scholarly journals Osteopontin Mediates Murine Transfusion-Related Acute Lung Injury through Stimulation of Pulmonary Neutrophil Accumulation

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 739-739
Author(s):  
Rick Kapur ◽  
Gopinath Kasetty ◽  
Johan Rebetz ◽  
Arne Egesten ◽  
John W Semple
Keyword(s):  
Acute Lung Injury ◽  
Blood Transfusion ◽  
Lung Injury ◽  
Mhc Class I ◽  
Class I ◽  
Knock Out ◽  
Wide Range ◽  
Knock Out Mice ◽  
Stimulation Of

Abstract Transfusion-related acute lung injury (TRALI) is a syndrome of respiratory distress which develops within 6 hours of blood transfusion. It is the leading cause of transfusion-related deaths and the pathogenesis is complex and incompletely understood. In the majority of the cases, anti-leukocyte antibodies present in the transfused blood product, in combination with recipient predisposing risk-factors such as inflammation, are implicated to be responsible for the onset of TRALI. Unfortunately, no therapies are available for TRALI. Osteopontin (OPN) is an extracellular matrix protein with multiple biological functions. OPN is involved in normal physiological processes, such as cell migration and adhesion, but has also been implicated in a wide range of disease states, including cancer, atherosclerosis, glomerulonephritis, and several chronic inflammatory diseases. Interestingly, OPN is upregulated at sites of inflammation and tissue remodeling. As inflammation is an important risk factor for TRALI development, and as neutrophils (PMNs) are known effector cells in the pathogenesis of TRALI which migrate and accumulate in the lungs during TRALI development, we investigated the potential contribution of OPN in the onset of antibody-mediated TRALI. We utilized a previously established murine TRALI model (Kapur et al, Blood 2017, Blood Advances 2018) in which C57BL/6 mice were first primed with a low dose of lipopolysaccharide (LPS) and depleted of their CD4+ T cells in vivo followed by injection of anti-major histocompatibility complex (MHC) class I antibodies (clones 34-1-2s and AF6-88.5.5.3). The TRALI response was analyzed after 90 minutes by analysis of pulmonary edema (lung wet-to-dry weight ratios, W/Ds) and the levels of pulmonary neutrophils. Wildtype (WT) mice suffered from antibody-mediated TRALI compared to untreated naïve mice, as was shown by their significantly increased lung W/Ds (4.72 vs 4.50, respectively, P<0.0001). This also corresponded to significantly increased levels of pulmonary PMNs compared to untreated naïve mice (34% vs 5%, P<0.0001). In contrast, C57BL/6 OPN knock-out mice were resistant to antibody-mediated TRALI induction as they did not display any significant increase in lung W/Ds levels or pulmonary PMNs compared to untreated naïve OPN knock-out mice (lung W/Ds 4.83 vs 4.75, and pulmonary PMNs 18% vs 16%, respectively). Strikingly, administration of purified recombinant OPN during TRALI induction in C57BL/6 OPN knock-out mice, significantly induced a TRALI reaction (lung W/Ds 5.12 vs 4.75 as compared to untreated naïve OPN knock-out mice, P<0.05). Mechanistically, this TRALI inducing effect of OPN administration to OPN knock-out mice was associated with increased levels of pulmonary PMNs (38% vs 16%, as compared to untreated naïve OPN knock-out mice, P<0.0001). In vivo blocking of OPN in WT mice with an anti-OPN antibody demonstrated decreased lung W/Ds as compared to treatment with an isotype antibody (4.48 vs 4.69, respectively, P<0.05). The OPN blocking response during TRALI was associated with a decreased level of pulmonary PMN accumulation as compared to treatment with an isotype antibody (14% vs 34%, respectively, P<0.0001). As the PMN-chemoattractant macrophage inflammatory protein (MIP)-2 has previously been described to be upregulated in murine antibody-mediated TRALI, we investigated if the OPN-associated pulmonary PMN accumulation and TRALI induction could be related to the levels of MIP-2. We found that plasma MIP-2 levels were increased in mice that were infused with anti-MHC class I antibodies as compared to naïve controls, but that addition or blocking of OPN did not affect these MIP-2 levels. This indicates that the OPN-related PMN responses in TRALI are independent of plasma MIP-2 levels. Collectively, these data indicate OPN as a novel pathogenic factor which enhances antibody-mediated murine TRALI through stimulation of PMN migration towards the lungs, independent of MIP-2. This may suggest that blocking OPN (using an anti-OPN antibody) may prevent TRALI by impairing pulmonary PMN accumulation and could be a therapeutic avenue to explore in combatting this serious adverse complication of blood transfusion. Disclosures No relevant conflicts of interest to declare.

