Methods for controlling adverse events in inflammatory patients

Abstract

The present invention relates to methods for controlling adverse events in inflammatory patients, methods for preventing and reversing inflammation.

Description

[0001] This application is a divisional application of a Chinese patent application with an application date of November 14, 2019, an application number of 201980079192.0, and an invention title of "Methods for Controlling Adverse Events in Inflammatory Patients".

[0002] Cross - reference to related applications

[0003] This application claims priority to U.S. Provisional Patent Application Serial No. 62 / 768,667, filed on November 16, 2018.

[0004] Government rights

[0005] This invention was made with government support under National Heart, Lung, and Blood Institute grant number R44HL132172. The government has certain rights in the invention. Technical field

[0006] This disclosure relates to the treatment of inflammation, sickle cell disease, trauma, and hemorrhagic shock. Background art

[0007] Sickle cell disease (SCD), a group of inherited red blood cell disorders, affects millions of people worldwide. It is estimated that SCD affects approximately 100,000 Americans.

[0008] Sickle cell disease is divided into six classes: sickle cell anemia (HbSS), hemoglobin C - sickle cell anemia (HbSC), hemoglobin S - beta - thalassemia ((HbSβ0 and HbSβ+ thalassemia), hemoglobin SD disease (HbSD), hemoglobin SE disease (HbSE), and hemoglobin SO Arab disease (HbSO). The most severe form, HbSS, is characterized by the inheritance of two sickle cell genes. In HbSC, a single sickle cell gene is inherited from one parent, and an abnormal gene for hemoglobin C is inherited from the other parent. HbSC is a milder form of the disease. People with HbSβ thalassemia inherit one sickle cell gene and one beta - thalassemia gene. HbSβ thalassemia is further divided into beta - thalassemia O and beta - thalassemia +, with beta - thalassemia O being the more severe of the two. HbSD, HbSE, and HbSO are rare types of sickle cell disease. In the rare types, a sickle cell gene and a gene from an abnormal hemoglobin type (D, E, or O) are inherited.

[0009] Some of the most common complications of SCD include vaso - occlusion. Studies have shown that sickle RBCs (SS - RBCs) initiate vaso - occlusive episodes through adhesive interactions with the endothelium. Vaso - occlusive episodes cause pain of varying degrees, which varies from person to person. This pain can start suddenly and last for any period of time. Patients are also more likely to experience infections, including influenza, meningitis, and hepatitis. Patients may also experience hand - foot syndrome, eye diseases, acute chest syndrome, and stroke. In addition, studies have shown that sickle cell disease promotes an inflammatory response. See Plat, O., “Sickle cell anemia as an inflammatory disease” J. Clin. Invest., 106(3):337 - 338 (2000). SCD can damage a patient's brain, eyes, heart, lungs, liver, spleen, kidneys, skin, joints, and bones throughout their life.

[0010] Treatment for sickle cell disease includes blood transfusions, bone marrow transplantation, stem cell transplantation, and medications (such as hydroxyurea) that reduce the number of pain episodes during crisis periods.

[0011] In 2010, 5.1 million people died as a result of injuries, exceeding the number of deaths due to the combined effects of HIV, tuberculosis, and malaria (3.8 million). See Norton et al., “Global Health Injuries,” The New England Journal of Medicine (The NEJM) 368:1723 - 30 (2013) (“Norton 2013”) (hereby incorporated by reference in its entirety). Injuries include unintentional injuries (e.g., road traffic accidents, falls, and burns) and intentional injuries (e.g., self - harm, interpersonal violence, war, and conflict). See Norton 2013. Between 1990 and 2010, the number of injury - related deaths worldwide increased by 24%, but in the United States between 2000 and 2010, the number of injury - related deaths increased by 23%. See Norton 2013. Additionally, at least 20% of all trauma deaths are the result of potentially survivable injuries and can therefore be prevented through optimal care. Fox et al., “Earlier Endpoints are Required for Hemorrhagic Shock Trials Among Severely Injured Patients.” Shock, 47:567 - 73 (2017) (hereby incorporated by reference in its entirety). The percentage of preventable deaths makes it necessary to develop therapies for avoidable complications that lead to death.

[0012] Penetrating injuries (e.g., gunshot or stab wounds) and blunt trauma (e.g., falls or motor vehicle accidents) are the major causes of hemorrhagic trauma. The resulting shock is a condition of insufficient oxygen supply to tissues caused by massive bleeding, leading to oxygen debt, anaerobic metabolism, and elevated plasma lactate levels. Shock that cannot be reversed by restoring circulation and oxygen delivery may result in permanent tissue damage, multiple organ failure, and death.

[0013] Clinical sequelae of hemorrhagic trauma and shock include death due to exsanguination within hours of trauma, as well as death after 24 hours of morbidity from trauma and massive transfusion. Such morbidity includes multiple organ failure involving the lungs, kidneys, and liver caused by acute traumatic coagulopathy or inflammation, and infection / sepsis caused by transfusion-related immunomodulation; both morbidities are increased by the lower quality of the blood products transfused and the higher volume of pRBCs transfused.

[0014] One method of treating hemorrhagic shock is resuscitation with crystalloids. However, the use of crystalloids leads to increased morbidity and mortality by causing trauma-induced coagulopathy. At least for this reason, early administration of blood components is advocated to reverse shock caused by hemorrhagic trauma. Packed red blood cells (pRBCs) are transfused into patients with hemorrhagic trauma to restore the lost blood volume, restore the patient's oxygen-carrying capacity, and restore oxidative metabolism in tissues from anaerobic metabolism. However, the use of pRBCs is not without the risk of complications, including antigen mismatch, pathogen transmission, circulatory overload, and degradation of pRBCs during storage ex vivo.

[0015] When stored conventionally, stored blood undergoes a steady deterioration, which is associated with various storage lesions, including hemolysis, hemoglobin degradation, and decreased ATP and 2,3-DPG concentrations. When transfused into patients, the effects of the steady deterioration during storage are manifested as, for example, reduced in vivo recovery at 24 hours. The rapid decrease in hematocrit is due to the reduced 24-hour recovery and, in severe cases, can lead to delayed hemolytic transfusion reactions (DHTRs). Other complications, such as systemic inflammatory response syndrome (SIRS), transfusion-related acute lung injury (TRALI), and transfusion-related immunomodulation (TRIM), are associated with the transfusion of stored blood, although the identification of the underlying causes remains unclear.

[0016] Even when transfused within the current six-week limit, stored RBCs tend to exhibit lower quality (e.g., increased proportion of removed RBCs; impaired oxygen exchange capacity; reduced deformability) and increased toxicity, which typically manifests as clinical sequelae of transfusion therapy. A large and growing body of literature in the literature supports this view. See Zimring, "Established and theoretical factors to consider in assessing the red cell storage lesion," Blood, 125:2185-90 (2015); Zhu et al., "Impaired adenosine-5'-triphosphate release from red blood cells promotes their adhesion to endothelial cells: a mechanism of hypoxemia after transfusion," Critical care medicine, 39:2478-86 (2011); Weinberg et al., "Red blood cell age and potentiation of transfusion-related pathology in trauma patients," Transfusion, 51:867-73 (2011); Spinella et al., "Does the storage duration of blood products affect outcomes in critically ill patients?" Transfusion 51:1644-50 (2011); Roback et al., "Insufficient nitric oxide bioavailability: a hypothesis to explain adverse effects of red blood cell transfusion," Transfusion, 51:859-66 (2011);Reynolds et al., "The transfusion problem: role of aberrant S-nitrosylation," Transfusion, 51:852-8 (2011); Kim-Shapiro et al., "Storage lesion: role of red blood cell breakdown," Transfusion, 51:844-51 (2011); Jy et al., "Microparticles in stored red blood cells as potential mediators of transfusion complications," Transfusion, 51:886-93 (2011); Hod et al., "Transfusion of human volunteers with older, stored red blood cells produces extravascular hemolysis and circulating non-transferrin-bound iron," Blood, 118:6675-82 (2011); Flegel et al., "Does prolonged storage of red blood cells cause harm?" British journal of haematology 165:3-16 (2014);Redlin et al., "Red blood cell storage duration is associated with various clinical outcomes in pediatric cardiac surgery," Transfusion medicine and hemotherapy: offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie 41:146-51 (2014); Rogers et al., "Storage duration of red blood cell transfusion and Clostridium difficile infection: a within person comparison," PLoS One 9:e89332 (2014); Spinella et al., "Properties of stored red blood cells: understanding immune and vascular reactivity," Transfusion 51:894-900 (2011); Brown et al., "Length of red cell unit storage and risk for delirium after cardiac surgery," Anesth Analg, 119:242-50 (2014); Wang et al., "Transfusion of older stored blood worsens outcomes in canines depending on the presence and severity of pneumonia," Transfusion, 54:1712-24 (2014);Liu et al., "Mechanism of faster NO scavenging by older stored red blood cells," Redox biology, 2:211-9 (2014); Prestia et al., "Transfusion of stored blood impairs host defenses against Gram-negative pathogens in mice," Transfusion 54:2842-51 (2014); D'Alessandro et al., "An update on red blood cell storage lesions, as gleaned through biochemistry and omics technologies," Transfusion, 55:205-19 (2015) (hereby incorporated by reference in its entirety). A large number of in vitro studies have clearly shown the degradation of RBCs (storage lesions) during conventional storage. A large number of emerging metabolomics studies have shown the development of storage lesions at the molecular level. See Roback et al., "Metabolomics of AS-1 RBCs Storage," Transfusion medicine reviews (2014); D'Alessandro et al., "Metabolomics of AS-5 RBCs supernatants following routine storage," Vox sanguinis (2014); D'Alessandro et al., "Routine storage of red blood cell (RBC) units in additive solution-3: a comprehensive investigation of the RBC metabolome," Transfusion 55:1155-68 (2015);D'Alessandro et al., "Red blood cell storage in additive solution-7 preserves energy and redox metabolism: a metabolomics approach," Transfusion (2015); Wither et al., "Hemoglobin oxidation at functional amino acid residues during routine storage of red blood cells," Transfusion (2015); D'Alessandro et al., "Citrate metabolism in red blood cells stored in additive solution-3," Transfusion (2016); D'Alessandro et al., "Omics markers of the red cell storage lesion and metabolic linkage," Transfusion, 15:137-44 (2017) (hereby incorporated by reference in its entirety). It is necessary to reduce or prevent such degradation to improve the efficacy of transfusion (more O2 is immediately delivered to peripheral tissues after transfusion) and reduce mortality due to inflammation, sickle cell disease, or hemorrhagic trauma.;

[0017] Oxidative damage triggers multiple RBC storage lesions and their downstream consequences in conventionally stored blood; thus, methods to reduce the degree of oxidative stress are needed to reduce RBC storage lesions. A variety of methods have been developed to minimize storage lesions and improve transfusion outcomes. Methods include additive solutions (e.g., U.S. Patent No. 4,769,318 to Hamasaki et al., U.S. Patent No. 4,880,786 to Sasakawa et al., and U.S. Patent No. 6,447,987 to Hess et al.), cryopreservation (see U.S. Patent No. 6,413,713 to Serebrennikov Chaplin et al., "Blood Cells for Transfusion," Blood, 59:1118-20 (1982), and Valeri et al., "The survival, function, and hemolysis of human RBCs stored at 4 degrees C in additive solution (AS-1, AS-3, or AS-5) for 42 days and then biochemically modified, frozen, thawed, washed, and stored at 4 degrees C in sodium chloride and glucose solution for 24 hours," Transfusion, 40:1341-5 (2000)) (which is hereby incorporated by reference in its entirety).

[0018] One method that has proven successful in improving blood quality and extending its utility is by depleting oxygen and storing it under anaerobic conditions. The benefits of storing blood under oxygen-depleted conditions are increased levels of ATP and 2,3-DPG and reduced hemolysis. U.S. Patent No. 5,624,794 to Bitensky et al., U.S. Patent No. 6,162,396 to Bitensky et al., and U.S. Patent No. 5,476,764 to Bitensky (each incorporated herein by reference in its entirety) relate to the storage of red blood cells under oxygen-depleted conditions. U.S. Patent No. 5,789,151 to Bitensky et al. (incorporated herein by reference in its entirety) relates to a blood storage additive solution. U.S. Patent No. 6,162,396 ('396 patent) to Bitensky et al. (incorporated herein by reference in its entirety) discloses an anaerobic storage bag for blood storage, which includes an oxygen-impermeable outer layer, an oxygen-permeable red blood cell (RBC)-compatible inner layer, and an oxygen scrubber disposed between the inner and outer layers.

[0019] Compared to blood stored under conventional conditions, storing blood under oxygen-depleted conditions also results in reduced levels of microparticles, reduced loss of deformability, reduced lipid and protein oxidation, and higher post-transfusion survival. See Yoshida et al., "The effects of additive solution pH and metabolic rejuvenation on anaerobic storage of red cells," Transfusion 48:2096-2105 (2008) and Yoshida, T., et al. "Reduction of microparticle generation during anaerobic storage of red blood cells," Transfusion, 52, 83A (2012) (which are hereby incorporated by reference in their entirety). Anaerobically stored RBCs also provide higher 24-hour in vivo recovery after autologous transfusion, higher 2,3-DPG and ATP levels, lower hemolysis, and beneficial metabolic pathway remodeling. See Reisz et al "Oxidative modifications of glyceraldehyde 3-phosphate dehydrogenase regulate metabolic reprogramming of stored red blood cells," Blood, 128:e32-42 (2016); and Yoshida et al., "Extended storage of red blood cells under anaerobic conditions," Blood Voices 92:22-31 (2007) (which are hereby incorporated by reference in their entirety).

[0020] In the present disclosure, we demonstrate that oxygen-reduced (OR) or oxygen- and carbon dioxide-reduced (OCR) red blood cells can reduce vascular occlusion in SCD patients by reducing adhesion to thrombospondin. We also demonstrate that OR and OCR red blood cells have reduced hemolysis in the presence of plasma from SCD patients compared to conventionally stored blood.

[0021] In the present disclosure, we demonstrate that compared to conventionally stored blood, OR or OCR blood from rats provides improved ATP and 2,3-DPG during storage, as previously demonstrated using human blood. Thus, OR or OCR rat RBCs are expected to have similar reductions in microparticles, improved deformability, reduced lipid and protein oxidation, and higher post-transfusion survival.

[0022] Here we demonstrate for the first time that when infused to treat hemorrhagic trauma, OR and OCR blood in rats provides remarkable improvements in clinical outcomes. Using a rat hemorrhagic shock resuscitation model, it is shown that OR or OCR RBCs provide reduced organ injury relative to conventionally stored blood. In addition, OR or OCR RBCs provide reversal of the shock state using a smaller pRBC volume. Finally, when infused to treat hemorrhagic shock, OR or OCR RBCs more rapidly stabilize hemodynamics relative to pRBCs of the same storage duration stored conventionally.

[0023] OR and OCR RBCs provide an improved method for treating trauma resulting in blood loss to reduce mortality and morbidity relative to conventionally stored blood. OR and OCR RBCs provide reduced organ failure, including reduced levels of markers of lung and liver injury. OR and OCR RBCs further reduce the volume of blood required to restore and stabilize hemodynamic function. Thus, OR and OCR RBCs can reduce the RBC volume required for transfusion therapy when treating hemorrhagic trauma. The improved quality of OR and OCR, in addition to the previously demonstrated improvement in the oxygen delivery capacity of stored RBCs, provides an unexpected reduction in trauma-related organ injury, morbidity, and mortality. SUMMARY OF THE INVENTION

[0024] The present disclosure provides and includes a method of treating a patient in need thereof with stored oxygen-reduced blood having an oxygen saturation of 20% or less during storage, wherein the patient in need thereof has an inflammation.

