Infiltration of inflammatory macrophages and neutrophils and widespread pyroptosis in lung drive influenza lethality in nonhuman primates.
Infiltration of inflammatory macrophages and neutrophils and widespread pyroptosis in lung drive influenza lethality in nonhuman primates.
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DOI：
10.1371/journal.ppat.1010395
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发表时间：
2022-03
期刊：
影响因子：
6.7
通讯作者：
Barratt-Boyes SM
中科院分区：
文献类型：
--
作者：
Corry J;Kettenburg G;Upadhyay AA;Wallace M;Marti MM;Wonderlich ER;Bissel SJ;Goss K;Sturgeon TJ;Watkins SC;Reed DS;Bosinger SE;Barratt-Boyes SM
严重的流感每年导致数万人死亡，但对人类致命性的机制知之甚少。在这里，我们使用了一个独特的翻译模型，致命的H5 N1流感在食蟹猴，利用吸入小颗粒病毒气溶胶，以确定驱动致命疾病的机制。肺组织的RNA测序显示，在感染后两天内出现强烈的干扰素反应，导致干扰素刺激基因的广泛表达，包括炎性细胞因子和趋化因子。患有致死性疾病的猕猴具有肺泡巨噬细胞（AM）的快速和深刻的损失以及活化的CCR 2 + CX 3CR 1+间质巨噬细胞（IM）和中性粒细胞向肺中的浸润。与轻度流感猕猴相比，支气管肺泡灌洗（BAL）中AM和中性粒细胞的平行变化与病毒载量相关。致死性流感中的AM和IM均为M1型炎性巨噬细胞，其表达中性粒细胞趋化因子，而中性粒细胞表达与中性粒细胞胞外陷阱（NETs）的激活和产生相关的基因。NETs在肺中突出，并且在肺泡腔以及肺实质中被发现。肺组织中与细胞凋亡相关的基因表达增加，支气管肺泡灌洗液和肺组织匀浆中存在活化的炎性半胱氨酸蛋白酶、IL-1β和裂解的gasdermin D（GSDMD）。切割的GSDMD由肺巨噬细胞和肺泡上皮细胞表达，这些细胞大量存在于肺泡腔中，与上皮完整性丧失一致。裂解的GSDMD与肺泡中的病毒NP表达细胞共定位，反映了感染细胞的焦亡。这些新的发现揭示了肺中与炎性巨噬细胞和中性粒细胞浸润、NET的加工和细胞死亡相关的强效干扰素和炎性级联反应介导了非人灵长类动物和人类中的致命H5 N1流感。这些先天途径代表了预防人类严重流感和潜在的其他原发性病毒性肺炎的有希望的治疗靶点。流感可导致急性肺损伤和死亡，但导致人类致命流感的机制尚不清楚。我们使用了一种新型的非人灵长类动物致命流感模型，该模型由高致病性禽流感病毒气溶胶感染引起，与人类疾病非常相似，以定义该病毒如何导致严重肺炎。我们发现，一种强有力的先天免疫反应始于肺部高水平的干扰素和炎症因子的产生，从而导致严重的疾病。由于这种早期反应，包括巨噬细胞和中性粒细胞在内的炎性细胞被募集到肺中，这反过来导致释放阻塞肺泡的中性粒细胞胞外陷阱。此外，在致命性流感期间，肺中发生了一种称为焦亡的细胞死亡的特别炎症形式。这些新的发现表明，强烈的干扰素反应导致巨噬细胞和中性粒细胞的炎症级联反应，释放中性粒细胞胞外陷阱，并通过焦亡导致细胞死亡，是致命流感中急性肺损伤的原因。这些先天途径可以被药物靶向，以防止重症流感患者的肺损伤。
Severe influenza kills tens of thousands of individuals each year, yet the mechanisms driving lethality in humans are poorly understood. Here we used a unique translational model of lethal H5N1 influenza in cynomolgus macaques that utilizes inhalation of small-particle virus aerosols to define mechanisms driving lethal disease. RNA sequencing of lung tissue revealed an intense interferon response within two days of infection that resulted in widespread expression of interferon-stimulated genes, including inflammatory cytokines and chemokines. Macaques with lethal disease had rapid and profound loss of alveolar macrophages (AMs) and infiltration of activated CCR2+ CX3CR1+ interstitial macrophages (IMs) and neutrophils into lungs. Parallel changes of AMs and neutrophils in bronchoalveolar lavage (BAL) correlated with virus load when compared to macaques with mild influenza. Both AMs and IMs in lethal influenza were M1-type inflammatory macrophages which expressed neutrophil chemotactic factors, while neutrophils expressed genes associated with activation and generation of neutrophil extracellular traps (NETs). NETs were prominent in lung and were found in alveolar spaces as well as lung parenchyma. Genes associated with pyroptosis but not apoptosis were increased in lung, and activated inflammatory caspases, IL-1β and cleaved gasdermin D (GSDMD) were present in bronchoalveolar lavage fluid and lung homogenates. Cleaved GSDMD was expressed by lung macrophages and alveolar epithelial cells which were present in large numbers in alveolar spaces, consistent with loss of epithelial integrity. Cleaved GSDMD colocalized with viral NP-expressing cells in alveoli, reflecting pyroptosis of infected cells. These novel findings reveal that a potent interferon and inflammatory cascade in lung associated with infiltration of inflammatory macrophages and neutrophils, elaboration of NETs and cell death by pyroptosis mediates lethal H5N1 influenza in nonhuman primates, and by extension humans. These innate pathways represent promising therapeutic targets to prevent severe influenza and potentially other primary viral pneumonias in humans. Influenza can cause acute lung injury and death, but the mechanisms resulting in lethal influenza in humans are not well understood. We used a novel model of lethal influenza in nonhuman primates caused by aerosol infection with highly pathogenic avian influenza virus that closely resembles human disease to define how the virus causes severe pneumonia. We found that a potent innate immune response starting with high-level production of interferons and inflammatory factors in the lung drives severe disease. Inflammatory cells including macrophages and neutrophils were recruited into lung because of this early response, which in turn led to release of neutrophil extracellular traps that blocked lung alveoli. In addition, a particularly inflammatory form of cell death known as pyroptosis occurred in lungs during lethal influenza. These new findings show that an intense interferon response leading to an inflammatory cascade of macrophages and neutrophils, release of neutrophil extracellular traps, and cell death by pyroptosis is responsible for acute lung injury in lethal influenza. These innate pathways could be targeted by drugs to prevent lung injury in critically ill influenza patients.
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影响因子：
13.6
作者：
Ghari F;Quirke AM;Munro S;Kawalkowska J;Picaud S;McGouran J;Subramanian V;Muth A;Williams R;Kessler B;Thompson PR;Fillipakopoulos P;Knapp S;Venables PJ;La Thangue NB
通讯作者：
La Thangue NB
人类感染新型禽源甲型流感 (H7N9) 病毒。
影响因子：
158.5
作者：
Gao, Rongbao;Cao, Bin;Shu, Yuelong
通讯作者：
Shu, Yuelong
DOI：
10.4049/jimmunol.1302269
发表时间：
2014-03-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Cai Y;Sugimoto C;Arainga M;Alvarez X;Didier ES;Kuroda MJ
通讯作者：
Kuroda MJ
影响因子：
5.2
作者：
Han, Yi;Zhang, Haidong;Gu, Guorong
通讯作者：
Gu, Guorong
影响因子：
7.3
作者：
Cortjens, Bart;de Boer, Onno J.;Bem, Reinout A.
通讯作者：
Bem, Reinout A.