TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES最新文献_第2页

Dietary Iron Deficiency Exacerbates Bleomycin-Induced Acute Respiratory Distress Syndrome 膳食铁缺乏加剧博莱霉素诱导的急性呼吸窘迫综合征

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1296

Y. Scindia, E. Rodriguez, M. Wheeler, B. Mehrad

引用次数: 0

Extracellular Heme Impairs Inflammatory Responses of Mononuclear Phagocytes to TLR7 and TLR 8 Stimulation 细胞外血红素损害单核吞噬细胞对TLR7和tlr8刺激的炎症反应

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1313

G. Qu, Y. Scindia, N. Yang, B. Mehrad

{"title":"Extracellular Heme Impairs Inflammatory Responses of Mononuclear Phagocytes to TLR7 and TLR 8 Stimulation","authors":"G. Qu, Y. Scindia, N. Yang, B. Mehrad","doi":"10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1313","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1313","url":null,"abstract":"IntroductionThe COVID-19 pandemic has had an unprecedented impact with over 73 million confirmed cases and over1.6 million deaths globally at the time of this writing. Patients with severe SARS-CoV-2 infection can develop acute respiratory distress syndrome (ARDS). The illness is associated with tissue hemorrhage, causing local hemolysis and the release of heme in the lung. Extracellular heme is rapidly oxidized from Fe2+ to Fe3+, and has been reported to activate MYD88-mediated inflammatory signaling through Toll-like receptor (TLR)-4. SARS-CoV2 is a single strand RNA virus, which is sensed in part by endosomal TLR7/8 for initiating host innate responses. We hypothesized that heme impairs TLR7/8 mediated type 1 interferon response to coronaviruses, thereby worsening the infection. MethodsPeripheral blood mononuclear cells (PBMC) were isolated from each of 6 donor leukocytes, followed by CD14+ isolation. CD14+ monocytes were either studied directly or further differentiated into macrophages under the stimulation of M-CSF. Resiquimod (R848), an agonist for TLR7/8 in humans and TLR7 in a murine systems, was used as a mimic of coronaviruses. CD14+ monocytes and monocytes derived macrophages (MDM) were treated with hemin (heme containing Fe3+ iron), R848, or a combination of hemin and R848. Untreated cells were used as control. The production of proinflammatory cytokines including TNF, IL6, IFN-alpha, and IFN-beta, were measured by ELISA. The expressions of type 1 interferon stimulated genes (x MX1, ISG15, ISG20, and OASL) were assessed via quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Results Hemin alone has no effect on the expression of IL6, TNF, or type 1 interferon-stimulated genes by CD14+ monocytes and MDM, in comparison to untreated cells. In monocytes, hemin resulted in marked inhibition of type-I interferon responses induced by TLR7/8 agonism, but had no effect on IL6 or TNF production. In macrophages, hemin significantly downregulated the production of IL6, but did not affect TNF production or interferon signaling induced by TLR7/8 agonism. Conclusion Heme appears to impair the type 1 interferon response to coronavirus and proinflammatory cytokines productions in human mononuclear phagocytes. Furthermore, heme has different effects on human primary monocytes as compared to macrophages in vitro. We speculate that heme may thus worsens the infection caused by SARS-CoV2 in vivo. Supported by: NIH R01AI135128 and U01EB024501.","PeriodicalId":162721,"journal":{"name":"TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES","volume":"7 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129994792","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Time Course Phagocyte Accumulation in the Lung in Murine Mild Sepsis 轻度脓毒症小鼠肺部吞噬细胞积累的时间过程

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1307

Q. Meng, F. Seto, T. Kariya, T. Hiruma, T. Totsu, K. Uchida

引用次数: 0

Regulatory T Cell Subsets in Pulmonary Fibrosis 肺纤维化中的调节性T细胞亚群

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1305

T. Velez, Y. M. Yoon, K. Blaine, C. Lee, M. Strek, R. Guzy, A. Sperling

引用次数: 0

Effect of Intravenous Immunoglobulin on Neutrophil Antimicrobial and Inflammatory Functions - Implications in Neutrophil-Mediated Immunopathology 静脉注射免疫球蛋白对中性粒细胞抗菌和炎症功能的影响——在中性粒细胞介导的免疫病理中的意义