A Two-Event In Vivo Model of Transfusion-Related Acute Lung Injury.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 19-19 ◽  
Author(s):  
Christopher C. Silliman ◽  
Marguerite Kelher ◽  
Tomohiko Masuno ◽  
Ernest E. Moore ◽  
Sagar Damle ◽  
...  
Keyword(s):  
Acute Lung Injury ◽  
Lung Injury ◽  
Mhc Class I ◽  
Mhc Class Ii ◽  
Normal Saline ◽  
Class Ii ◽  
Class I ◽  
In Vivo Model ◽  
Blue Dye

Abstract TRALI is the most common cause of transfusion-related death in the US, and the pathogenesis is related to the infusion of donor anti-leukocyte antibodies or biologic response modifiers (BRMs) including lipids that accumulate during storage of cellular components. We hypothesize that TRALI is the result of two distinct events: the first related to the clinical condition of the patient resulting in pulmonary endothelial activation and sequestration of PMNs and the second is the infusion of antibodies or BRMs along with the transfused product. Methods: PRBCs were obtained from 5 donors and 50% were pre-storage leukoreduced by filtration and the other 50% left as a control, and both stored per AABB criteria. Plasma samples were obtained serially from these units and was heat-treated (56°C for 30 min) to destroy fibrinogen and complement prior to administration. Antibodies to antigens present on leukocytes from Sprague Dawley rats including MHC class I: OX18 & OX27, MHC class II: OX3 & OX6 and anti-granulocyte (PMN) antibodies were obtained commercially. Male rats were given saline (NS) or 2 mg/kg IP of endotoxin (LPS S.enteritidies, non-lethal), incubated for 2 hrs, anesthetized with pentobarbital, the femoral vessels were cannulated, and 10% of the blood volume was withdrawn over 15 min. Plasma from day 1 (10% final) and day 42 (5–10%) PRBCs and 10% LR-PRBCs, and 50 or 100 μg of antibodies (500μl of sera, anti-PMN) were infused over 30 min, followed by IV Evan’s Blue dye (30 mg/kg; 1ml) that binds to albumin. At 6 hours, plasma and bronchoalveolar lavage (BAL) fluid were obtained to determine the % of Evan’s Blue leak into the BAL at 620 nm. Mortality was < 5%. Acute lung injury (ALI) was precipitated in LPS-treated animals by day 42 PRBC plasma (5% & 10%), 10% day 42 LR-PRBC plasma and antibodies to MHC class I antigens (Table). With NS as the first event, rats did not evidence ALI for all groups, including MHC class I antibodies. Moreover, in LPS pre-treated rats, second events consisting of NS, day 0 PRBC, day 0 LR-PRBC plasma, antibodies to MHC class II antigens (OX3 & OX6) and anti-PMN antibodies did not elicit ALI (Table). We conclude that 1) this in vivo model approximates the mortality of the clinical condition, 2) it demonstrates that the pathogenesis requires two events to elicit antibody-induced or BRM-mediated TRALI, and 3) ALI as the result of LPS/MHC class I antibodies evidences a dose-response. ALI as a a Function of Evans Blue Dye Leak 1st Event ⇒ Normal Saline NS LPS 2nd Event ⇓ †=p<.05 vs. 1st event or 2nd event Normal Saline 0.08±0.03 0.24±0.11 MHC Class I OX18 50μg 0.06±0.06 0.18±0.03 MHC Class I OX18 100μg 0.17 1.91±0.7] MHC Class I OX27 50μg 0.19±0.04 1.26±0.1† MHC Class II OX3 50μg 0.07 0.4 MHC Class II OX6 50μg 0.07±0.07 0.2±0.07 Anti-Granulocyte serum 500μl 0.25±0.14 0.22±0.17 Anti-Granulocyte 100μg 0.174 0.09±.02 PRBCs day 1 [10%] 0.10±0.08 0.25±0.09 PRBCs day 42 [5%] 0.13±0.07 2.48±0.46† PRBCs day 42 [10%] 0.16±0.10 1.16±0.34† LR-PRBCs day 1 [10%] 0.16±0.09 0.19±0.05 LR-PRBCs day 42 [10%] 0.20±0.12 2.69±0.58†

