primed-but not naïve-mesenchymal stromal cell-derived extracellular vesicles enhanced endothelial nitric oxide synthase production in the injured lung (endothelial nitric oxide synthase/β-actin = 0.77 ± 0.34, 0.25 ± 0.29, 0.21 ± 0.33, respectively; P = 0.005). Both primed and naïve mesenchymal stromal cell-derived extracellular vesicles enhanced E. coli phagocytosis and bacterial killing in human acute monocytic leukemia cell line (THP-1) in vitro (36.9 ± 4, 13.3 ± 8, 0.1 ± 0.01%, respectively; P = 0.0004) compared with vehicle.

CONCLUSIONS

Extracellular vesicles from interferon-γ-primed human umbilical cord mesenchymal stromal cells more effectively attenuated E. coli-induced lung injury compared with extracellular vesicles from naïve mesenchymal stromal cells, potentially via enhanced macrophage phagocytosis and killing of E. coli.

摘要

背景

人脐带间充质基质细胞在急性呼吸窘迫综合征的治疗中有很大的应用前景。间充质基质细胞可能通过细胞外囊泡发挥治疗作用，而间充质基质细胞的预激活可能进一步增强其效果。作者研究了是否干扰素-γ预激活的脐带间充质基质细胞会产生具有增强效果的间充质基质细胞衍生的细胞外囊泡，用于治疗大肠杆菌（E. coli）肺炎。

方法

在一所大学的实验室中，麻醉的成年雄性 Sprague-Dawley 大鼠（每组 8 至 18 只）接受肺内大肠杆菌滴注（每公斤 5×10 个菌落形成单位），并随机接受（a）预激活的间充质基质细胞衍生的细胞外囊泡，（b）未激活的间充质基质细胞衍生的细胞外囊泡（每公斤 1 亿个间充质基质细胞衍生的细胞外囊泡），或（c）载体。在 48 小时评估损伤严重程度和细菌负荷。体外研究评估了预激活和未激活的间充质基质细胞衍生的细胞外囊泡增强巨噬细胞细菌吞噬和杀伤的潜力。

结果

与载体组（18 只大鼠中的 12 只，66.7%）相比，接受预激活（11 只中的 10 只，91%）和未激活（8 只中的 8 只，100%）间充质基质细胞衍生的细胞外囊泡的大鼠存活率更高，差异有统计学意义（P=0.038）。与载体组相比，预激活的间充质基质细胞衍生的细胞外囊泡而非未激活的间充质基质细胞衍生的细胞外囊泡降低了肺泡-动脉氧梯度（分别为 422±104、536±58 和 523±68mmHg，P=0.008），减少了肺泡蛋白渗漏（分别为 0.7±0.3、1.4±0.4 和 1.5±0.7mg/ml，P=0.003），增加了肺单核吞噬细胞（分别为 23.2±6.3、21.7±5 和 16.7±5，P=0.025），并降低了肺泡肿瘤坏死因子-α浓度（分别为 29±14.5、35±12.3 和 47.2±6.3pg/ml，P=0.026）。与载体组相比，预激活的间充质基质细胞衍生的细胞外囊泡而非未激活的间充质基质细胞衍生的细胞外囊泡增强了损伤肺中的内皮型一氧化氮合酶的产生（内皮型一氧化氮合酶/β-肌动蛋白=0.77±0.34、0.25±0.29 和 0.21±0.33，P=0.005）。预激活和未激活的间充质基质细胞衍生的细胞外囊泡均增强了体外人急性单核细胞白血病细胞系（THP-1）中大肠杆菌的吞噬和杀菌作用（分别为 36.9±4%、13.3±8%和 0.1±0.01%，P=0.0004），与载体组相比。