[0025] The present disclosure provides and includes a method of improving transfusion in a patient in need thereof, the method comprising providing stored oxygen-reduced blood to a patient having sickle cell disease, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage.

[0026] The present disclosure provides and includes a method of reducing the number of vaso-occlusive episodes in a patient in need thereof, the method comprising providing stored oxygen-reduced blood to a patient in need thereof, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage, and wherein reducing the vaso-occlusive episodes comprises reducing the adhesion of red blood cells to endothelial cells.

[0027] The present disclosure provides and includes a method of reducing the adhesion of red blood cells to endothelial cells in a patient in need thereof, the method comprising providing stored oxygen-reduced blood to a patient suffering from sickle cell disease, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The present disclosure is provided with reference to the accompanying drawings, in which:

[0029] Figure 1 is a graph presenting the results according to an exemplary embodiment of the present disclosure, comparing the ATP levels in conventionally stored RBCs (untreated; control), sham control (SC), oxygen-reduced RBCs (N2; OR), and oxygen- and carbon dioxide-reduced RBCs (CO2; OCR).

[0030] Figure 2 is a graph presenting the results according to an exemplary embodiment of the present disclosure, comparing the 2,3-DPG levels in conventionally stored RBCs (untreated; control), sham control (SC), oxygen-reduced RBCs (N2; OR), and oxygen- and carbon dioxide-reduced RBCs (CO2; OCR).

[0031] Figure 3 is a graph presenting the results according to an exemplary embodiment of the present disclosure, presenting a comparison of the recovery percentages of control, sham, OR-RBC, and OCR-RBC infused into animals.

[0032] Figure 4A and 4B is a graph presenting the results according to an exemplary embodiment of the present disclosure, presenting a comparison of the hematocrit percentages in animals resuscitated with control, OR-RBC, and OCR-RBC stored for 1 week ( Figure 4A ) or 3 weeks ( Figure 4B ). BL (baseline) represents animals not under shock conditions. Shock represents animals under hemorrhagic shock. Early R represents a 10-minute resuscitation period. Late R represents a 60-minute resuscitation period.

[0033] Figure 5A and 5B is a graph presenting the results according to an exemplary embodiment of the present disclosure, providing a comparison of the mean arterial pressure (MAP) in animals resuscitated with control, OR-RBC, and OCR-RBC stored for 1 week ( Figure 5A ) or 3 weeks ( Figure 5B ). BL (baseline) represents animals not under shock conditions. Shock represents animals under hemorrhagic shock. Early R represents a 10-minute resuscitation period. Late R represents a 60-minute resuscitation period.

[0034] Figure 6A and6B is a graph presenting the results of an exemplary embodiment according to the present disclosure, providing a comparison of the percentage of blood volume provided to an animal during resuscitation after 10, 20, 30, 45, and 60 minutes. Controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 6A ) or 3 weeks ( Figure 6B ) were compared.

[0035] Figure 7A and 7B is a graph presenting the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of lactic acid in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 7A ) or 3 weeks ( Figure 7B ). BL (baseline) represents animals not under shock conditions. Shock represents animals under hemorrhagic shock. Early R represents a 10-minute resuscitation period. Late R represents a 60-minute resuscitation period.

[0036] Figure 8A and 8B is a graph presenting the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of glucose in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 8A ) or 3 weeks ( Figure 8B ). BL (baseline) represents animals not under shock conditions. Shock represents animals under hemorrhagic shock. Early R represents a 10-minute resuscitation period. Late R represents a 60-minute resuscitation period.

[0037] Figure 9A and 9B is a graph presenting the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of AST in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 9A ) or 3 weeks ( Figure 9B ).

[0038] Figure 10A and 10B is a graph presenting the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of ALT in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 10A ) or 3 weeks ( Figure 10B ).

[0039] Figure 11A and 11B is a graph presenting the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of serum creatinine in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 11A ) or 3 weeks ( Figure 11B ).

[0040] Figure 12A and 12B are graphs presenting the results according to exemplary embodiments of the present disclosure, providing a comparison of the amount of blood urea nitrogen (BUN) in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 12A ) or 3 weeks ( Figure 12B ).

[0041] Figure 13A and 13B are graphs presenting the results according to exemplary embodiments of the present disclosure, providing a comparison of the amount of CXCL1 in the livers of animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 13A ) or 3 weeks ( Figure 13B ).

[0042] Figure 14A and 14B are graphs presenting the results according to exemplary embodiments of the present disclosure, providing a comparison of the amount of CXCL1 in the spleens of animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 14A ) or 3 weeks ( Figure 14B ).

[0043] Figure 15A and 15B are graphs presenting the results according to exemplary embodiments of the present disclosure, providing a comparison of the amount of CXCL1 in the lungs of animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 15A ) or 3 weeks ( Figure 15B ).

[0044] Figure 16A and 16B are graphs presenting the results according to exemplary embodiments of the present disclosure, providing a comparison of the amount of urinary neutrophil gelatinase-associated lipocalin (u-NGAL) in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 16A ) or 3 weeks ( Figure 16B ).

[0045] Figure 17A and 17B are graphs presenting the results according to exemplary embodiments of the present disclosure, providing a comparison of the percentage of CD45+ neutrophils in animals resuscitated with controls, OR-RBCs, and OCR-RBCs stored for 1 week ( Figure 17A ) or 3 weeks ( Figure 17B ).

[0046] Figure 18A and 18Bis a graph presenting the results of exemplary embodiments according to the present disclosure, providing a comparison of the amount of IL-6 in animals resuscitated with control, OR-RBC, and OCR-RBC stored for 1 week ( Figure 18A ) or 3 weeks ( Figure 18B ).

[0047] Figures 19A to 19C is a graph presenting the results of exemplary embodiments according to the present disclosure, providing a comparison of the adhesion of red blood cells to thrombospondin.

[0048] Figures 20A to 20C is a graph presenting the results of exemplary embodiments according to the present disclosure, providing a comparison of the adhesion of red blood cells to thrombospondin between conventionally stored or oxygen-reduced stored red blood cells.

[0049] Figure 21 shows the results of exemplary embodiments according to the present disclosure, providing a comparison of hemolysis in conventionally stored red blood cells, oxygen-reduced stored red blood cells, and oxygen- and carbon dioxide-reduced stored red blood cells incubated with healthy sickle cell plasma.

[0050] The examples set forth herein illustrate several embodiments of the present disclosure, but should not be construed as limiting the scope of the present disclosure in any way. DETAILED DESCRIPTION

[0051] The methods of the present disclosure provide and include providing to a hemorrhagic patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. The methods also provide providing to a hemorrhagic patient oxygen-reduced stored blood having an oxygen saturation of 15% to 20% before and during storage. The methods also provide providing to a hemorrhagic patient oxygen-reduced stored blood having an oxygen saturation of 10% to 15% before and during storage. The methods also provide providing to a hemorrhagic patient oxygen-reduced stored blood having an oxygen saturation of 5% to 10% before and during storage. The methods also provide providing to a hemorrhagic patient oxygen-reduced stored blood having an oxygen saturation of 3% to 5% before and during storage.

[0052] The method also provides oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage for transfusion to a person suffering from hemorrhagic shock. The method also provides oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage for transfusion to a person suffering from hemorrhagic trauma. The method also includes transfusing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient having an increased risk of trauma due to surgery. The method of providing oxygen-reduced stored blood having an initial oxygen saturation of 20% or less includes providing oxygen-reduced stored blood having an initial oxygen saturation of 10% or less. The method of providing oxygen-reduced stored blood having an initial oxygen saturation of 20% or less also includes providing oxygen-reduced stored blood having an initial oxygen saturation of 5% or less. The method of providing oxygen-reduced stored blood having an initial oxygen saturation of 20% or less also includes providing oxygen-reduced stored blood having an initial oxygen saturation of 3% or less.

[0053] The method of the present disclosure provides and includes providing stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 20% or less before and during a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 15% or less after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 10% or less after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method further provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 5% or less after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method further provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 3% or less after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 3% to 5% after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 5% to 10% after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 10% to 15% after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides stored blood with reduced oxygen for treating trauma, the blood having an oxygen saturation of 15% to 20% after a storage period of at least one week, at least two weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks.

[0054] The method of the present disclosure provides and includes providing a trauma patient with stored blood with reduced oxygen having an oxygen saturation of 20% or less before and during storage. In one aspect, the trauma patient suffers from a head trauma, a penetrating wound, a blunt force trauma, a fall injury, or a car accident injury. In another aspect, the trauma patient is a hemorrhagic trauma patient. In yet another aspect, the trauma patient bleeds due to surgery, a penetrating wound, a blunt force trauma, a fall injury, or a car accident injury.

[0055] In one aspect of the present disclosure, a trauma patient or a hemorrhagic trauma patient is a subject in need of blood stored in OR and OCR. In aspects of the present disclosure, a trauma patient is a hemorrhagic trauma patient in need of one or more blood units for transfusion therapy. In aspects of the present disclosure, a trauma patient is a hemorrhagic trauma patient in need of two or more blood units for transfusion therapy. In aspects of the present disclosure, a trauma patient is a hemorrhagic trauma patient in need of three or more blood units for transfusion therapy.

[0056] In one aspect of the present disclosure, a trauma patient is a patient in hemorrhagic shock. In one aspect, the trauma patient is in hemorrhagic shock due to head trauma, penetrating trauma, blunt force trauma, fall injury, or motor vehicle crash injury. In aspects of the present disclosure, a hemorrhagic trauma patient is a patient with Class I hemorrhage. In another aspect, a hemorrhagic trauma patient is a patient with Class II hemorrhage. In another aspect, a hemorrhagic trauma patient is a patient with Class III hemorrhage. In another aspect, a hemorrhagic trauma patient is a patient with Class IV hemorrhage. In one aspect of the present disclosure, a hemorrhagic trauma patient loses up to 15% of their blood volume. In another aspect, a hemorrhagic trauma patient loses 15 to 30% of their blood volume. In another aspect, a hemorrhagic trauma patient loses 30 to 40% of their blood volume. In another aspect, a hemorrhagic trauma patient loses more than 40% of their blood volume.

[0057] The present disclosure provides and includes that patients in need of blood transfusion therapy with OR or OCR RBC exhibit one or more signs selected from the group consisting of decreased mean arterial pressure, decreased hematocrit, increased lactate, increased glucose, increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL), increased serum creatinine, and increased blood urea nitrogen. In one aspect of the present disclosure, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with decreased mean arterial pressure. The present disclosure provides and includes that patients in need of blood transfusion therapy with OR or OCR RBC exhibit increased aspartate aminotransferase (AST) and increased alanine aminotransferase (ALT). The present disclosure provides and includes that patients in need of blood transfusion therapy with OR or OCR RBC exhibit decreased mean arterial pressure and increased lactate. The present disclosure provides and includes that patients in need of blood transfusion therapy with OR or OCR RBC exhibit increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), and increased blood urea nitrogen. The present disclosure provides and includes that patients in need of blood transfusion therapy with OR or OCR RBC exhibit increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), increased serum creatinine, and increased blood urea nitrogen. The present disclosure provides and includes that patients in need of blood transfusion therapy with OR or OCR RBC exhibit increased lactate and increased glucose. The present disclosure provides and includes that patients in need of blood transfusion therapy with OR or OCR RBC exhibit increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL), increased serum creatinine, and increased blood urea nitrogen.

[0058] In another aspect, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with decreased hematocrit. In another aspect, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with increased lactate. In yet another aspect, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with increased glucose. In a further aspect, the hemorrhagic trauma patient has increased aspartate aminotransferase (AST). In another aspect, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with increased alanine aminotransferase (ALT). In another aspect, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL). In another aspect, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with increased serum creatinine. In another aspect, the patient in need of blood transfusion therapy with OR or OCR RBC is a hemorrhagic trauma patient with increased blood urea nitrogen.

[0059] In one aspect of the present disclosure, the blood stored for OR and OCR for transfusion therapy for a patient in need has an initial oxygen saturation of 20% or less. In another aspect, the blood stored for OR and OCR has an initial oxygen saturation of 10% or less. In another aspect, the blood stored for OR and OCR has an initial oxygen saturation of 5% or less. In another aspect, the blood stored for OR and OCR has an initial oxygen saturation of 3% or less.

[0060] In one aspect of the present disclosure, the blood stored for OCR for transfusion therapy for a patient in need has an initial pCO2 (at 37 °C) of 10 to 40 mmHg. In another aspect, the blood stored for OCR has an initial pCO2 of 10 to 30 mmHg. In another aspect, the blood stored for OCR has an initial pCO2 of 10 to 20 mmHg. In another aspect, the blood stored for OCR has an initial pCO2 of 10 to 15 mmHg. In yet another aspect, the blood stored for OCR has an initial pCO2 of less than 10 mmHg.

[0061] In one aspect of the present disclosure, the blood stored for OR and OCR for transfusion therapy for a patient in need has an initial oxygen saturation of 20% or less and is stored for less than 2 days. In one aspect, the blood stored for OR and OCR has an initial oxygen saturation of 20% or less and is stored for less than 7 days. In another aspect, the blood stored for OR and OCR has an initial oxygen saturation of 20% or less and is stored for less than 14 days. In another aspect, the blood stored for OR and OCR has an initial oxygen saturation of 20% or less and is stored for less than 21 days. In another aspect, the oxygen-reduced stored blood for transfusion therapy for a patient in need has an initial oxygen saturation of 20% or less and is stored for less than 28 days. In another aspect, the oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 35 days. In another aspect, the oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 42 days. In another aspect, the oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 45 days. In one aspect of the present disclosure, the blood stored for OR and OCR has an initial oxygen saturation of 20% or less during storage.

[0062] Suitable blood for transfusion therapy for a patient in need includes oxygen-reduced stored blood with an anticoagulant. In one aspect of the present disclosure, oxygen-reduced red blood cells are stored for up to 3 weeks to produce oxygen-reduced stored blood. In another aspect, the oxygen-reduced stored blood generally further comprises an additive solution. Suitable additive solutions according to the present disclosure include AS-1, AS-3 AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, OFAS3, and combinations thereof. In one aspect, an additive solution is added during component separation. In one aspect, the additive solution is AS-1, and in another aspect, the additive solution is AS-3. In other aspects, the additive solution is SAGM.

[0063] The methods of the present disclosure provide for and include increasing the mean arterial pressure (MAP) in hemorrhagic trauma patients in need of blood transfusion therapy, including providing to a trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the mean arterial pressure is increased by 20% to 60%. In another aspect, the mean arterial pressure is increased by 30% to 60%. In another aspect, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased by 30% to 50%. In yet another aspect, the mean arterial pressure is increased by 30% to 60%. In a further aspect, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased by 30% to 40%. In one aspect, the mean arterial pressure is increased by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% compared to the mean arterial pressure of a patient receiving conventional stored blood.