Identification of a Two-Step Mechanism Responsible for Antibody-Mediated Transfusion Related Acute Lung Injury (TRALI)

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 846-846
Author(s):  
Christopher G.J. McKenzie ◽  
Michael Kim ◽  
Tarandeep Singh ◽  
John W. Semple
Keyword(s):  
Reactive Oxygen Species ◽  
Acute Lung Injury ◽  
Lung Injury ◽  
Mhc Class I ◽  
Reactive Oxygen ◽  
Lung Damage ◽  
Class I ◽  
Oxygen Species ◽  
Neutrophil Accumulation

Abstract Abstract 846 Transfusion-related acute lung injury (TRALI) is one of the leading causes of transfusion fatalities, and most TRALI reactions are thought to be caused by donor antibodies. It is currently thought that the donor antibodies activate pulmonary neutrophils to produce reactive oxygen species that damage lung tissue. There have been several animal models of TRALI developed including, for example, ex vivo lung models demonstrating the importance of human anti-neutrophil antibodies in TRALI, and in vivo models showing how biological response modifiers can induce recipient lung damage. An in vivo murine model of antibody-mediated TRALI was developed in 2006, and has also shown several similarities with human TRALI induction (Looney MR et al., J Clin Invest 116: 1615, 2006). Specifically, a monoclonal anti-mouse MHC class I antibody (34-1-2s) causes significant increases in excess lung water, lung vascular permeability and mortality within 2 hours after administration. These adverse reactions were found to be due to the antibody's ability to activate pulmonary neutrophils to produce reactive oxygen species (ROS) in an Fc receptor (FcR)-dependent manner. In contrast, however, it was recently shown that 34-1-2s induces pulmonary damage by activating macrophages to generate ROS in a complement (C5a)-dependent process (Strait RT J et al., Exp Med 208: 2525, 2011). In order to better understand this apparent controversy, we attempted to determine the nature of how 34-1-2s mediates its lung damaging properties. 34-1-2s was digested with pepsin or papain to produce F(ab')2 or Fc fragments respectively, and the fragments were tested for their ability to mediate TRALI reactions. In control mice, when intact 34-1-2s antibody was intravenously injected into either CB.17 mice with severe combined immunodeficiency or C5 deficient DBA/2 mice, increased shock, serum MIP-2 (murine equivalent to human IL-8) levels, pulmonary neutrophil accumulation, pulmonary edema and mortality all occurred within 2 hours. In contrast, however, injection with 34-1-2s F(ab')2 fragments was only able to generate MIP-2 production and pulmonary neutrophil accumulation; no lung damage or mortality occurred. Injection of 34-1-2s Fc fragments either alone or together with equal molar concentrations of F(ab')2 fragments failed to induce any lung damage or mortality. These results suggest that 34-1-2s recognition of it's cognate MHC class I antigen may be a priming reaction that stimulates MIP-2 and chemotaxis of neutrophils to the lungs, whereas the Fc portion of the intact molecule is responsible for the second step of exacerbating TRALI symptoms in a complement independent manner. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Plasma from stored packed red blood cells and MHC class I antibodies causes acute lung injury in a 2-event in vivo rat model

Blood ◽  
2009 ◽  
Vol 113 (9) ◽  
pp. 2079-2087 ◽  
Author(s):  
Marguerite R. Kelher ◽  
Tomhiko Masuno ◽  
Ernest E. Moore ◽  
Sagar Damle ◽  
Xianzhong Meng ◽  
...  
Keyword(s):  
Acute Lung Injury ◽  
Lung Injury ◽  
Red Blood Cells ◽  
Mhc Class I ◽  
Rat Model ◽  
Blood Cells ◽  
Class I ◽  
Packed Red Blood Cells ◽  
Stored Blood

Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion death. We hypothesize that TRALI requires 2 events: (1) the clinical condition of the patient and (2) the infusion of antibodies against MHC class I antigens or the plasma from stored blood. A 2-event rat model was developed with saline (NS) or endotoxin (LPS) as the first event and the infusion of plasma from packed red blood cells (PRBCs) or antibodies (OX18 and OX27) against MHC class I antigens as the second event. ALI was determined by Evans blue dye leak from the plasma to the bronchoalveolar lavage fluid (BALF), protein and CINC-1 concentrations in the BALF, and the lung histology. NS-treated rats did not evidence ALI with any second events, and LPS did not cause ALI. LPS-treated animals demonstrated ALI in response to plasma from stored PRBCs, both prestorage leukoreduced and unmodified, and to OX18 and OX27, all in a concentration-dependent fashion. ALI was neutrophil (PMN) dependent, and OX18/OX27 localized to the PMN surface in vivo and primed the oxidase of rat PMNs. We conclude that TRALI is the result of 2 events with the second events consisting of the plasma from stored blood and antibodies that prime PMNs.

scholarly journals RB depletion is required for the continuous growth of tumors initiated by loss of RB

PLoS Genetics ◽  
2021 ◽  
Vol 17 (12) ◽  
pp. e1009941
Author(s):  
Alex Doan ◽  
Julia Arand ◽  
Diana Gong ◽  
Alexandros P. Drainas ◽  
Yan Ting Shue ◽  
...  
Keyword(s):  
Mouse Model ◽  
Cancer Cells ◽  
Mechanisms Of Action ◽  
Thyroid Tumors ◽  
Continuous Growth ◽  
Knock Out ◽  
Cycle Arrest ◽  
Wide Range ◽  
Knock Out Mice

The retinoblastoma (RB) tumor suppressor is functionally inactivated in a wide range of human tumors where this inactivation promotes tumorigenesis in part by allowing uncontrolled proliferation. RB has been extensively studied, but its mechanisms of action in normal and cancer cells remain only partly understood. Here, we describe a new mouse model to investigate the consequences of RB depletion and its re-activation in vivo. In these mice, induction of shRNA molecules targeting RB for knock-down results in the development of phenotypes similar to Rb knock-out mice, including the development of pituitary and thyroid tumors. Re-expression of RB leads to cell cycle arrest in cancer cells and repression of transcriptional programs driven by E2F activity. Thus, continuous RB loss is required for the maintenance of tumor phenotypes initiated by loss of RB, and this new mouse model will provide a new platform to investigate RB function in vivo.

scholarly journals Interleukin (IL)-10 Is an Effective Therapeutic for Murine Transfusion Related Acute Lung Injury (TRALI)

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 92-92
Author(s):  
Rick Kapur ◽  
Michael Kim ◽  
Shanjee Shanmugabhavananthan ◽  
Jonathan Liu ◽  
Noel Kim ◽  
...  
Keyword(s):  
T Cells ◽  
Acute Lung Injury ◽  
Lung Injury ◽  
Pulmonary Edema ◽  
Mhc Class I ◽  
Lung Tissue ◽  
Cd4 T Cells ◽  
Class I ◽  
Second Hit ◽  
Tissue Histology