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1309

J.A. Masso Silva, G. Sakoulas, V. Nizet, L. C. Crotty Alexander, A. Meier

{"title":"Effect of Intravenous Immunoglobulin on Neutrophil Antimicrobial and Inflammatory Functions - Implications in Neutrophil-Mediated Immunopathology","authors":"J.A. Masso Silva, G. Sakoulas, V. Nizet, L. C. Crotty Alexander, A. Meier","doi":"10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1309","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1309","url":null,"abstract":"Rationale. Neutrophils are the most abundant circulating leukocytes and the first line of defense against invading pathogens. Known for their function as phagocytic cells, neutrophils also capture and kill pathogens via elaboration of Neutrophil Extracellular Traps (NETosis), and production of reactive oxygen species (ROS), antimicrobial peptides and proteases. Conversely, neutrophils also have been associated with immunopathology. For example, numerous studies have indicated that neutrophils play a role in acute respiratory distress syndrome (ARDS), which is a major cause of death in COVID-19. We found that COVID-19 patients with ARDS have circulating neutrophils with a hyper-activated phenotype: elevated NETosis, ROS production and phagocytosis. While a small prospective clinical trial showed benefit of intravenous immunoglobulin (IVIG) in mitigating respiratory comorbidity in COVID-19, and prior literature showed benefit of IVIG in viral ARDS, the exact mechanisms are not known. Current clinical therapeutic trials have targeted NET clearance following their release, but not aimed at preventing their formation. In concordance with previous findings, we hypothesized that inhibition of systemic activation of neutrophils with IVIG may reduce collateral tissue damage by blocking NETosis. Methods. Neutrophils from healthy controls were treated ex vivo with IVIG and NETosis, ROS production and phagocytosis quantified. Stimulants phorbol myristate acetate (PMA), for the canonical pathway, and nigericin, for the non-canonical pathway, were used. NETosis was quantified with Quant-iT™ PicoGreen™ dsDNA Assay (Invitrogen) and by NET visualization using myeloperoxidase staining;ROS production was assessed by OxyBURST™ Green H2DCFDA (Invitrogen). Phagocytosis of pHrodo™ Red S. aureus Bioparticles™ (Invitrogen) was also quantified.Results. Ex vivo treatment of healthy neutrophils with IVIG was associated with a significant decrease in NETosis at concentrations of 5 and 10 mg/ml of IVIG (p<0.0001;Figure 1A). IVIG suppression of NETosis was associated with a dose effect. ROS production was also diminished in all concentrations of IVIG tested, including as low as 0.2 mg/ml (p<0.0001;Figure 1B). Phagocytosis was not only preserved but was boosted in the setting of IVIG treatment at 10 mg/ml (p<0.05;Figure 1C).Conclusion. These ex vivo data utilizing fresh human neutrophils demonstrate the potential beneficial effect of IVIG, an FDAapproved treatment, in the context of neutrophil-mediated immunopathology. IVIG may be a potential treatment to prevent progression of vascular and lung inflammation and damage in COVID-19, consistent with emerging evidence of its therapeutic benefit in this disorder. Results from an FDA Phase 3 prospective randomized international study that evaluates IVIG in COVID-19 are highly anticipated.","PeriodicalId":162721,"journal":{"name":"TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES","volume":"20 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115422242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tyrosine Kinase Inhibitor Imatinib Alleviates the Progression of Ventilator-Induced Lung Injury 酪氨酸激酶抑制剂伊马替尼缓解呼吸机所致肺损伤的进展