[0064] In one aspect of the present disclosure, the mean arterial pressure of a patient receiving transfusion therapy with OR or OCR blood is increased by at least 1.5-fold. In another aspect, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased by at least 2-fold. In a further aspect, the mean arterial pressure of a patient receiving transfusion therapy with OR or OCR blood is increased by 1 to 2-fold. In one aspect of the present disclosure, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased by at least 10 mmHg, at least 20 mmHg, at least 30 mmHg, at least 40 mmHg, at least 50 mmHg, or at least 60 mmHg. In another aspect, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased by 20 mmHg to 50 mmHg. In a further aspect, the mean arterial pressure of a patient receiving transfusion therapy with OR or OCR blood is increased by 30 mmHg to 50 mmHg.

[0065] The methods of the present disclosure provide and include increasing the mean arterial pressure of a trauma patient in need of blood transfusion therapy to 70 mmHg to 110 mmHg, including providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In another aspect, the mean arterial pressure of a trauma patient receiving a blood transfusion therapy with OR or OCR blood is increased to at least 70 mmHg. In another aspect, the mean arterial pressure of a trauma patient receiving a blood transfusion therapy with OR or OCR blood is increased to at least 80 mmHg. In yet another aspect, the mean arterial pressure is increased to at least 90 mmHg. In a further aspect, the mean arterial pressure is increased to at least 100 mmHg. In one aspect of the present disclosure, the mean arterial pressure of a subject in need remains at 70 mmHg to 110 mmHg for at least 1 hour after blood transfusion. In another aspect, the mean arterial pressure remains at 70 mmHg to 110 mmHg for at least 2 hours after blood transfusion. In yet another aspect, the mean arterial pressure remains at 70 mmHg to 105 mmHg for at least 3 hours after blood transfusion. In another aspect, the mean arterial pressure remains at 70 mmHg to 110 mmHg for at least 4 hours after blood transfusion. In another aspect, the mean arterial pressure remains at 70 mmHg to 110 mmHg for at least 5 hours after blood transfusion.

[0066] The methods of the present disclosure provide and include increasing the mean arterial pressure of a trauma patient in need of blood transfusion therapy at a faster rate than the mean arterial pressure of a patient infused with conventional stored blood, including providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the mean arterial pressure of a patient infused with OR or OCR blood returns to within the normal physiological parameter range in half the time compared to conventional stored blood.

[0067] The methods of the present disclosure provide and include reducing the amount of stored blood required for transfusion in a hemorrhagic trauma patient in need of blood transfusion therapy, including providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the amount of OR-stored blood required for transfusion is 10% to 90% less than the amount required for conventional stored blood. In another aspect, the amount of OR-stored blood required for transfusion is 10% to 30% less than the amount required for conventional stored blood. In another aspect, the amount of OR-stored blood required for transfusion is 20% to 50% less than the amount required for conventional stored blood. In another aspect, the amount of OR-stored blood required for transfusion is 20% to 80% less than the amount required for conventional stored blood. In another aspect, the amount of OR-stored blood required for transfusion is 30% to 80% less than the amount required for conventional stored blood. In yet another aspect, the amount of OR-stored blood required for transfusion is 40% to 85% less than the amount required for conventional stored blood. In a further aspect, the amount of OR-stored blood required for transfusion is 50% to 90% less than the amount required for conventional stored blood.

[0068] The method of the present disclosure provides and includes reducing the amount of stored blood required for transfusion in a hemorrhagic trauma patient in need of transfusion therapy by at least 10%, including providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the amount of OR stored blood required for transfusion is at least 20% less than the amount required for conventional stored blood. In another aspect, the amount of OR stored blood required for transfusion is at least 30% less than the amount required for conventional stored blood. In another aspect, the amount of OR stored blood required for transfusion is at least 40% less than the amount required for conventional stored blood. In another aspect, the amount of OR stored blood required for transfusion is at least 50% less than the amount required for conventional stored blood. In yet another aspect, the amount of OR stored blood required for transfusion is at least 60% less than the amount required for conventional stored blood. In another aspect, the amount of OR stored blood required for transfusion is at least 70% less than the amount required for conventional stored blood. In a further aspect, the amount of OR stored blood required for transfusion is about 10% to 20%, about 20% to 30%, about 30% to 40%, about 40% to 50%, about 50% to 60%, about 60% to 70%, about 70% to 80%, about 80% to 90%, or about 90% to 95% less than the amount required for conventional stored blood. In another aspect, the amount of OR stored blood required for transfusion in a patient in need of transfusion therapy is 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90%, or 90% to 95% less than the amount required for conventional stored blood.

[0069] Lactate clearance is a biomarker for hemorrhagic shock resuscitation. See Hashmi et al., “Predictors of mortality in geriatric trauma patients: a systematic review and meta-analysis,” The journal of trauma and acute care surgery, 76:894-901 (2014); Regnier et al., “Prognostic significance of blood lactate and lactate clearance in trauma patients,” Anesthesiology, 117:1276-88 (2012); and Zhang et al., “Lactate clearance is a useful biomarker for the prediction of all-cause mortality in critically ill patients: a systematic review and meta-analysis,” Critical Care Medicine, 42:2118-25 (2014) (“Zhang 2014”) (hereby incorporated by reference in its entirety). The clinical value of lactate clearance can be used to predict the outcomes of patients with septic shock and critically ill patients without overt circulatory shock. Elevated lactate is an indicator of poor clinical outcomes, and its rapid clearance is generally associated with improved outcomes in heterogeneous ICU or ED patient populations. See Zhang 2014. The lower lactate levels in animals resuscitated with OR-RBC compared to conventional RBC support the view that resuscitation with OR RBC can significantly improve patient clinical outcomes. See Figure 7A and 7B 。

[0070] The methods of the present disclosure provide and include reducing the lactate level in a patient in need of blood transfusion therapy, including providing to the patient oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the lactate level is reduced by 10% to 90%. In one aspect, transfusion of blood stored with OR reduces the lactate level in a patient in need of blood transfusion therapy by 10% to 50%. In another aspect, transfusion of blood stored with OR reduces the lactate level in a patient in need of blood transfusion therapy by 20% to 40%. In another aspect, transfusion of blood stored with OR reduces the lactate level in a patient in need of blood transfusion therapy by 50% to 90%. In another aspect, transfusion of blood stored with OR reduces the lactate level in a patient in need of blood transfusion therapy by 60% to 90%. In another aspect, transfusion of blood stored with OR reduces the lactate level in a patient in need of blood transfusion therapy by 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, or 80% to 90%. In another aspect, transfusion of blood stored with OR reduces the lactate level in a trauma patient in need of blood transfusion therapy by at least 10%. In another aspect, transfusion of blood stored with OR reduces the lactate level in a patient in need of blood transfusion therapy by at least 20%. In a further aspect, transfusion of blood stored with OR reduces the lactate level in a patient in need of blood transfusion therapy by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%.

[0071] The method of the present disclosure provides and includes reducing an elevated lactate level in a patient in need of blood transfusion therapy to from about 0.5 mmol / L to about 2.5 mmol / L, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the lactate level of a patient in need of blood transfusion therapy is reduced to from about 0.9 mmol / L to about 2 mmol / L. In one aspect, the lactate level of a patient in need of blood transfusion therapy is reduced to from about 0.9 mmol / L to about 1.7 mmol / L. In another aspect, the lactate level of a patient in need of blood transfusion therapy is reduced to from about 1.4 mmol / L to about 2.4 mmol / L. In another aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to from about 1.7 mmol / L to about 2.5 mmol / L. In yet another aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to less than about 2.5 mmol / L. In a further aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to less than about 2.0 mmol / L. In another aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to less than about 1.5 mmol / L. In another aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to less than about 1.0 mmol / L. In yet another aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to from about 0.5 mmol / L to about 1.0 mmol / L.

[0072] The method of the present disclosure provides and includes reducing an elevated lactate level in a trauma patient in need of blood transfusion therapy to from 0.5 mmol / L to 2.5 mmol / L, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the lactate level of a patient in need of blood transfusion therapy is reduced to from 0.9 mmol / L to 2 mmol / L. In one aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to from 0.9 mmol / L to 1.7 mmol / L. In another aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to from 1.4 mmol / L to 2.4 mmol / L. In another aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to from 1.7 mmol / L to 2.5 mmol / L. In one aspect, the lactate level of a trauma patient in need of blood transfusion therapy is reduced to from 0.5 mmol / L to 1 mmol / L.

[0073] The methods of the present disclosure provide for and include reducing elevated lactate levels in hemorrhagic trauma patients in need of blood transfusion therapy to less than 4 mmol / L, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the lactate levels of trauma patients in need of blood transfusion therapy are reduced to less than 3 mmol / L. In yet another aspect, the lactate levels of trauma patients in need of blood transfusion therapy are reduced to less than 2.5 mmol / L. In another aspect, the lactate levels of the patient are reduced to less than 2.3 mmol / L. In another aspect, the lactate levels of trauma patients in need of blood transfusion therapy are reduced to less than 2 mmol / L. In another aspect, the lactate levels of trauma patients in need of blood transfusion therapy are reduced to less than 2 mmol / L. In another aspect, the lactate levels of trauma patients in need of blood transfusion therapy are reduced to less than 1.5 mmol / L. In another aspect, the lactate levels of trauma patients in need of blood transfusion therapy are reduced to less than 1 mmol / L.

[0074] It is also known that blood glucose levels are predictors of outcomes in several disease patterns and particularly in trauma patients. Compared to other critically ill patients, trauma patients with hyperglycemia are more likely to have adverse outcomes. See Kreutziger et al., “Admission blood glucose predicted hemorrhagic shock in multiple trauma patients,” Injury, 46:15 - 20 (2015) (incorporated herein by reference in its entirety). Studies evaluating the relationship of early hyperglycemia to trauma patients have examined early hyperglycemia at three possible cut-off values: glucose ≥ 110 mg / dL, glucose ≥ 150 mg / dL, and glucose ≥ 200 mg / dL. See Laird et al., “Relationship of early hyperglycemia to mortality in trauma patients,” J Trauma, 56:1058 - 62 (2004) (incorporated herein by reference in its entirety). Glucose levels of ≥ 200 mg / dL are significantly associated with higher infection and mortality in trauma patients, independent of injury characteristics. This is not the case at cut-off values of ≥ 110 mg / dL or ≥ 150 mg / dL. The reduced glucose levels in animals resuscitated with OR- and OCR-RBC compared to conventional RBC support the view that resuscitation with OR-RBC can significantly improve the clinical outcomes of patients. See Figure 8A and 8B 。

[0075] The methods of the present disclosure provide and include reducing glucose in trauma patients in need of blood transfusion therapy, including providing to the trauma patient oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, glucose is reduced by 10% to 90% compared to the infusion of blood stored under conventional conditions. In one aspect, the infusion of blood stored with OR reduces glucose by 10% to 50% compared to the infusion of blood stored under conventional conditions. In another aspect, the infusion of blood stored with OR reduces glucose by 20% to 40% compared to the infusion of blood stored under conventional conditions. In another aspect, the infusion of blood stored with OR reduces glucose by 50% to 90% compared to the infusion of blood stored under conventional conditions. In yet another aspect, the infusion of blood stored with OR reduces glucose by 60% to 90%. In another aspect, the infusion of blood stored with OR reduces glucose by 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, or 80% to 90% compared to the infusion of blood stored under conventional conditions. In another aspect, the infusion of blood stored with OR reduces glucose by at least 10% compared to the infusion of blood stored under conventional conditions. In another aspect, the infusion of blood stored with OR reduces glucose by at least 20%. In a further aspect, the infusion of blood stored with OR reduces glucose by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%.

[0076] The methods of the present disclosure provide and include reducing the glucose level of a trauma patient in need of blood transfusion therapy to about 70 mg / dL to about 120 mg / dL, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is about 70 mg / dL to about 110 mg / dL. In another aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is about 70 mg / dL to about 100 mg / dL. In another aspect, the glucose of the trauma patient after blood transfusion therapy with OR or OCR blood is about 90 mg / dL to about 120 mg / dL. In another aspect, the glucose of the trauma patient after blood transfusion therapy with OR or OCR blood is about 90 mg / dL to about 100 mg / dL.

[0077] The method of the present disclosure provides and includes reducing the glucose level of a trauma patient in need of blood transfusion therapy to 70 mg / dL to 120 mg / dL, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is 70 mg / dL to 110 mg / dL. In another aspect, the glucose of the patient is 70 mg / dL to 100 mg / dL. In another aspect, the glucose of the patient is 90 mg / dL to 120 mg / dL. In another aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is 90 mg / dL to 100 mg / dL.

[0078] The method of the present disclosure provides and includes reducing the glucose level of a trauma patient in need of blood transfusion therapy to less than 120 mg / dL, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In a further aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is less than 110 mmol / L, and in yet another aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is less than 100 mg / dL. In another aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is less than 200 mg / dL. In another aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is less than 90 mg / dL. In another aspect, the glucose of the patient after blood transfusion therapy with OR or OCR blood is less than 80 mg / dL.

[0079] In one aspect of the present disclosure, a patient is at an increased risk of complications caused by a blood transfusion therapy based on a pre-existing or potential medical condition. In one aspect, the patient has a pre-existing or potential medical condition selected from the group consisting of: diabetes, ischemic heart disease, systemic inflammatory syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoke inhalation, chronic obstructive pulmonary disease such as systemic inflammation caused by infection, coagulopathy, and autoimmune disease. In another aspect, the patient has one or more pre-existing or potential medical conditions selected from the group consisting of: diabetes, ischemic heart disease, systemic inflammatory syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoke inhalation, and chronic obstructive pulmonary disease such as systemic inflammation caused by infection, coagulopathy, and autoimmune disease. In another aspect, the patient has two or more pre-existing or potential medical conditions selected from the group consisting of: diabetes, ischemic heart disease, systemic inflammatory syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoke inhalation, chronic obstructive pulmonary disease such as systemic inflammation caused by infection, coagulopathy, and autoimmune disease. In another aspect, the patient has three or more pre-existing or potential medical conditions selected from the group consisting of: diabetes, ischemic heart disease, systemic inflammatory syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoke inhalation, chronic obstructive pulmonary disease such as systemic inflammation caused by infection, coagulopathy, and autoimmune disease.

[0080] During hemorrhagic shock, the patient experiences adverse events including liver injury or failure, kidney injury or failure, lung injury or failure, or a combination thereof. The present disclosure provides for and includes patients in need of blood transfusion therapy with OR or OCR RBCs who exhibit one or more adverse events selected from the group consisting of: liver injury or failure, kidney injury or failure, or lung injury or failure. The present disclosure provides for and includes patients in need who exhibit two or more adverse events selected from the group consisting of: liver injury or failure, kidney injury or failure, or lung injury or failure.

[0081] The methods of the present disclosure provide for and include reducing adverse events in trauma patients, including providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to trauma patients in need of blood transfusion therapy. In one aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 5%. In another aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 10%. In another aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 20%. In another aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 30%. In another aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 40%. In another aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 50%. In another aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 60%. In another aspect, adverse events are reduced by at least 70%. In another aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by at least 80%. In another aspect, adverse events are reduced by at least 90%. In a further aspect, adverse events after blood transfusion therapy with OR or OCR blood are reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95%. In one aspect, the adverse event after blood transfusion therapy with OR or OCR blood is liver injury or damage. In another aspect, the adverse event is lung injury or damage. In yet another aspect, the adverse event is kidney injury or damage. In a further aspect, the adverse event is liver injury, lung injury, kidney injury or a combination thereof.

[0082] Elevated levels of liver enzymes (including but not limited to aspartate aminotransferase (AST) and alanine aminotransferase (ALT)) indicate some form of liver damage, shock or injury. The methods of the present disclosure provide for and include reducing elevated levels of liver enzymes in trauma patients, including providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to trauma patients.