Abstract Transfusion related acute lung injury (TRALI) is the leading cause of transfusion-induced fatalities and is characterized by acute respiratory distress following blood transfusion. Donor antibodies present in the transfused blood product such as anti-human leukocyte antigen (HLA) or anti-human neutrophil antigen (HNA) antibodies are frequently involved. Currently, there is no treatment available for TRALI apart from supportive measures such as oxygen. The pathogenesis the disorder is incompletely understood, however, several animal models have contributed to our understanding of TRALI disease pathology. Most TRALI reactions are considered to be due to a two-hit paradigm where the first hit is a predisposing patient factor such as inflammation while the second hit is the transfusion. It is widely believed that the second hit delivers antibodies that trigger TRALI in the recipient. The anti-MHC class I antibody, 34-1-2s, has been widely used as an agent that delivers the second TRALI hit in mice. We have previously shown that CD4+ T cells, more specifically, CD4+CD25+FoxP3+ T-regulatory cells (Tregs) convey protection against TRALI (Blood. 126 (23):2342, 2015; abstract #82075, manuscript submitted). In the current study, we utilized a C57BL/6 mouse model of severe TRALI by first depleting mice of CD4+ T cells and then injecting them with the anti-MHC class I monoclonal antibodies (34-1-2s+AF6-88.5.5.3) and we examined the effects of the anti-inflammatory cytokine IL-10 on the antibody-mediated TRALI reaction. IL-10 (45 µg/kg iv) or volume-matched PBS was injected 15 minutes after the administration of anti-MHC antibodies when the onset of TRALI symptoms (e.g. a 2 degree drop in rectal temperature indicative of systemic shock) began. Results show that 90 minutes after anti-MHC class I antibody injection, control mice injected with PBS exhibited a high degree of pulmonary edema as assessed by significantly elevated lung wet-to-dry weight ratios (W/D: 5.84 ± 1.02). Pulmonary neutrophil levels were also found to be increased and lung tissue histology confirmed severe signs of acute lung injury. In contrast, mice injected with IL-10 completely recovered from TRALI; after 90 minutes post-antibody injection they displayed no signs of pulmonary edema (W/D: 4.76 ± 0.04, ** p<0.004 compared to mice injected with PBS) and no signs of severe acute lung injury as assessed by lung tissue histology. Pulmonary neutrophil levels, however, were equally increased in both groups indicating that although IL-10 rescues the mice from acute lung injury, it does not interfere with pulmonary neutrophil recruitment. Preliminary data suggests that IL-10 administration interferes with the ability of neutrophils to generate reactive oxygen species (ROS) that mediate lung injury. Our results suggest that IL-10 therapy significantly rescues an ongoing severe TRALI reaction and this may prove to be an effective and feasible therapeutic strategy in combating human TRALI. Disclosures No relevant conflicts of interest to declare.

scholarly journals MHC class I–specific antibody binding to nonhematopoietic cells drives complement activation to induce transfusion-related acute lung injury in mice

2011 ◽  
Vol 208 (12) ◽  
pp. 2525-2544 ◽  
Author(s):  
Richard T. Strait ◽  
Wyenona Hicks ◽  
Nathaniel Barasa ◽  
Ashley Mahler ◽  
Marat Khodoun ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Acute Lung Injury ◽  
Lung Injury ◽  
Mouse Model ◽  
Mhc Class I ◽  
Vascular Endothelium ◽  
Reactive Oxygen ◽  
Class I ◽  
Oxygen Species ◽  
Pulmonary Vascular Endothelium

Transfusion-related acute lung injury (TRALI), a form of noncardiogenic pulmonary edema that develops during or within 6 h after a blood transfusion, is the most frequent cause of transfusion-associated death in the United States. Because development of TRALI is associated with donor antibodies (Abs) reactive with recipient major histocompatibility complex (MHC), a mouse model has been studied in which TRALI-like disease is caused by injecting mice with anti–MHC class I monoclonal Ab (mAb). Previous publications with this model have concluded that disease is caused by FcR-dependent activation of neutrophils and platelets, with production of reactive oxygen species that damage pulmonary vascular endothelium. In this study, we confirm the role of reactive oxygen species in the pathogenesis of this mouse model of TRALI and show ultrastructural evidence of pulmonary vascular injury within 5 min of anti–MHC class I mAb injection. However, we demonstrate that disease induction in this model involves macrophages rather than neutrophils or platelets, activation of complement and production of C5a rather than activation of FcγRI, FcγRIII, or FcγRIV, and binding of anti–MHC class I mAb to non-BM–derived cells such as pulmonary vascular endothelium. These observations have important implications for the prevention and treatment of TRALI.

scholarly journals Mechanical ventilation aggravates transfusion-related acute lung injury induced by MHC class I antibodies

Critical Care ◽  
2010 ◽  
Vol 14 (Suppl 1) ◽  
pp. P192
Author(s):  
AP Vlaar ◽  
EK Wolthuis ◽  
JJ Hofstra ◽  
JJ Roelofs ◽  
L Boon ◽  
...  
Keyword(s):  
Mechanical Ventilation ◽  
Acute Lung Injury ◽  
Lung Injury ◽  
Mhc Class I ◽  
Class I
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