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1301

Y. Xin, M. Cereda, K. Martin, S. Humayun, I. Duncan, H. Profka, S. Kadlecek, R. Rizi

引用次数: 0

RLS-0071 Inhibits Acute Lung Injury in a Novel 2-Hit Model RLS-0071抑制急性肺损伤的新型2-Hit模型

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1312

P. Kumar, N. Krishna, A. Sampson, B. Lassiter, M. Gregory Rivera, P. Hair, K. Jackson, A. Enos, A. Werner, C. Goldberg, F. Lattanzio, K. Cunnion, M. Glesby

{"title":"RLS-0071 Inhibits Acute Lung Injury in a Novel 2-Hit Model","authors":"P. Kumar, N. Krishna, A. Sampson, B. Lassiter, M. Gregory Rivera, P. Hair, K. Jackson, A. Enos, A. Werner, C. Goldberg, F. Lattanzio, K. Cunnion, M. Glesby","doi":"10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1312","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1312","url":null,"abstract":"Rationale: Acute lung injury (ALI) is often a complication of viral infection (SARS-CoV-2, Influenza, RSV) that leads to significant morbidity and mortality. A novel rat two-hit ALI model was developed that uses a complement stimulus for the second hit, leading to a robust immune-mediated ALI, characterized by leukopenia, neutrophil infiltration of the lung tissue as well as neutrophil extracellular trap (NETs) formation and cytokine storm. ReAlta's lead compound, RLS-0071, is a PEGylated peptide with dual acting anti-inflammatory activity that inhibits classical and lectin complement activation, as well as neutrophil-mediated NETosis in vitro. The efficacy of RLS-0071 on ALI, complement activation, NET formation and inflammatory cytokine release was assessed in this preclinical animal model. Methods: Wistar rats were infused intravenously with lipopolysaccharide (LPS) (1st hit), followed by incompatible red blood cells (RBC) 30 min later (2-hit). At 4 hours, blood was collected to assess C5a levels, free DNA (NET biomarker) and cytokine levels. Lung tissue was recovered for histological analysis. RLS-0071 administration prior to RBC transfusion (prophylaxis dosing) and at fixed times (30 seconds to 180 minutes) after 2-hit (rescue dosing) was examined to assess the impact of the compound on lung histology, C5a, free DNA as well as inflammatory cytokine levels. Results: Scoring of hematoxylin and eosin (H&E) lung sections showed a significant reduction in lung injury with single prophylaxis dose of RLS-0071 given 30 minutes before the second hit preserving lung histology in the animals. Analysis of the sections in which the images were converted to black and white pixels quantified by ImageJ (NIH) were used to derive a lung injury score (0-5) as shown below. RLS-0071 delivered as prophylaxis or rescue significantly reduced neutrophil infiltration, C5a levels, free DNA levels in the blood and pro inflammatory cytokines IL-1b, IL-6, TNFα and IL-17. Rescue dosing demonstrated that RLS-0071 could significantly prevent lung damage when delivered up to 180 minutes after the 2-hit. Conclusion: Animals receiving RLS-0071 showed lung tissue architecture comparable to controls. RLS-0071 demonstrated the ability to prevent downstream C5a proinflammatory signaling via complement modulation consistent with the observed reduction of pro-inflammatory cytokines production such as IL-1, IL-6, and TNFα. RLS-0071 further modulates the innate inflammatory response by inhibiting NETosis displaying synergistic mechanism of actions in this rat model of ALI.","PeriodicalId":162721,"journal":{"name":"TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES","volume":"17 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122257903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Extracellular Specks, a Marker of NLRP3 Inflammasome Activation 细胞外斑点:NLRP3炎性小体激活的标志

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1318

M. Rhedin, Z. Jevnikar Rojnik, B. Collins, J. Jirholt

引用次数: 0

Mechanosensation in Macrophages Protects the Lung from Infection-Associated Lung Injury via TRPV4-Rac1-MAPK Axis 巨噬细胞的机械感觉通过TRPV4-Rac1-MAPK轴保护肺部免受感染相关的肺损伤