[0083] The method of the present disclosure provides and includes reducing the AST level in trauma patients in need of blood transfusion treatment, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the AST level is reduced by at least 5% relative to the AST level of a patient infused with conventionally stored blood. In another aspect, the AST level is reduced by at least 10% relative to the AST level of a patient infused with conventionally stored blood. In another aspect, the AST level is reduced by at least 20% relative to the AST level of a patient infused with conventionally stored blood. In another aspect, the AST level is reduced by at least 30% relative to the AST level of a patient infused with conventionally stored blood. In another aspect, the AST level is reduced by at least 40%. In another aspect, the AST level is reduced by at least 50% relative to the AST level of a patient infused with conventionally stored blood. In another aspect, the AST level is reduced by at least 60%. In another aspect, the AST level is reduced by at least 70% relative to the AST level of a patient infused with conventionally stored blood. In yet another aspect, the AST level is reduced by at least 80%. In a further aspect, the AST level is reduced by at least 90% relative to the AST level of a patient infused with conventionally stored blood. In a further aspect, the AST level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the AST level of a patient infused with conventionally stored blood.

[0084] The method of the present disclosure provides and includes reducing the AST level in trauma patients in need of blood transfusion treatment by 1.5 to 10 times, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the AST level is reduced by 2 to 3 times relative to the AST level of a patient infused with conventionally stored blood. In another aspect, the AST level is reduced by 3 to 4 times. In another aspect, the AST level is reduced by 4 to 10 times. In another aspect, the AST level is reduced by 6 to 9 times relative to the AST level of a patient infused with conventionally stored blood. In a further aspect, the AST level is reduced by 2 to 5 times. In another aspect, the AST level is reduced by 10 to 50 times relative to the AST level of a patient infused with conventionally stored blood.

[0085] The method of the present disclosure provides and includes reducing the AST level in a trauma patient in need of blood transfusion therapy by at least 1.5-fold, including providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the AST level is reduced by at least 2-fold relative to the AST level of a patient receiving conventionally stored blood. In another aspect, the AST level is reduced by at least 3-fold relative to the AST level of a patient receiving conventionally stored blood. In another aspect, the AST level is reduced by at least 4-fold relative to the AST level of a patient receiving conventionally stored blood. In another aspect, the AST level is reduced by at least 5-fold relative to the AST level of a patient receiving conventionally stored blood. In a further aspect, the AST level is reduced by at least 6-fold. In another aspect, the AST level is reduced by at least 7-fold relative to the AST level of a patient receiving conventionally stored blood. In another aspect, the AST level is reduced by at least 8-fold. In another aspect, the AST level is reduced by at least 9-fold relative to the AST level of a patient receiving conventionally stored blood. In another aspect, the AST level is reduced by at least 10-fold relative to the AST level of a patient receiving conventionally stored blood. In a further aspect, the AST level is reduced by at least 50-fold relative to the AST level of a patient receiving conventionally stored blood.

[0086] The method of the present disclosure provides and includes reducing ALT levels in trauma patients in need of blood transfusion treatment, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the ALT level is reduced by at least 5% relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 10% relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 20% relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 30% relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 40% relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 50% relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 60% relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 70% relative to the ALT level of a patient receiving conventionally stored blood. In yet another aspect, the ALT level is reduced by at least 80% relative to the ALT level of a patient receiving conventionally stored blood. In a further aspect, the ALT level is reduced by at least 90%. In a further aspect, the ALT level is reduced by at least 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the ALT level of a patient receiving conventionally stored blood.

[0087] The method of the present disclosure provides and includes reducing the ALT level in a trauma patient in need of blood transfusion treatment by 1.5 to 10 times, including providing to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the ALT level is reduced by 2 to 3 times relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by 3 to 4 times relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by 4 to 10 times relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by 6 to 9 times relative to the ALT level of a patient receiving conventionally stored blood. In a further aspect, the ALT level is reduced by 2 to 5 times. In another aspect, the ALT level is reduced by 10 to 50 times relative to the ALT level of a patient receiving conventionally stored blood.

[0088] The method of the present disclosure provides and includes reducing the ALT level in a trauma patient in need of blood transfusion therapy by at least 1.5-fold, including providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the ALT level is reduced by at least 2-fold relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 3-fold relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 4-fold relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 5-fold relative to the ALT level of a patient receiving conventionally stored blood. In a further aspect, the ALT level is reduced by at least 6-fold relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 7-fold relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 8-fold relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 9-fold relative to the ALT level of a patient receiving conventionally stored blood. In another aspect, the ALT level is reduced by at least 10-fold relative to the ALT level of a patient receiving conventionally stored blood. In a further aspect, the ALT level is reduced by at least 50-fold relative to the ALT level of a patient receiving conventionally stored blood.

[0089] Markers of kidney function during and after hemorrhagic trauma include urinary neutrophil gelatinase-associated lipocalin (u-NGAL), serum creatinine, and blood urea nitrogen (BUN). See Treeprasertsuk et al., “Urine neutrophil gelatinase-associated lipocalin: a diagnostic and prognostic marker for acute kidney injury (AKI) in hospitalized cirrhotic patients with AKI-prone conditions,” BMC Gastroenterol 15:140 (2015) (incorporated herein by reference in its entirety). Gene expression analyses reported in over 150 different studies conducted in AKI models from multiple species ranging from rodents to humans consistently reveal that the NGAL gene is one of the most significantly upregulated genes in the kidney shortly after ischemic or nephrotoxic injury. See Ciccia et al., “Pediatric acute kidney injury: prevalence, impact and management challenges,” Int J Nephrol Renovasc Dis, 10:77-84 (2017) (incorporated herein by reference in its entirety). Similarly, serum creatinine levels can vary according to age, race, and body size, however, an elevated creatinine level indicates kidney damage. A creatinine level above 1.2 in females and above 1.4 in males may be an early sign of kidney damage. It can be seen that an elevated blood urea nitrogen (BUN) is associated with kidney disease or failure as well as congestive heart failure, shock, and gastrointestinal bleeding. If the BUN level is above 100 mg / dL, it indicates severe kidney damage. A decrease in the BUN level is also a concern and may indicate fluid overload, trauma, surgery, opioids, malnutrition, and anabolic steroid use.See Pagana, “Mosby's Manual of Diagnostic and Laboratory Tests,” St. Louis Mosby, Inc., (1998); and Gowda, et al., “Markers of renal function tests,” N Am J Med Sci. 2(4):170–173 (2010) (incorporated herein by reference in its entirety). As provided in the present disclosure, compared with conventional RBCs, u-NGAL in animals resuscitated with OR- and OCR-RBCs ( Figure 16A and 16B ), serum creatinine ( Figure 11A and 11B ), and BUN ( Figure 12A and 12B ) levels are reduced, indicating that resuscitation with OR-RBCs can significantly improve the clinical outcomes of patients.

[0090] The method of the present disclosure provides and includes reducing the level of urinary neutrophil gelatinase-associated lipocalin (u-NGAL) in patients in need of blood transfusion treatment, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the u-NGAL level is reduced by at least 5% relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 10% relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 20% relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 30%. In another aspect, the u-NGAL level is reduced by at least 40% relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 50% relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 60% relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 70% relative to the u-NGAL level of a patient who receives conventionally stored blood. In yet another aspect, the u-NGAL level is reduced by at least 80% relative to the u-NGAL level of a patient who receives conventionally stored blood. In a further aspect, the u-NGAL level is reduced by at least 90% relative to the u-NGAL level of a patient who receives conventionally stored blood. In a further aspect, the u-NGAL level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the u-NGAL level of a patient who receives conventionally stored blood.

[0091] The method of the present disclosure provides and includes reducing the urinary neutrophil gelatinase-associated lipocalin (u-NGAL) level in a patient in need of blood transfusion therapy by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the u-NGAL level is reduced by 2 to 3-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by 3 to 4-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by 4 to 10-fold. In another aspect, the u-NGAL level is reduced by 6 to 9-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In a further aspect, the u-NGAL level is reduced by 2 to 5-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by 10 to 50-fold relative to the u-NGAL level of a patient who receives conventionally stored blood.

[0092] The method of the present disclosure provides and includes reducing the urinary neutrophil gelatinase-associated lipocalin (u-NGAL) level in a patient in need of blood transfusion therapy by at least 1.5-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the u-NGAL level is reduced by at least 2-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 3-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 4-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 5-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In a further aspect, the u-NGAL level is reduced by at least 6-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 7-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 8-fold relative to the u-NGAL level of a patient who receives conventionally stored blood. In another aspect, the u-NGAL level is reduced by at least 9-fold. In another aspect, the u-NGAL level is reduced by at least 10-fold. In a further aspect, the u-NGAL level is reduced by at least 50-fold.

[0093] The method of the present disclosure provides and includes reducing the serum creatinine level in a patient in need of blood transfusion therapy, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the serum creatinine level is reduced by at least 5% relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 10% relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 20% relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 30% relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 40% relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 50% relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 60% relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 70% relative to the serum creatinine level of a patient receiving conventionally stored blood. In yet another aspect, the serum creatinine level is reduced by at least 80% relative to the serum creatinine level of a patient receiving conventionally stored blood. In a further aspect, the serum creatinine level is reduced by at least 90% relative to the serum creatinine level of a patient receiving conventionally stored blood. In a further aspect, the serum creatinine level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the serum creatinine level of a patient receiving conventionally stored blood.

[0094] The method of the present disclosure provides and includes reducing the serum creatinine level in a patient in need of blood transfusion therapy by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the serum creatinine level is reduced by 2 to 3-fold relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by 3 to 4-fold relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by 4 to 10-fold relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by 6 to 9-fold relative to the serum creatinine level of a patient receiving conventionally stored blood. In a further aspect, the serum creatinine level is reduced by 2 to 5-fold relative to the serum creatinine level of a patient receiving conventionally stored blood. In another aspect, the serum creatinine level is reduced by 10 to 50-fold relative to the serum creatinine level of a patient receiving conventionally stored blood.

[0095] The method of the present disclosure provides and includes reducing the serum creatinine level of a patient in need of blood transfusion therapy by at least 1.5-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the serum creatinine level is reduced by at least 2-fold relative to the serum creatinine level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 3-fold relative to the AST level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 4-fold relative to the serum creatinine level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 5-fold. In a further aspect, the serum creatinine level is reduced by at least 6-fold relative to the serum creatinine level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 7-fold relative to the serum creatinine level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 8-fold relative to the serum creatinine level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 9-fold relative to the serum creatinine level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 10-fold relative to the serum creatinine level of a patient who receives conventionally stored blood.

[0096] The method of the present disclosure provides and includes reducing the serum creatinine level of a patient in need of blood transfusion therapy to 0.5 to 1.5 mg / dL, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the serum creatinine level is reduced to 0.5 mg / dL to 1 mg / dL relative to the serum creatinine level of a patient who receives conventionally stored blood. In one aspect, the serum creatinine level is reduced to 0.8 mg / dL to 1 mg / dL relative to the serum creatinine level of a patient who receives conventionally stored blood. In another aspect, the serum creatinine level is reduced to 0.7 mg / dL to 1.5 mg / dL relative to the serum creatinine level of a patient who receives conventionally stored blood.

[0097] The method of the present disclosure provides and includes reducing the serum creatinine level of a patient in need of blood transfusion therapy to less than 1.5 mg / dL, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the serum creatinine level is reduced to less than 1.4 mg / dL relative to the serum creatinine level of a patient who receives conventionally stored blood. In one aspect, the serum creatinine level is reduced to less than 1 mg / dL. In another aspect, the serum creatinine level is reduced to less than 0.8 mg / dL relative to the serum creatinine level of a patient who receives conventionally stored blood.

[0098] The methods of the present disclosure provide and include reducing BUN levels in a patient, including providing to a patient in need of a blood transfusion oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the BUN level is reduced by at least 5% relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 10% relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 20% relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 30% relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 40% relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 50% relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 60% relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 70% relative to the BUN level of a patient receiving conventionally stored blood. In yet another aspect, the BUN level is reduced by at least 80% relative to the BUN level of a patient receiving conventionally stored blood. In a further aspect, the BUN level is reduced by at least 90% relative to the BUN level of a patient receiving conventionally stored blood. In a further aspect, for the BUN level of a patient receiving conventionally stored blood, the BUN level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95%.

[0099] The methods of the present disclosure provide and include reducing the BUN level in a patient by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the BUN level is reduced by 2 to 3-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by 3 to 4-fold. In another aspect, the BUN level is reduced by 4 to 10-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by 6 to 9-fold relative to the BUN level of a patient receiving conventionally stored blood. In a further aspect, the BUN level is reduced by 2 to 5-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by 10 to 100-fold relative to the BUN level of a patient receiving conventionally stored blood.

[0100] The method of the present disclosure provides and includes reducing the BUN level in a patient by at least 1.5-fold, including providing to a patient in need of blood transfusion treatment oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the BUN level is reduced by at least 2-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 3-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 4-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 5-fold relative to the BUN level of a patient receiving conventionally stored blood. In a further aspect, the BUN level is reduced by at least 6-fold. In another aspect, the BUN level is reduced by at least 7-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 8-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 9-fold relative to the BUN level of a patient receiving conventionally stored blood. In another aspect, the BUN level is reduced by at least 10-fold relative to the BUN level of a patient receiving conventionally stored blood.

[0101] The methods of the present disclosure provide and include reducing the percentage of CD45+ neutrophils in a patient, including providing to a patient in need of a blood transfusion oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the percentage of CD45+ neutrophils is reduced by at least 5% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 10% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 20% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 30% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 40% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 50% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 60% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 70% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In yet another aspect, the percentage of CD45+ neutrophils is reduced by at least 80% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In a further aspect, the percentage of CD45+ neutrophils is reduced by at least 90% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion. In a further aspect, the percentage of CD45+ neutrophils is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the CD45+ neutrophil level in a patient who receives a conventionally stored blood transfusion.

[0102] The method of the present disclosure provides for and includes reducing the percentage of CD45+ neutrophils in a patient in need of blood transfusion therapy by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the percentage of CD45+ neutrophils is reduced by 2 to 3-fold relative to the CD45+ neutrophil level in a patient receiving conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by 3 to 4-fold relative to the CD45+ neutrophil level in a patient receiving conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by 4 to 10-fold relative to the CD45+ neutrophil level in a patient receiving conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by 6 to 9-fold relative to the CD45+ neutrophil level in a patient receiving conventionally stored blood. In a further aspect, the percentage of CD45+ neutrophils is reduced by 2 to 5-fold relative to the CD45+ neutrophil level in a patient receiving conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by 10 to 50-fold relative to the CD45+ neutrophil level in a patient receiving conventionally stored blood.

[0103] The method of the present disclosure provides for and includes reducing the percentage of CD45+ neutrophils in a trauma patient in need of blood transfusion therapy by at least 1.5-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the percentage of CD45+ neutrophils is reduced by at least 2-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 3-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 4-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 5-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In a further aspect, the percentage of CD45+ neutrophils is reduced by at least 6-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 7-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 8-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 9-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 10-fold relative to the CD45+ neutrophil level in a patient infused with conventionally stored blood.

