"Rogue" neutrophil-subset [DEspR+CD11b+/CD66b+] immunotype is an actionable therapeutic target for neutrophilic inflammation-mediated tissue injury - studies in human, macaque and rat LPS-inflammation models.
"Rogue" neutrophil-subset [DEspR+CD11b+/CD66b+] immunotype is an actionable therapeutic target for neutrophilic inflammation-mediated tissue injury - studies in human, macaque and rat LPS-inflammation models.
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DOI：
10.3389/fimmu.2022.1008390
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发表时间：
2022
影响因子：
7.3
通讯作者：
中科院分区：
文献类型：
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在急性呼吸窘迫综合征(ARDS)患者中性粒细胞表达双内皮素1/信号肽受体(Despr+CD11b+/CD66b+)与低氧血症严重程度(SF-Ratio)和多器官衰竭(SOFA-Score)的相关性(Rs>；0.7)，提示Despr+中性粒细胞亚群是ARDS的一个可操作的治疗靶点。为了验证这一假设，我们进行了体内研究，以验证Despr+中性粒细胞亚群作为治疗靶点，并测试Despr抑制急性中性粒细胞高炎症模型的有效性。我们在三个物种--人、恒河猴、大鼠--脂多糖(LPS)诱导的急性中性粒细胞炎症中进行了测试，其严重程度随剂量的增加而增加。在短暂性急性肺损伤(ALI)猕猴模型中，采用芯片细胞术检测肺泡灌洗液(BALF)中Despr+CD66b+中性粒细胞的含量，以及短暂性急性肺损伤(ALI)猕猴全血和BALF中Despr+CD11b+中性粒细胞含量。我们在内毒素-ALI猕猴模型和高死亡率内毒素诱导的高血压大鼠脑病模型中检测了抗Despr抗体的体内有效性。芯片细胞仪检测发现，肺泡灌洗液中中性粒细胞总数和Despr+CD66b+中性粒细胞计数较激发前明显增加(P=0.034)，外周血中中性粒细胞计数和中性粒细胞/淋巴细胞比率较激发前明显增加(P<0.05)。在LPS-ALI猕猴模型中，流式细胞仪检测到BALF中的Despr+和Despr[-]中性粒细胞增多，这与中重度低氧血症有关。在测定单剂抗Despr抗体的药代动力学后，一次性给予脂多糖抗Despr抗体可减少低氧血症(P=0.03)和中性粒细胞流入肺泡灌洗液(P=0.0001)。猕猴中性粒细胞的体外活细胞成像检测到在Despr+与Despr[-]中性粒细胞之间有更大的“与硬表面的内在粘连”(P<；0.001)。抗Despr[hu6g8]抗体可抑制Despr+中性粒细胞固有的高黏附性，但对Despr[-]中性粒细胞无明显影响(P<；0.001)。在内毒素脑病大鼠模型中，抗Despr[10a3]抗体治疗提高了大鼠的中位生存期(P=0.0007)，并显示出脑靶向参与和生物效应。检测到分段内毒素攻击后人BALF中Despr+中性粒细胞亚群的增加支持循环中Despr+中性粒细胞计数与ARDS严重程度(SOFA评分)的相关性。在脂多糖诱导的一过性ALI和高死亡率脑病模型中，靶向抑制Despr+CD11b+中性粒细胞亚群的有效性和安全性确定了中性粒细胞介导的继发性组织损伤的潜在治疗靶点。
The correlation (Rs > 0.7) of neutrophils expressing the dual endothelin1/signal peptide receptor (DEspR+CD11b+/CD66b+) with severity of hypoxemia (SF-ratio) and multi-organ failure (SOFA-score) in patients with acute respiratory distress syndrome (ARDS) suggest the hypothesis that the DEspR+ neutrophil-subset is an actionable therapeutic target in ARDS. To test this hypothesis, we conducted in vivo studies to validate DEspR+ neutrophil-subset as therapeutic target and test efficacy of DEspR-inhibition in acute neutrophilic hyperinflammation models. We performed tests in lipopolysaccharide (LPS)-induced acute neutrophilic inflammation in three species – human, rhesus macaque, rat – with increasing dose-dependent severity. We measured DEspR+CD66b+ neutrophils in bronchoalveolar lavage fluid (BALF) in healthy volunteers (HVs) 24-hours after segmental LPS-challenge by ChipCytometry, and DEspR+CD11b+ neutrophils in whole blood and BALF in an LPS-induced transient acute lung injury (ALI) model in macaques. We determined anti-DEspR antibody efficacy in vivo in LPS-ALI macaque model and in high-mortality LPS-induced encephalopathy in hypertensive rats. ChipCytometry detected increased BALF total neutrophil and DEspR+CD66b+ neutrophil counts after segmental LPS-challenge compared to baseline (P =0.034), as well as increased peripheral neutrophil counts and neutrophil-lymphocyte ratio (NLR) compared to pre-LPS level (P <0.05). In the LPS-ALI macaque model, flow cytometry detected increased DEspR+ and DEspR[-] neutrophils in BALF, which was associated with moderate-severe hypoxemia. After determining pharmacokinetics of single-dose anti-DEspR[hu6g8] antibody, one-time pre-LPS anti-DEspR treatment reduced hypoxemia (P =0.03) and neutrophil influx into BALF (P =0.0001) in LPS-ALI vs vehicle mock-treated LPS-ALI macaques. Ex vivo live cell imaging of macaque neutrophils detected greater “intrinsic adhesion to hard-surface” in DEspR+ vs DEspR[-] neutrophils (P <0.001). Anti-DEspR[hu6g8] antibody abrogated intrinsic high adhesion in DEspR+ neutrophils, but not in DEspR[-] neutrophils (P <0.001). In the LPS-encephalopathy rat model, anti-DEspR[10a3] antibody treatment increased median survival (P =0.0007) and exhibited brain target engagement and bioeffects. Detection of increased DEspR+ neutrophil-subset in human BALF after segmental LPS-challenge supports the correlation of circulating DEspR+ neutrophil counts with severity measure (SOFA-score) in ARDS. Efficacy and safety of targeted inhibition of DEspR+CD11b+ neutrophil-subset in LPS-induced transient-ALI and high-mortality encephalopathy models identify a potential therapeutic target for neutrophil-mediated secondary tissue injury.
影响因子：
3.7
作者：
Gautam S;Kirschnek S;Wiesmeier M;Vier J;Häcker G
通讯作者：
Häcker G
影响因子：
3.4
作者：
通讯作者：
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