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1308

R. Scheraga, S. Abraham, L. Grove, B. Southern, J. Hasday, M. Olman

引用次数: 0

Intravascular Infusible Extracellular Matrix for the Mitigation of Severe Systemic Inflammation Relevant to Sepsis and COVID-19 Pathology 血管内不溶性细胞外基质缓解与败血症和COVID-19病理相关的严重全身炎症

TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES Pub Date : 1900-01-01 DOI: 10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1322

R. Wang, A. Lyons, R. Middleton, M. Hepokoski, K. Christman

{"title":"Intravascular Infusible Extracellular Matrix for the Mitigation of Severe Systemic Inflammation Relevant to Sepsis and COVID-19 Pathology","authors":"R. Wang, A. Lyons, R. Middleton, M. Hepokoski, K. Christman","doi":"10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1322","DOIUrl":"https://doi.org/10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a1322","url":null,"abstract":"Rationale: Multi-organ failure (MOF), associated with conditions such as acute respiratory distress syndrome, sepsis, and the recent SARS-CoV-2 virus, often exacerbates disease severity by the overwhelming systemic inflammation, thus, highlighting the need for alternative therapeutics. To address this need, intravascularly delivered infusible extracellular matrix (iECM) material was evaluated in a sepsis mouse model using lipopolysaccharide (LPS). Immunomodulatory properties of decellularized ECM that have demonstrated endogenous tissue repair promotion could favorably mitigate systemic inflammation pathology related to MOF. Compared to decellularized ECM or injectable ECM hydrogels where deployment is limited to a specific tissue per procedure, intravascular iECM delivery targets leaky vasculature in areas of inflammation. We hypothesized iECM could simultaneously alleviate systemic inflammation across multiple vulnerable tissues, thus, demonstrating potential to mitigate MOF in systemic inflammatory conditions. Methods: iECM was generated as previously described1. Porcine left ventricular tissue was decellularized, lyophilized, milled into fine powder, and 2-day pepsin digested. Insoluble ECM proteins were pelleted by high-speed centrifugation, supernatant was dialyzed in 100- 500 Da tubing, and dialyzed ECM was lyophilized, resuspended with 1x PBS, and sterile filtered. Resulting iECM was aliquoted and lyophilized for storage until resuspended with deionized water. A dual LPS-LPS mouse was utilized for modeling severe systemic inflammation. Nine-week female C57BL6/J mice underwent the following procedures: 1) Intraperitoneal (IP) injection of LPS (10 mg/kg), 2) IP LPS injection (1 mg/kg) at 6 hours for further prolonged and severe inflammation, 3) 200 μL tail vein injection of saline or iECM (10 mg/mL) at 10 hours (n = 6 per group), and 4) euthanasia and harvesting of lungs, heart, brain, and kidneys for gene expression analysis by qRT-PCR and Nanostring nCounter® Immunology Panel (Mouse). Results: Il1b and Il6 trended (p < 0.1) lower for the lungs while both were significantly decreased for the heart following iECM delivery. Additionally, Il6 was significantly decreased in the brain and kidney (Fig 1A). Multiplex profiling of lungs demonstrated inflammatory marker downregulation, including Il6, Il1α, and various chemokines (Fig 1B). Gene enrichment determined modulation of inflammatory cytokine profiles along with immune cell activation, proliferation, migration, and differentiation. Similar results were determined for the heart, brain, and kidney (data not shown). Conclusion: Systemic immunomodulation was observed among multiple affected organs, demonstrating iECM's potential for mitigating systemic inflammatory responses. Future work will investigate infiltrating immune cell subpopulations, validate results through proteomic analysis, and complete long term tissue morphology and functional assessments.","PeriodicalId":162721,"journal":{"name":"TP4. TP004 ARDS, SEPSIS, AND TRANSPLANTATION MECHANISTIC STUDIES","volume":"75 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121085789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0