[0104] The method of the present disclosure provides and includes reducing CXCL1 levels in a patient in need of blood transfusion treatment, including providing to the patient in need of blood transfusion treatment oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CXCL1 level is reduced by at least 5% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 10% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 20% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 30% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 40% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 50% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 60% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 70% relative to the CXCL1 level of a patient receiving conventionally stored blood. In yet another aspect, the CXCL1 level is reduced by at least 80% relative to the CXCL1 level of a patient receiving conventionally stored blood. In a further aspect, the CXCL1 level is reduced by at least 90% relative to the CXCL1 level of a patient receiving conventionally stored blood. In another aspect, the CXCL1 level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the CXCL1 level of a patient receiving conventionally stored blood.

[0105] The methods of the present disclosure provide for and include reducing the CXCL1 level in a patient by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CXCL1 level is reduced by 2 to 3-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by 3 to 4-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by 4 to 10-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by 6 to 9-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In a further aspect, the CXCL1 level is reduced by 2 to 5-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by 10 to 100-fold relative to the CXCL1 level of a patient who receives conventionally stored blood.

[0106] The methods of the present disclosure provide for and include reducing the CXCL1 level in a patient by at least 1.5-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CXCL1 level is reduced by at least 2-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 3-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 4-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 5-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In a further aspect, the CXCL1 level is reduced by at least 6-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 7-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 8-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 9-fold relative to the CXCL1 level of a patient who receives conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 10-fold relative to the CXCL1 level of a patient who receives conventionally stored blood.

[0107] Although limited by the sensitivity of the test, the IL-6 level in healthy patients is typically less than 5x10 -12grams per liter (picograms per liter or pg / L). In the present disclosure, the IL-6 level of a patient in need is at least 5 pg / L. In another aspect, the IL-6 level of a patient is at least 10 pg / L, 20 pg / L, 40 pg / L, 60 pg / L, 80 pg / L, 100 pg / L, or 150 pg / L. In yet another aspect, the IL-6 level of a patient in need is from 10 pg / L to 50 pg / L, 50 pg / L to 100 mg / L, 100 pg / L to 200 mg / L, or 10 pg / L to 200 mg / L. The methods of the present disclosure provide and include reducing the IL-6 level in a patient in need of blood transfusion therapy, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-6 level is reduced by at least 5% relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 10% relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 20%. In another aspect, the IL-6 level is reduced by at least 30% relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 40% relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 50% relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 60% relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 70% relative to the IL-6 level of a patient receiving conventionally stored blood. In yet another aspect, the IL-6 level is reduced by at least 80% relative to the IL-6 level of a patient receiving conventionally stored blood. In a further aspect, the IL-6 level is reduced by at least 90% relative to the IL-6 level of a patient receiving conventionally stored blood. In a further aspect, the IL-6 level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90%, or 90% to 95% relative to the IL-6 level of a patient receiving conventionally stored blood.

[0108] The method of the present disclosure provides and includes reducing the IL-6 level in a patient by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-6 level is reduced by 2 to 3-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by 3 to 4-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by 4 to 10-fold. In another aspect, the IL-6 level is reduced by 6 to 9-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In a further aspect, the IL-6 level is reduced by 2 to 5-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by 10 to 100-fold relative to the IL-6 level of a patient receiving conventionally stored blood.

[0109] The method of the present disclosure provides and includes reducing the IL-6 level in a patient by at least 1.5-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-6 level is reduced by at least 2-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 3-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 4-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 5-fold. In a further aspect, the IL-6 level is reduced by at least 6-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 7-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 8-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 9-fold relative to the IL-6 level of a patient receiving conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 10-fold relative to the IL-6 level of a patient receiving conventionally stored blood.

[0110] In one aspect of the present disclosure, the patient in need is a patient suffering from sepsis.

[0111] In one aspect of the present disclosure, the patient in need is a patient suffering from inflammation. In another aspect, the patient has systemic inflammation. In another aspect, the patient has chronic inflammation. In another aspect, the patient has acute inflammation.

[0112] In one aspect of the present disclosure, a patient with inflammation is a patient with sickle cell disease. In a further aspect, a patient with sickle cell disease has sickle cell anemia. In another aspect, a patient with sickle cell disease has a sickle cell crisis. In yet another aspect, the patient has a type of sickle cell disease selected from the group consisting of hemoglobin SS, hemoglobin SC, hemoglobin SB+β-thalassemia, hemoglobin SB(β-0) thalassemia, hemoglobin SD, hemoglobin SE, and hemoglobin SO.

[0113] The present disclosure provides a method for preventing or reducing the number of vaso-occlusive episodes in a sickle cell patient in need thereof, comprising providing a previously stored oxygen-reduced blood to a patient with sickle cell disease, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage. In one aspect, the vaso-occlusive episodes are reduced by at least 10% compared to sickle cell patients receiving conventionally stored blood. In another aspect, the number of vaso-occlusive episodes is reduced by at least 10% over a period of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, or 50 weeks.

[0114] The present disclosure provides a reduction in the adhesion of red blood cells to endothelial cells expressing thrombospondin in a sickle cell patient in need thereof. The present disclosure also provides a reduction in the adhesion of red blood cells to endothelial cells expressing vascular cell adhesion molecule-1 (VCAM-1) in a patient in need thereof. Further, the present disclosure provides a reduction in the adhesion of red blood cells to endothelial cells expressing laminin in a patient in need thereof. In one aspect, the adhesion is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% compared to the adhesion of conventionally stored blood. In another aspect, the adhesion is reduced by 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, or 80% to 90% compared to the adhesion of conventionally stored blood.

[0115] The present disclosure provides methods for improving transfusion in sickle cell patients in need, including providing a patient with sickle cell disease with previously stored oxygen-reduced blood, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage. In one aspect, the patient has dactylitis (hand-foot syndrome). In another aspect, the methods of the present disclosure provide for reducing the occurrence of dactylitis. In another aspect, compared to patients receiving transfusions of conventionally stored RBCs, the occurrence of dactylitis is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80%. In another aspect, the present disclosure provides for reducing the number of pain crises in a patient compared to a patient treated with conventionally stored RBCs. In yet another aspect, the present disclosure provides for reducing complications of anemia, the complications selected from the group consisting of fatigue, irritability, dizziness, dyspnea, pallor of skin color, jaundice, slow growth, and developmental delay. In another aspect, the present disclosure provides for reducing infection. In yet another aspect, the present disclosure provides for preventing infection. In a further aspect, the present disclosure provides for reducing spleen damage. In one aspect, the spleen damage is splenic sequestration. In another aspect, the spleen damage is splenomegaly. In a further aspect, the present disclosure provides for reducing or preventing stroke.

[0116] In one aspect of the present disclosure, the patient in need is a febrile patient. In another aspect, the patient in need has a fever of at least 38 °C. In another aspect, the patient in need has a fever of about 39 °C, 40 °C, 41 °C, or 42 °C. In another aspect, the patient has a fever of 38 °C to 42 °C. In another aspect, compared to a patient without a fever, the patient in need has a fever and an increased level of C-reactive protein (CRP). In another aspect, compared to a patient without a fever, the level of one or more inflammatory mediators selected from the group consisting of IL-6, IL-8, IL-10, and granulocyte colony-stimulating factor (G-CSF) is increased in the febrile patient. In another aspect, compared to a patient without a fever, the level of one or more inflammatory mediators selected from the group consisting of CCL-5 and CXCL-10 is decreased in the febrile patient.

[0117] In one aspect of the present disclosure, the patient in need is a patient at risk of fever. In another aspect, the patient has a fever of 38 °C to 42 °C. In another aspect, compared to a healthy person, the patient in need at risk of fever has an increased level of C-reactive protein (CRP). In another aspect, compared to a healthy person, the level of one or more inflammatory mediators selected from the group consisting of IL-6, IL-8, IL-10, and granulocyte colony-stimulating factor (G-CSF) is increased in the patient in need. In another aspect, compared to a healthy patient, the level of one or more inflammatory mediators selected from the group consisting of CCL-5 and CXCL-10 is decreased in the patient in need.

[0118] Although limited by the test sensitivity, the CRP level in healthy patients is usually less than 3 mg / L. In one aspect, the CRP level in patients in need is at least 3 mg / L. In another aspect, the CRP level in patients is at least 4 mg / L, 5 mg / L, 6 mg / L, 7 mg / L, 8 mg / L, 9 mg / L or 10 mg / L. In yet another aspect, the CRP level in patients in need is from 3 mg / L to 5 mg / L, from 3 mg / L to 10 mg / L, from 5 mg / L to 10 mg / L or from 3 mg / L to 8 mg / L.

[0119] The methods of the present disclosure provide and include reducing the CRP level in patients in need of blood transfusion therapy, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CRP level is restored to the normal level of a healthy person. In one aspect, the CRP level is reduced by at least 5% relative to the CRP level of a patient who receives conventionally stored blood. In another aspect, the CRP level is reduced by at least 10% relative to the CRP level of a patient who receives conventionally stored blood. In another aspect, the CRP level is reduced by at least 20%. In another aspect, the CRP level is reduced by at least 30% relative to the CRP level of a patient who receives conventionally stored blood. In another aspect, the CRP level is reduced by at least 40% relative to the CRP level of a patient who receives conventionally stored blood. In another aspect, the CRP level is reduced by at least 50% relative to the CRP level of a patient who receives conventionally stored blood. In another aspect, the CRP level is reduced by at least 60% relative to the CRP level of a patient who receives conventionally stored blood. In another aspect, the CRP level is reduced by at least 70% relative to the CRP level of a patient who receives conventionally stored blood. In yet another aspect, the CRP level is reduced by at least 80% relative to the CRP level of a patient who receives conventionally stored blood. In a further aspect, the CRP level is reduced by at least 90% relative to the CRP level of a patient who receives conventionally stored blood. In a further aspect, the CRP level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the CRP level of a patient who receives conventionally stored blood.

[0120] The method of the present disclosure provides and includes reducing the CRP level in a patient in need by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CRP level is reduced by 2 to 3-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by 3 to 4-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by 4 to 10-fold. In another aspect, the CRP level is reduced by 6 to 9-fold relative to the CRP level of a patient receiving conventionally stored blood. In a further aspect, the CRP level is reduced by 2 to 5-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by 10 to 100-fold relative to the CRP level of a patient receiving conventionally stored blood.

[0121] The method of the present disclosure provides and includes reducing the CRP level in a patient in need by at least 1.5-fold, including providing to a patient in need of blood transfusion therapy oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CRP level is reduced by at least 2-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by at least 3-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by at least 4-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by at least 5-fold. In a further aspect, the CRP level is reduced by at least 6-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by at least 7-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by at least 8-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by at least 9-fold relative to the CRP level of a patient receiving conventionally stored blood. In another aspect, the CRP level is reduced by at least 10-fold relative to the CRP level of a patient receiving conventionally stored blood.

[0122] Although limited by the test sensitivity, the IL-8 level in healthy patients is typically less than 40 picograms per liter (pg / L). In one aspect, the IL-8 level in a patient in need is at least 40 pg / L. In another aspect, the IL-8 level of the patient is at least 50 pg / L, 5 pg / L, 6 pg / L, 7 pg / L, 8 pg / L, 9 pg / L, or 10 pg / L. In yet another aspect, the CRP level in a patient in need is 30 pg / L to 50 pg / L, 40 pg / L to 100 pg / L, 50 pg / L to 100 pg / L, or 60 pg / L to 200 pg / L.

[0123] The method of the present disclosure provides and includes reducing the IL-8 level in a patient in need of blood transfusion treatment, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-8 level is restored to the normal level of a healthy person. In one aspect, the IL-8 level is reduced by at least 5% relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 10% relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 20%. In another aspect, the IL-8 level is reduced by at least 30% relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 40% relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 50% relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 60% relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 70% relative to the IL-8 level of a patient infused with conventionally stored blood. In yet another aspect, the IL-8 level is reduced by at least 80% relative to the IL-8 level of a patient infused with conventionally stored blood. In a further aspect, the IL-8 level is reduced by at least 90% relative to the IL-8 level of a patient infused with conventionally stored blood. In a further aspect, the IL-8 level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the IL-8 level of a patient infused with conventionally stored blood.

[0124] The method of the present disclosure provides and includes reducing the IL-8 level in a patient in need by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-8 level is reduced by 2 to 3-fold relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by 3 to 4-fold relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by 4 to 10-fold. In another aspect, the IL-8 level is reduced by 6 to 9-fold relative to the IL-8 level of a patient infused with conventionally stored blood. In a further aspect, the IL-8 level is reduced by 2 to 5-fold relative to the IL-8 level of a patient infused with conventionally stored blood. In another aspect, the IL-8 level is reduced by 10 to 100-fold relative to the IL-8 level of a patient infused with conventionally stored blood.

[0125] The method of the present disclosure provides and includes reducing the IL-8 level in a patient in need by at least 1.5-fold, including providing to a patient in need of blood transfusion therapy oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-8 level is reduced by at least 2-fold relative to the IL-8 level of a patient receiving conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 3-fold relative to the IL-8 level of a patient receiving conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 4-fold relative to the IL-8 level of a patient receiving conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 5-fold. In a further aspect, the IL-8 level is reduced by at least 6-fold relative to the IL-8 level of a patient receiving conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 7-fold relative to the IL-8 level of a patient receiving conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 8-fold relative to the IL-8 level of a patient receiving conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 9-fold relative to the IL-8 level of a patient receiving conventionally stored blood. In another aspect, the IL-8 level is reduced by at least 10-fold relative to the IL-8 level of a patient receiving conventionally stored blood.

[0126] The IL-10 level in healthy individuals is less than about 20 pg / L. See Kleiner, G., et al., "Cytokine Levels in the Serum of Healthy Subjects," Mediators of Inflammation, 1-6 (2013). In the present disclosure, the IL-10 level in a patient in need is at least 15 pg / L. In another aspect, the IL-10 level of the patient is at least 20 pg / L, 40 pg / L, 50 pg / L, 60 pg / L, 80 pg / L, 100 pg / L, or 150 pg / L. In yet another aspect, the IL-10 level in a patient in need is from 20 pg / L to 50 pg / L, 50 mg / L to 100 mg / L, 100 mg / L to 200 mg / L, or 15 mg / L to 200 mg / L.

[0127] The method of the present disclosure provides and includes reducing IL-10 levels in a patient in need of blood transfusion therapy, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-10 levels are restored to normal levels of a healthy person. In one aspect, the IL-10 levels are reduced by at least 5% relative to the IL-10 levels of patients receiving conventionally stored blood. In another aspect, the IL-10 levels are reduced by at least 10% relative to the IL-10 levels of patients receiving conventionally stored blood. In another aspect, the IL-10 levels are reduced by at least 20%. In another aspect, the IL-10 levels are reduced by at least 30% relative to the IL-10 levels of patients receiving conventionally stored blood. In another aspect, the IL-10 levels are reduced by at least 40% relative to the IL-10 levels of patients receiving conventionally stored blood. In another aspect, the IL-10 levels are reduced by at least 50% relative to the IL-10 levels of patients receiving conventionally stored blood. In another aspect, the IL-10 levels are reduced by at least 60% relative to the IL-10 levels of patients receiving conventionally stored blood. In another aspect, the IL-10 levels are reduced by at least 70% relative to the IL-10 levels of patients receiving conventionally stored blood. In yet another aspect, the IL-10 levels are reduced by at least 80% relative to the IL-10 levels of patients receiving conventionally stored blood. In a further aspect, the IL-10 levels are reduced by at least 90% relative to the IL-10 levels of patients receiving conventionally stored blood. In a further aspect, the IL-10 levels are reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the IL-10 levels of patients receiving conventionally stored blood.

[0128] The methods of the present disclosure provide and include reducing the IL-10 level in a patient in need by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-10 level is reduced by 2 to 3-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by 3 to 4-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by 4 to 10-fold. In another aspect, the IL-10 level is reduced by 6 to 9-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In a further aspect, the IL-10 level is reduced by 2 to 5-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by 10 to 100-fold relative to the IL-10 level of a patient infused with conventionally stored blood.

[0129] The methods of the present disclosure provide and include reducing the IL-10 level in a patient in need by at least 1.5-fold, including providing to a patient in need of blood transfusion therapy oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-10 level is reduced by at least 2-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by at least 3-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by at least 4-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by at least 5-fold. In a further aspect, the IL-10 level is reduced by at least 6-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by at least 7-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by at least 8-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by at least 9-fold relative to the IL-10 level of a patient infused with conventionally stored blood. In another aspect, the IL-10 level is reduced by at least 10-fold relative to the IL-10 level of a patient infused with conventionally stored blood.

[0130] The method of the present disclosure provides and includes reducing G-CSF levels in a patient in need of blood transfusion therapy, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the G-CSF level is restored to the normal level of a healthy person. In one aspect, the G-CSF level is reduced by at least 5% relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 10% relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 20%. In another aspect, the G-CSF level is reduced by at least 30% relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 40% relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 50% relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 60% relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 70% relative to the G-CSF level of a patient infused with conventionally stored blood. In yet another aspect, the G-CSF level is reduced by at least 80% relative to the G-CSF level of a patient infused with conventionally stored blood. In a further aspect, the G-CSF level is reduced by at least 90% relative to the G-CSF level of a patient infused with conventionally stored blood. In a further aspect, the G-CSF level is reduced by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the G-CSF level of a patient infused with conventionally stored blood.

[0131] The method of the present disclosure provides and includes reducing the G-CSF level in a patient by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the G-CSF level is reduced by 2 to 3-fold relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by 3 to 4-fold relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by 4 to 10-fold. In another aspect, the G-CSF level is reduced by 6 to 9-fold relative to the G-CSF level of a patient infused with conventionally stored blood. In a further aspect, the G-CSF level is reduced by 2 to 5-fold relative to the G-CSF level of a patient infused with conventionally stored blood. In another aspect, the G-CSF level is reduced by 10 to 100-fold relative to the G-CSF level of a patient infused with conventionally stored blood.

[0132] The method of the present disclosure provides for and includes reducing the G-CSF level in a patient in need by at least 1.5-fold, including providing to a patient in need of blood transfusion treatment oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the G-CSF level is reduced by at least 2-fold relative to the G-CSF level of a patient receiving conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 3-fold relative to the G-CSF level of a patient receiving conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 4-fold relative to the G-CSF level of a patient receiving conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 5-fold. In a further aspect, the G-CSF level is reduced by at least 6-fold relative to the G-CSF level of a patient receiving conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 7-fold relative to the G-CSF level of a patient receiving conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 8-fold relative to the G-CSF level of a patient receiving conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 9-fold relative to the G-CSF level of a patient receiving conventionally stored blood. In another aspect, the G-CSF level is reduced by at least 10-fold relative to the G-CSF level of a patient receiving conventionally stored blood.

[0133] The method of the present disclosure provides and includes increasing CCL-5 levels in a patient in need of blood transfusion therapy, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CCL-5 level is restored to the normal level of a healthy person. In one aspect, the CCL-5 level is increased by at least 5% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 10% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 20%. In another aspect, the CCL-5 level is increased by at least 30% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 40% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 50% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 60% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 70% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In yet another aspect, the CCL-5 level is increased by at least 80% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In a further aspect, the CCL-5 level is increased by at least 90% relative to the CCL-5 level of a patient transfused with conventionally stored blood. In a further aspect, the CCL-5 level is increased by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the CCL-5 level of a patient transfused with conventionally stored blood.

[0134] The method of the present disclosure provides and includes increasing the CCL-5 level in a patient in need by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CCL-5 level is reduced by 2 to 3-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by 3 to 4-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by 4 to 10-fold. In another aspect, the CCL-5 level is increased by 6 to 9-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In a further aspect, the CCL-5 level is increased by 2 to 5-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by 10 to 100-fold relative to the CCL-5 level of a patient infused with conventionally stored blood.

[0135] The method of the present disclosure provides and includes increasing the CCL-5 level in a patient in need by at least 1.5-fold, including providing to a patient in need of blood transfusion therapy oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CCL-5 level is increased by at least 2-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 3-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 4-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 5-fold. In a further aspect, the CCL-5 level is increased by at least 6-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 7-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 8-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 9-fold relative to the CCL-5 level of a patient infused with conventionally stored blood. In another aspect, the CCL-5 level is increased by at least 10-fold relative to the CCL-5 level of a patient infused with conventionally stored blood.

[0136] The method of the present disclosure provides and includes increasing CXCL-10 levels in a patient in need of blood transfusion therapy, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CXCL-10 levels are restored to normal levels of a healthy person. In one aspect, the CXCL-10 levels are increased by at least 5% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In another aspect, the CXCL-10 levels are increased by at least 10% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In another aspect, the CXCL-10 levels are increased by at least 20%. In another aspect, the CXCL-10 levels are increased by at least 30% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In another aspect, the CXCL-10 levels are increased by at least 40% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In another aspect, the CXCL-10 levels are increased by at least 50% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In another aspect, the CXCL-10 levels are increased by at least 60% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In another aspect, the CXCL-10 levels are increased by at least 70% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In yet another aspect, the CXCL-10 levels are increased by at least 80% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In a further aspect, the CXCL-10 levels are increased by at least 90% relative to the CXCL-10 levels of patients receiving conventionally stored blood. In a further aspect, the CXCL-10 levels are increased by 1% to 10%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90% or 90% to 95% relative to the CXCL-10 levels of patients receiving conventionally stored blood.

[0137] The method of the present disclosure provides and includes increasing the CXCL-10 level in a patient in need by 1.5 to 10-fold, including providing to the patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CXCL-10 level is reduced by 2 to 3-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by 3 to 4-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by 4 to 10-fold. In another aspect, the CXCL-10 level is increased by 6 to 9-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In a further aspect, the CXCL-10 level is increased by 2 to 5-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by 10 to 100-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood.

[0138] The method of the present disclosure provides and includes increasing the CXCL-10 level in a patient in need by at least 1.5-fold, including providing to a patient in need of blood transfusion therapy oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CXCL-10 level is increased by at least 2-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by at least 3-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by at least 4-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by at least 5-fold. In a further aspect, the CXCL-10 level is increased by at least 6-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by at least 7-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by at least 8-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by at least 9-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood. In another aspect, the CXCL-10 level is increased by at least 10-fold relative to the CXCL-10 level of a patient infused with conventionally stored blood.

[0139] As used herein, the terms "higher", "greater", or "increased" mean that the measured value of oxygen-reduced and anoxic stored blood is at least 1 standard deviation greater than the measured value of conventionally stored blood that is otherwise equivalently processed, with a sample size of at least 2 for each comparison measurement condition.

[0140] As used herein, the terms "reduced", "reduction", "lower", "decrease", or "less" mean that the measured value of oxygen-reduced and anoxic-stored blood is at least 1 standard deviation lower than the measured value of normoxic or hyperoxic conventionally stored blood that is otherwise equivalently processed, with a sample size of at least 5 for each comparison measurement condition.

[0141] As used herein, the term "about" means ±10%.

[0142] As used herein, the term "less than" means a lesser amount and an amount greater than zero.

[0143] The terms "comprise", "comprising", "includes", "including", "having" and their conjugates mean "including but not limited to".

[0144] The term "consisting of" means "including and limited to".

[0145] The term "consisting essentially of" means that a composition, method or structure may include additional ingredients, steps and / or parts, provided that the additional ingredients, steps and / or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

[0146] As used herein, the singular forms "a", "an", and "the" include plural references unless the context clearly indicates otherwise. For example, the term "a compound" or "at least one compound" may include multiple compounds, including mixtures thereof.

[0147] As used herein, the term "blood" refers to whole blood, leucocyte-reduced RBC, platelet-reduced RBC, and leucocyte- and platelet-reduced RBC. The term blood further includes packed red blood cells, platelet-reduced packed red blood cells, leucocyte-reduced packed red blood cells, and leucocyte- and platelet-reduced packed red blood cells. The temperature of the blood may vary according to the stage of the collection process, starting at the normal body temperature of 37°C at the time and point of collection, but once the blood leaves the patient's body, its temperature rapidly drops to about 30°C and then further drops to room temperature in about 6 hours when untreated and ultimately refrigerates at about 4°C to 6°C. Human red blood cells are in a dynamic state in the body. Red blood cells contain hemoglobin, an iron-containing protein that transports oxygen throughout the body and gives red blood its color. The percentage of blood volume composed of red blood cells is called the hematocrit. As used herein, unless otherwise restricted, RBC also includes packed red blood cells (pRBC). Packed red blood cells are prepared from whole blood using centrifugation techniques well known in the art. As used herein, unless otherwise stated, the hematocrit of pRBC is about 70%. As used herein, oxygen-reduced stored RBC may include oxygen- and carbon dioxide-reduced stored RBC. As used herein, oxygen-reduced (OR) blood may include oxygen- and carbon dioxide-reduced (OCR) blood.

[0148] As used herein, the terms "patient" and "subject" are used interchangeably to denote a human or animal in need of a blood transfusion.

[0149] As used herein, the term "trauma" includes bloodletting, hemorrhagic trauma.

[0150] As used herein, the term "hemorrhagic shock" is shock caused by the loss of circulating blood volume and / or oxygen-carrying capacity. Hemorrhagic shock is caused by any condition associated with blood loss, internal (e.g., gastrointestinal bleeding) or external bleeding, and trauma (e.g., penetrating or blunt trauma).

[0151] As used herein, the term "adverse event" includes events caused by hemorrhagic shock in patients with hemorrhagic trauma.

[0152] As used herein, the terms "injury", "damage", and "failure" refer to an organ not functioning properly or not functioning as expected in a human or animal without disease or injury.

[0153] As used herein, a "unit" of blood is about 450 - 500 ml and includes an anticoagulant. Suitable anticoagulants include citrate phosphate dextrose solution (CPD), citrate phosphate dextrose adenine (CPDA1), citrate dextrose (ACD), and anticoagulant citrate dextrose solution A (ACD-A).

[0154] Throughout this application, various aspects of the present disclosure may be presented in a range format. It should be understood that the description of a range format is merely for convenience and brevity and should not be construed as a rigid limitation on the scope of the present disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all possible sub-ranges as well as individual numerical values within that range. For example, a description of a range such as "1 to 6" should be considered to have specifically disclosed sub-ranges such as "1 to 3", "1 to 4", "1 to 5", "2 to 4", "2 to 6", "3 to 6", etc., as well as individual numbers within that range, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0155] Whenever a numerical range is indicated herein, it is intended to include any recited number (fractional or integral) within the indicated range. The phrases "ranging / ranges between a first recited number and a second recited number" and "ranging / ranges from a first recited number to a second recited number" are used interchangeably herein and are intended to include the first and second recited numbers and all fractional and integral numbers therebetween.

[0156] As used herein, the term "method" refers to a manner, means, technique, and procedure for accomplishing a given task, including, but not limited to, providing oxygen-reduced stored blood having an initial oxygen saturation of 20% or less and stored for at least 2 days to a human patient in need of a blood transfusion.

[0157] The present disclosure provides the following examples:

[0158] Example 1. A method for treating low mean arterial pressure in a subject in need thereof, the method comprising providing stored oxygen-reduced blood to a subject having low mean arterial pressure caused by hemorrhagic trauma, the oxygen-reduced blood having an initial oxygen saturation of 20% or less and maintained at an oxygen saturation of 20% or less for a storage period.

[0159] Example 2. The method according to Example 1, wherein after said providing, the mean arterial pressure of the subject in need thereof increases relative to a patient receiving conventionally stored blood.

[0160] Example 3. The method according to Example 1 or 2, wherein the hemorrhagic trauma is selected from the group consisting of surgery, penetrating trauma, blunt force trauma, fall injury, and motor vehicle crash injury.

[0161] Example 4. The method according to any one of Examples 1 to 3, wherein the rate of increase in the mean arterial pressure of the subject in need thereof is faster than the rate of increase in the mean arterial pressure of a subject not receiving the stored oxygen-reduced blood.

[0162] Example 5. The method according to any one of Examples 1 to 4, wherein after said provision, the mean arterial pressure of said subject in need increases by at least 20% relative to the mean arterial pressure of a patient receiving a transfusion of conventionally stored blood.

[0163] Example 6. The method according to any one of Examples 1 to 5, wherein after said provision, said increase in the mean arterial pressure of said subject is at least 20% and persists for at least 1 hour.

[0164] Example 7. The method according to any one of Examples 1 to 5, wherein said blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0165] Example 8. A method for reducing the amount of blood required for transfusion in a trauma patient in need, said method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0166] Example 9. The method according to Example 8, wherein said blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0167] Example 10. A method for reducing hemorrhagic shock in a trauma patient in need, said method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage, wherein said trauma patient has a lactate level of 0.5 mmol / L to 2.5 mmol / L before said provision, and wherein said hemorrhagic shock is reversed.

[0168] Example 11. The method according to Example 10, wherein said blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0169] Example 12. A method for reducing liver injury in a trauma patient in need of blood transfusion therapy, said method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0170] Example 13. A method according to Example 9, wherein said trauma patient has an improved aspartate aminotransferase (AST) level, alanine aminotransferase (ALT) level, or a combination thereof after said provision.

[0171] Example 14. The method according to Example 13, wherein said blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0172] Example 15. A method for reducing renal failure in a trauma patient in need of blood transfusion therapy, said method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0173] Example 16. The method according to Example 15, wherein the trauma patient has an improved level selected from neutrophil gelatinase-associated lipocalin (NGAL), serum creatinine, blood urea nitrogen (BUN), or a combination thereof.

[0174] Example 17. The method according to Example 15 or 16, wherein the blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0175] Example 18. A method for reducing lung injury in a trauma patient in need of blood transfusion therapy, the method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0176] Example 19. The method according to Example 18, wherein the blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0177] Example 20. A method for reducing lactic acid in a trauma patient in need thereof, the method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0178] Example 21. The method according to Example 20, wherein the blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0179] Example 22. A method for reducing aspartate aminotransferase (AST) in a trauma patient in need thereof, the method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0180] Example 23. A method for reducing alanine aminotransferase (ALT) in a trauma patient in need thereof, the method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0181] Example 24. A method for reducing the blood urea nitrogen (BUN) level in a trauma patient in need thereof, the method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0182] Example 25. A method for reducing neutrophil gelatinase-associated lipocalin (NGAL) in a trauma patient in need thereof, the method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0183] Example 26. A method for reducing serum creatinine in a trauma patient in need thereof, the method comprising providing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0184] Example 27. The method according to any one of Examples 22 to 26, wherein the blood is stored blood with reduced oxygen and reduced carbon dioxide.

[0185] Example 28. Use of donor blood in the manufacture of oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage for therapeutic applications in low mean arterial pressure.

[0186] Example 29. Use of donor blood in the manufacture of oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage for therapeutic applications to reduce adverse events in patients with hemorrhagic trauma, wherein the adverse events are selected from the group consisting of liver injury, lung failure, kidney failure, and heart failure.

[0187] Example 30. The use according to Example 28 or 29, wherein the manufacture comprises blood with reduced oxygen and carbon dioxide.

[0188] Example 31. A method of treating a patient in need with stored oxygen-reduced blood having an oxygen saturation of 20% or less during storage, wherein the patient in need has an inflammation.

[0189] Example 32. The method according to Example 31, wherein the inflammation is chronic or acute.

[0190] Example 33. The method according to Example 31, wherein the inflammation is systemic.

[0191] Example 34. The method according to any one of Examples 31 to 33, wherein the inflammation is caused by trauma, infection, cancer, coagulopathy, or autoimmunity.

[0192] Example 35. The method according to any one of Examples 31 to 34, wherein the patient is further accompanied by fever or at risk of developing fever.

[0193] Example 36. The method according to Example 35, wherein the fever is at least 38 °C.

[0194] Example 37. The method according to any one of Examples 31 to 36, wherein the patient has an increased C-reactive protein (CRP) level.

[0195] Example 38. The method according to Example 37, wherein the CRP level of the patient is at least 2.0 mg / L.

[0196] Example 39. The method according to Example 38, wherein the CRP level of the patient is at least 3.0 mg / L.

[0197] Example 40. The method according to any one of Examples 37 to 39, wherein the CRP level of the patient is 3.0 mg / L to 10 mg / L.

[0198] Example 41. The method according to any one of Examples 31 to 40, wherein the levels of one or more inflammatory mediators selected from the group consisting of IL-6, IL-8, IL-10, and granulocyte colony-stimulating factor (G-CSF) in the patient are increased compared to a patient without fever.

[0199] Example 42. The method according to Example 41, wherein the IL-6 level of the patient is at least 5 pg / mL.

[0200] Example 43. The method according to Example 41, wherein the IL-8 level of the patient is at least 29 pg / mL.

[0201] Example 44. The method according to Example 41, wherein the IL-10 level of the patient is at least 15 pg / mL.

[0202] Example 45. The method according to Example 41, wherein the G-CSF level of the patient is at least 35 pg / mL.

[0203] Example 46. The method according to any one of Examples 31 to 45, wherein the levels of one or more inflammatory mediators selected from the group consisting of CCL5 and CXCL10 in the patient are decreased compared to a patient without fever.

[0204] Example 47. The method according to Example 46, wherein the CCL5 level of the patient is at least 380 pg / mL.

[0205] Example 48. The method according to Example 46, wherein the CXCL10 level of the patient is at least 270 pg / mL.

[0206] Example 49. The method according to any one of Examples 31 to 48, wherein the patient has liver damage.

[0207] Example 50. The method according to any one of Examples 31 to 49, wherein the patient has lung damage.

[0208] Example 51. The method according to any one of Examples 31 to 50, wherein the patient has spleen damage.

[0209] Example 52. The method according to any one of Examples 31 to 51, wherein the patient has kidney damage.

[0210] Example 53. The method according to any one of Examples 31 to 52, wherein the patient has a bone lesion.

[0211] Example 54. The method according to any one of Examples 31 to 53, wherein the treatment reduces the adhesion of red blood cells to thrombospondin in the patient as compared to a patient treated with conventionally stored blood.

[0212] Example 55. The method according to Example 54, wherein the thrombospondin is reduced by at least 10%.

[0213] Example 56. The method according to any one of Examples 31 to 55, wherein the stored oxygen-reduced blood is blood reduced in oxygen and carbon dioxide.

[0214] Example 57. A method of improving blood transfusion in a sickle cell patient in need thereof, the method comprising providing stored oxygen-reduced blood to a patient suffering from sickle cell disease, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage.

[0215] Example 58. The method according to Example 57, wherein the sickle cell disease is sickle cell anemia.

[0216] Example 59. The method according to Example 57, wherein the sickle cell disease is sickle cell crisis.

[0217] Example 60. The method according to Example 57, wherein the sickle cell disease is selected from the group consisting of hemoglobin SS (HbSS), hemoglobin SC (HbSC), hemoglobin Sβ thalassemia + (HbSB+), hemoglobin S(β-0) thalassemia (HbSB), hemoglobin SD (HbSD), hemoglobin SE (HbSE), and hemoglobin SO (HbSO).

[0218] Example 61. The method according to any one of Examples 57 to 60, wherein the improvement comprises reducing the occurrence of dactylitis (hand-foot syndrome) by at least 10% as compared to a patient receiving conventionally stored blood.

[0219] Example 62. The method according to any one of Examples 57 to 61, wherein the improvement comprises reducing the number of pain crises by at least 10% as compared to treatment with conventionally stored blood.

[0220] Example 63. The method according to any one of Examples 57 to 62, wherein the improvement comprises reducing complications due to anemia, the complications being selected from the group consisting of fatigue, irritability, dizziness, dyspnea, pallor, jaundice, slow growth, and developmental delay.

[0221] Example 64. The method according to any one of Examples 57 to 63, wherein the improvement comprises reducing the occurrence of infection.

[0222] Example 65. The method according to any one of Examples 57 to 64, wherein the improvement comprises reducing spleen damage.

[0223] Example 66. The method according to Example 65, wherein the spleen damage is autosplenism.

[0224] Example 67. The method according to Example 65, wherein the spleen damage is splenomegaly.

[0225] Example 68. The method according to any one of Examples 57 to 67, wherein the improvement comprises reducing or preventing stroke.

[0226] Example 69. The method according to any one of Examples 57 to 68, wherein after the blood transfusion, the previously stored oxygen-reduced blood has a higher 24-hour recovery compared to conventionally stored blood.

[0227] Example 70. The method according to any one of Examples 57 to 69, wherein the previously stored oxygen-reduced blood has increased deformability compared to conventionally stored blood.

[0228] Example 71. The method according to any one of Examples 57 to 70, wherein the oxygen-reduced blood has increased deformability compared to conventionally stored blood and when sickle cell plasma is present.

[0229] Example 72. The method according to any one of Examples 57 to 71, further comprising reducing carbon dioxide in the oxygen-reduced blood.

[0230] Example 73. A method of reducing the number of vaso-occlusive episodes in a patient in need thereof, the method comprising providing stored oxygen-reduced blood to a patient in need thereof, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage, and wherein reducing the vaso-occlusive episodes comprises reducing the adhesion of red blood cells to endothelial cells.

[0231] Example 74. The method according to Example 73, wherein the patient in need thereof has sickle cell disease.

[0232] Example 75. The method according to Example 73 or 74, wherein the patient in need thereof has inflammation.

[0233] Example 76. The method according to any one of Examples 73 to 75, wherein said reducing red blood cell adhesion is reducing adhesion to endothelial cells expressing thrombospondin.

[0234] Example 77. The method according to any one of Examples 73 to 76, wherein said reducing red blood cell adhesion is reducing adhesion to endothelial cells expressing vascular cell adhesion molecule-1 (VCAM-1).

[0235] Example 78. The method according to any one of Examples 73 to 77, wherein said reducing red blood cell adhesion is reducing adhesion to endothelial cells expressing laminin.

[0236] Example 79. The method according to any one of Examples 76 to 78, wherein said adhesion is reduced by at least 10% compared to conventionally stored blood.

[0237] Example 80. The method according to any one of Examples 76 to 79, wherein said adhesion is reduced by at least 20% compared to conventionally stored blood.

[0238] Example 81. The method according to any one of Examples 76 to 80, wherein said adhesion is reduced by 10% to 50%.

[0239] Example 82. The method according to any one of Examples 73 to 81, wherein said sickle cell disease is sickle cell anemia.

[0240] Example 83. The method according to any one of Examples 73 to 81, wherein said sickle cell disease is sickle cell crisis.

[0241] Example 84. The method according to any one of Examples 73 to 83, wherein said sickle cell disease is selected from the group consisting of hemoglobin SS (HbSS), hemoglobin SC (HbSC), hemoglobin Sβ-thalassemia+ (HbSB+), hemoglobin S (β-0) thalassemia (HbSB), hemoglobin SD (HbSD), hemoglobin SE (HbSE), and hemoglobin SO (HbSO).

[0242] Example 85. The method according to any one of Examples 73 to 83, further comprising reducing carbon dioxide in said oxygen-reduced blood.

[0243] Example 86. A method of reducing red blood cell adhesion to endothelial cells in a patient in need thereof, said method comprising providing stored oxygen-reduced blood to a patient suffering from sickle cell disease, wherein said oxygen-reduced blood has an oxygen saturation of 20% or less during storage.

[0244] Example 87. Oxygen-reduced blood having an oxygen saturation of 20% or less during storage for treating sickle cell disease.

[0245] Example 88. The oxygen-reduced blood according to Example 87, wherein the blood is oxygen- and carbon dioxide-reduced blood.

[0246] Example 89. Oxygen-reduced blood having an oxygen saturation of 20% or less during storage for treating inflammation.

[0247] Example 90. The oxygen-reduced blood according to Example 89, wherein the blood is oxygen- and carbon dioxide-reduced blood.

[0248] Example 91. Stored oxygen-reduced blood having an oxygen saturation of 20% or less for treating vaso-occlusive crises, wherein the treatment reduces the number of vaso-occlusive crises in a patient in need thereof, and the reducing of vaso-occlusive crises includes reducing the adhesion of red blood cells to endothelial cells.

[0249] Example 92. The stored oxygen-reduced blood according to Example 91, wherein the blood is stored oxygen- and carbon dioxide-reduced blood.

[0250] Example 93. Use of oxygen-reduced blood having an oxygen saturation of 20% or less during storage for treating inflammation in a patient in need thereof.

[0251] Example 94. Use of oxygen-reduced blood having an oxygen saturation of 20% or less during storage for treating sickle cells in a patient in need thereof.

[0252] Example 95. Use of the oxygen-reduced blood according to Example 94, wherein the blood is oxygen- and carbon dioxide-reduced blood.

[0253] Example 96. Use of oxygen-reduced blood substantially as shown and described.

[0254] Example 97. Use of oxygen- and carbon dioxide-reduced blood substantially as shown and described.

[0255] Example 98. A kit comprising an oxygen-impermeable blood storage bag, a filter, and a drip set, the blood storage bag containing stored oxygen-reduced blood having an oxygen saturation of 20% or less during storage.

[0256] Example 99. The kit according to Example 98, further comprising means for adding gas to the stored oxygen-reduced blood prior to transfusion.

[0257] Example 100. The kit according to Example 98 or 99, further comprising a conduit for infusing the stored oxygen-reduced blood into a patient in need thereof.

[0258] Example 101. Use of the kit according to any one of Examples 98 to 100 in the treatment of sickle cell disease or inflammation in a patient in need thereof.

[0259] Although the present disclosure has been described with reference to specific embodiments, those skilled in the art will understand that various changes can be made and elements thereof can be replaced with equivalents without departing from the scope of the present disclosure. Additionally, various modifications can be made to adapt a particular situation or material to the teachings of the present disclosure without departing from the scope of the present disclosure.

[0260] Therefore, the present disclosure is not intended to be limited to the specific embodiments disclosed as the best mode contemplated for carrying out the present disclosure, but rather the present disclosure will include all embodiments falling within the scope and spirit of the appended claims.

[0261] Examples

[0262] Example 1: Blood collection and sample preparation

[0263] Each red blood cell pool was collected from a total of 12 - 14 CP2D anticoagulant rats. The collected blood was leukoreduced using a neonatal leukoreduction filter, the components were separated and the RBCs were stored in AS-3 additive solution. A total of two RBC pools were collected. Each pool was divided into four types: untreated control (C), sham control (SC), oxygen-reduced (OR), and oxygen and carbon dioxide-reduced (OCR). For the C, SC, OR, and OCR units, the RBC subunits were processed by transferring them into 80 mL PVC blood transfer bags, and the final RBC products were prepared by a gas exchange process. The RBC bags other than C were filled with 100% N2 (for OR) or 95% N2 / 5% CO2 (for OCR) or air (SC) through a sterile filter and slowly rotated at 2 RPM - 3 RPM on their long sides (except for C). For the OR and OCR units, after 10 minutes, the gas was removed through the filter and fresh gas was introduced for subsequent gas exchange processes. This process was repeated 5 to 8 times until the target %SO2 of 5% - 10% was reached, as measured by an ABL-90 blood gas analyzer (Radiometer Copenhagen). The SC unit was rotated for 60 minutes without any gas exchange. The OR and OCR units were anaerobically stored in N2-filled tanks, while the C and SC units were stored in ambient air. All units were stored at 4°C for 3 weeks and sampled at day 0 or 1, 7, 14, 21, and 28. Two pools were prepared and stored.

[0264] ATP, 2,3-DPG, and hemolysis analyses were performed on days 0, 1, 7, 14, 21, and 28. As Figure 1 shown, ATP levels were higher in OR blood on day 21 and in OCR blood on days 7, 14, 21, and 28 compared to conventionally stored blood (control). OR blood also had higher 2,3-DPG levels on days 2, 7, and 14 compared to the control. OCR blood also showed higher 2,3-DPG levels on days 2, 7, and 14 compared to the control. See Figure 2 .

[0265] Example 2: Recovery of Oxygen-Deficient Blood

[0266] Small volumes (less than 200 μL) of control, OR, and OCR blood stored for 3 weeks were labeled with technetium-99m. The labeled RBCs (less than 200 uL) were infused into animals and the circulating radioactivity was measured periodically up to 24 hours to estimate the proportion of infused RBCs surviving at 24 hours post-infusion. As Figure 3 shown, significantly more OR- and OCR-RBCs were recovered compared to control RBCs when the RBCs were stored for three weeks.

[0267] Example 3: Rat Model of Hemorrhagic Shock Resuscitation

[0268] Blood collection and sample preparation: Each erythrocyte pool was collected from a total of 12 - 14 CP2D anticoagulant rats. The collected blood was leukoreduced using a neonatal leukoreduction filter, the components were separated and the RBCs were stored in AS-3 additive solution. A total of 6 RBC pools were collected. Two pools were prepared for conventional storage (control). Two pools had no oxygen (oxygen-deficient; OR), and the remaining two blood pools had no oxygen and carbon dioxide (oxygen- and carbon dioxide-deficient; OCR). Each of the four pools to be reduced was processed by transferring the RBCs into a 600 mL PVC blood transfer bag, and the final RBC product was prepared by a gas exchange process. The RBC bags were filled with 100% N2 (for OR) or 95% N2 / 5% CO2 (for OCR) through a sterile filter and slowly rotated at 60 RPM - 90 RPM on their long sides. After 10 minutes, the gas was removed through the filter and fresh gas was introduced for subsequent gas exchange processes. This process was repeated 5 to 8 times until the target %SO2 of 5% - 10% was reached, as measured by an ABL-90 blood gas analyzer (Radiometer Copenhagen). The OR and OCR blood was stored anaerobically in N2-filled tanks.

[0269] Studies were conducted in Sprague - Dawley rats (Charles River Laboratories, Boston, MA) weighing 150 - 200 grams (g). Briefly, animals were anesthetized by intraperitoneal administration of 40 mg / kg sodium pentobarbital. The animals were placed in the supine position on a heating pad to maintain core body temperature at 37°C. The animals were prepared as follows: (i) left jugular vein and left femoral artery catheterization; (ii) tracheotomy (polyethylene - 90 tube); and (iii) introduction of a left ventricular (LV) conductance catheter through the right carotid artery. The animals were mechanically ventilated with room air (TOPO ventilator, Kent Scientific, Torrington, CT) at a respiratory rate of 50 - 70 breaths per minute and a peak inspiratory pressure of 10 cmH2O - 15 cmH2O. After instrumentation, inhaled anesthesia (1.5% / vol isoflurane, Dragerwerk AG, Laubeck, Germany) was administered using a vaporizer connected to the ventilator. The depth of anesthesia was continuously verified by toe - pinching and, if necessary, isoflurane was increased by 0.1% / vol to prevent animal discomfort.

[0270] The anesthetized animals were bled by withdrawing 50% of the animal blood volume (BV; estimated as 7% of body weight) via the femoral artery catheter over 10 minutes, putting the animals in a state of hypovolemic shock. The hypovolemic shock state was maintained for 30 minutes. Resuscitation was performed by infusing previously stored RBC at 300 microliters per minute (μL / min) via the femoral artery until the mean arterial pressure (MAP) stabilized at 90% of the baseline during a 60 - minute resuscitation period. At 10, 20, 30, 45, and 60 minutes during this period, MAP and heart rate (HR) were obtained from the femoral artery catheter (PowerLab, AD Instruments, Colorado Springs, CO). Sixty minutes later, hematocrit (Hct) was measured by centrifugation of heparinized capillary tubes. Hemoglobin (Hb), lactate, glucose, K+, Na+, pH, and arterial blood gases were determined by an ABL90 blood gas analyzer (Radiometer, Copenhagen). Indicators of cardiac function and systemic values (MAP, HR, Hct, Hb, and blood gases) were monitored at baseline (BL), during shock, and at 10 (early R), 20, 30, 45, and 60 (late R) minutes after resuscitation. The animals were euthanized at the end of the experiment.

[0271] Example 4: Hematocrit analysis in a hemorrhagic shock rat model

[0272] After induction of hypovolemic shock, the hematocrit (Hct) decreased by approximately 30% to 40%. Providing conventional, OR, or OCR blood stored for 1 week was able to restore the hematocrit to normal levels. SeeFigure 4A However, as Figure 4B shown, after 10 minutes of resuscitation (early R), compared with control and OCR blood, OR blood stored for 1 week showed an increased percentage of hematocrit. At 60 minutes after resuscitation (late R), the percentage of hematocrit in OR blood was still improved compared with the control.

[0273] Example 5: Mean arterial pressure changes with oxygen-reduced blood

[0274] Mean arterial pressure (MAP) was obtained from a femoral artery catheter (PowerLab, ADInstruments, Colorado Springs, CO). As Figure 5A shown, the baseline MAP was 80 mmHg to 110 mmHg. During hemorrhagic shock, the MAP decreased to 20 mmHg to 60 mmHg. Resuscitation of the animals with OR and OCR blood stored for one week increased the MAP to approximately 80 mmHg and 90 mmHg, respectively. As Figure 5B shown, compared with the control, resuscitation with OR blood was able to restore the MAP to the normal range after 10 minutes. After 60 minutes of resuscitation, the control and OCR-stored blood were able to restore the MAP to the normal range. The amount of blood required for resuscitation and maintenance of hemodynamics using conventionally stored RBCs (control) was greater than that required for OR and OCR RBCs. See Figure 6A and Figure 6B .

[0275] Example 6: Metabolic response to hemorrhagic shock

[0276] Hemorrhagic shock in the animals increased the lactate level from approximately 2 mmol / L to approximately 8 mmol / L to 14 mmol / L. After only 10 minutes of resuscitation, resuscitation with OR and OCR RBCs stored for one week decreased the lactate level to near normal levels. See Figure 7A . The lactate level in the animals resuscitated with control blood was similar to that in the hemorrhagic shock animals. Animals treated with control, OR, and OCR RBCs for 60 minutes showed similar lactate levels. As Figure 7B shown, after 10 minutes of resuscitation, compared with the control, OR RBCs stored for 3 weeks were also able to decrease the lactate level. However, after 80 minutes of resuscitation, OCR RBCs restored the lactate level to the normal range. Control and OR RBCs were able to decrease the lactate level but not to the normal range of 1 mmol / L to 3 mmol / L. Analysis of glucose levels showed that in the animals with hemorrhagic shock, the normal range of approximately 160 mg / dL to approximately 240 mg / dL of glucose increased to the range of approximately 320 mg / dL to approximately 510 mg / dL. See Figure 8A and Figure 8B。After 10 minutes of resuscitation, compared with the control, both OR and OCR RBC stored for one week decreased the glucose level. After 60 minutes of resuscitation, all three samples restored the glucose level to the normal range. As Figure 8B shown, after 10 minutes of resuscitation, compared with the control, OR and OCR RBC stored for three weeks were also able to decrease the glucose level. Different from the RBC stored for one week, only OR and OCR RBC were able to restore the glucose to the normal range. Therefore, compared with the control RBC, OR and OCR RBC decreased the lactate and glucose levels faster.

[0277] Example 7: Injury and inflammation of vital organs

[0278] Analyze the organ injury and inflammation in animals after hemorrhagic shock and resuscitation. Elevated liver enzyme levels indicate some form of liver damage or injury. Analyze the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) to determine liver damage. Compared with the control RBC stored for the same period, resuscitation with OR and OCR RBC stored for one week previously ( Figure 9A and Figure 10A ) and three weeks ( Figure 9B and Figure 10B ) decreased the AST and ALT levels. Analyze the serum creatinine and blood urea nitrogen (BUN) levels to determine renal function. Compared with the control RBC, OR and OCR RBC stored for one week decreased the serum creatinine level by more than 30% ( Figure 11A ). After three weeks of storage, the serum creatinine levels of the animals treated with control, OR, and OCR RBC overlapped ( Figure 11B ). Compared with the control, in the animals treated with OCR RBC stored for one week, the BUN level decreased by more than 30% ( Figure 12A ). Compared with the control, in the animals treated with OR RBC stored for three weeks, the BUN level also decreased by more than 30% ( Figure 12B ). Overall, compared with the control RBC, OR and OCR RBC preserved the vital organ function.

[0279] After the in vivo study was completed, the liver, spleen, and lungs were excised from the animals, and various inflammatory factors, including CXCL1, urinary neutrophil gelatinase-associated lipocalin (u-NGAL), IL-6, and neutrophils, were analyzed. Compared with the control stored for the same period, CXCL1 decreased in the animals treated with OR and OCR RBC stored for one week or three weeks ( Figure 13A , 13B , Figure 14A , 14B and Figure 15A , 15B ). As Figure 16A and 16BAs shown, compared with control RBCs stored for an equal amount of time, u-NGAL was decreased in the kidneys of animals treated with OR or OCR RBCs stored for one or three weeks ( Figure 16A and 16B ). As Figure 17A , 17B and Figures 18A, 18B show, compared with control RBCs stored for the same period of time, OR and OCR RBCs significantly reduced the percentage of CD45+ neutrophils and IL-6 levels in the lungs excised from animals. These results indicate that organ damage and inflammation were reduced in animals treated with OR and OCR RBCs compared with those treated with control RBCs.

[0280] Example 8: Thrombospondin Adhesion Assay

[0281] Thrombospondin is an important biomarker of disease severity in sickle cell disease. During vaso-occlusive crises (episodes) of sickle cell disease, circulating thrombospondin levels increase significantly, and at the same time, the adhesion of sickle red blood cells to the vascular endothelium increases correspondingly. Conventional stored RBCs and oxygen-reduced RBCs were compared to determine the differences in adhesion to endothelial cells. Briefly, two units of O-positive whole blood were combined in citrate-phosphate-dextrose (CPD) / SAGM. Each unit was filtered through a leukoreduction filter at room temperature. The leukoreduced whole blood (LR-WB) was transferred to a 2-liter blood collection bag. Then the combined LR-WB was divided into equal 500-ml aliquots (Unit A and Unit B) and processed into packed red blood cells and plasma. Plasma was removed and stored at -70 °C. The packed RBCs (pRBCs) in Unit A were resuspended in SAGM, and the pRBCs in Unit B were resuspended in PAGGSM. The pRBCs in Unit B were deoxygenated for 3 hours at room temperature using the Hemanext oxygen reduction system. The units were stored at 4 °C for 42 days. After pre-incubation in normal plasma or plasma from sickle cell disease patients, samples were taken from each unit and analyzed to determine the complete blood count (CBC), gas group analysis, and the adhesion of thrombospondin on the microchip. Samples collected on Day 0 were incubated with 0.4% bovine serum albumin (BSA), normal plasma (AA), plasma from sickle cell patients in steady state (SS), or plasma from sickle cell patients in crisis. Then the incubated samples were incubated with the adhesion assay microchip. To determine the effect of different shear stresses on the adhesion of oxygen-reduced RBCs and conventionally stored RBCs, the cells were subjected to an increasing shear stress from 0.5 dyne / cm 2 to 5 dyne / cm 2 . As Figures 19A to 19CAs shown, the system is stable and responds to changes in shear stress. As the shear stress increases, the number of cells adhering to the thrombospondin matrix gradually decreases. These results further demonstrate the adhesion strength of RBCs to thrombospondin.

[0282] To test the effect of RBC storage conditions on thrombospondin adhesion, samples collected on days 0, 21, and 42 were subjected to shear stresses of 0.5 dyn / cm 2 ( Figure 20A )、2.0 dyn / cm 2 ( Figure 20B ) or 5.0 dyn / cm 2 ( Figure 20C ). As Figures 20A to 20C shown, compared to conventionally stored red blood cells, oxygen-depleted red blood cells have significantly less adhesion to thrombospondin.

[0283] Example 9: Hemolysis of Oxygen-Depleted and Control RBCs

[0284] To determine the effect of sickle plasma on hemolysis, 20 mL samples were collected from two sickle cell disease patients and one healthy patient (HT) more than 28 days after exchange transfusion (TF). The samples were dispensed into 15 mL centrifuge tubes and centrifuged at 1500 x g for 15 minutes at room temperature (without braking). Then, the platelet-rich plasma (PRP) from each 15 mL tube was transferred to a new 15 mL tube and centrifuged at 1500 x g for 15 minutes at room temperature (without braking). The PRP was transferred from each centrifuge tube to a 15 mL combined tube using a clean disposable pipette. The platelet-poor plasma (PPP) was aliquoted into autoclaved and pre-labeled 1.5 mL tubes, 1 mL per tube. The PPP was stored at -40 °C until use. Flow adhesion was performed with RBCs resuspended in the two TF samples. The TF PPP sample (TF006) Figure 21 ) that supported higher baseline adhesion was selected.

[0285] Three groups of pRBCs were prepared and stored: conventionally stored (A), carbon dioxide-depleted and 40% oxygen (B), and carbon dioxide and oxygen-depleted (C). Aliquots were collected from each group on days 1 and 21 of storage. The samples were transferred to 15 mL centrifuge tubes and centrifuged at 1500 x g for 15 minutes. The supernatant was discarded, and the RBC pellet was reconstituted. 80 μL of packed and washed RBCs from A, B, and C were resuspended in 40 μL of prepared PPP from sickle cell plasma (SS) and healthy plasma (HT) and incubated at 37 °C on a nutator (4 RPM) for 2 hours and 24 hours. Compared to conventionally stored RBCs, carbon dioxide and oxygen-depleted RBCs reduced the percentage of hemolysis.

[0286] Example 10: Effect of Sickle Plasma on Deformability

[0287] Three RBC units were combined, leukoreduced (LR), and divided equally into Group A (control) and Group B (oxygen-reduced). The Group B RBCs were treated to contain approximately 5% SO and approximately 8 mmHg CO2. The Group A and Group B RBCs were stored at 42 °C for 42 days. Aliquots of RBCs from Group A and Group B were incubated at 37 °C with (1) healthy donor plasma for 3 hours, (2) with healthy donor plasma for 12 hours, (2) with sickle cell donor plasma for 3 hours, and (4) with sickle cell donor plasma for 12 hours. Matched samples from Group A and Group B were analyzed in parallel on a microvascular analysis chip. Deformability was measured as total exit flow. Oxygen-reduced RBCs incubated with sickle cell plasma had increased deformability compared to control RBCs incubated with healthy plasma. This increase in deformability was observed after 3 and 12 hours of incubation.

[0288] The following corresponds to the original claims in the parent application and is hereby incorporated herein as part of the specification:

[0289] 1. A method of treating a patient in need thereof with stored oxygen-reduced blood having an oxygen saturation of 20% or less during storage, wherein the patient in need thereof has an inflammation.

[0290] 2. The method according to item 1, further comprising reducing carbon dioxide in the oxygen-reduced blood.

[0291] 3. The method according to item 1, wherein the inflammation is caused by trauma, infection, cancer, coagulopathy, or autoimmunity.

[0292] 4. The method according to item 1, wherein the patient further has a fever or is at risk of developing a fever.

[0293] 5. The method according to item 1, wherein the patient has an increased C-reactive protein (CRP) level.

[0294] 6. The method according to item 1, wherein the levels of one or more inflammatory mediators selected from the group consisting of IL-6, IL-8, IL-10, and granulocyte colony-stimulating factor (G-CSF) are increased in the patient compared to a patient without a fever.

[0295] 7. The method according to item 1, wherein the levels of one or more inflammatory mediators selected from the group consisting of CCL5 and CXCL10 are decreased in the patient compared to a patient without a fever.

[0296] 8. The method according to item 1, wherein the patient has liver damage, lung damage, spleen damage, kidney damage, bone damage, or a combination thereof.

[0297] 9. The method according to item 1, wherein the treatment reduces the adhesion of red blood cells to thrombospondin in the patient as compared to a patient treated with conventionally stored blood.

[0298] 10. A method of improving blood transfusion in a sickle cell patient in need thereof, the method comprising providing stored oxygen-reduced blood to a patient with sickle cell disease, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage.

[0299] 11. The method according to item 10, wherein the improvement comprises reducing the occurrence of dactylitis (hand-foot syndrome) by at least 10% as compared to a patient receiving conventionally stored blood.

[0300] 12. The method according to item 10, wherein the improvement comprises reducing the number of pain crises by at least 10% as compared to treatment with conventionally stored blood.

[0301] 13. The method according to item 10, wherein the improvement comprises reducing complications due to anemia selected from the group consisting of fatigue, irritability, dizziness, dyspnea, pallor, jaundice, slow growth, and developmental delay.

[0302] 14. The method according to item 10, wherein the improvement comprises reducing the occurrence of infections.

[0303] 15. The method according to item 10, wherein the improvement comprises reducing spleen damage.

[0304] 16. The method according to item 10, wherein the improvement comprises reducing or preventing stroke.

[0305] 17. A method of reducing the number of vaso-occlusive episodes in a patient in need thereof, the method comprising providing stored oxygen-reduced blood to a patient in need thereof, wherein the oxygen-reduced blood has an oxygen saturation of 20% or less during storage, and wherein reducing the vaso-occlusive episodes comprises reducing the adhesion of red blood cells to endothelial cells.

[0306] 18. The method according to item 17, further comprising reducing carbon dioxide in the oxygen-reduced blood.

[0307] 19. The method according to item 17, wherein the patient in need thereof has sickle cell disease, inflammation, or sickle cell disease and inflammation.

[0308] 20. A method for reducing the adhesion of red blood cells to endothelial cells in a patient in need, the method comprising providing stored oxygen-depleted blood to a patient suffering from sickle cell disease, wherein the oxygen-depleted blood has an oxygen saturation of 20% or less during storage.

Claims

1. Use of a stored oxygen-reduced blood product having an oxygen saturation of 20% or less during storage for the manufacture of a medicament for treating elevated C-reactive protein (CRP) levels in a patient in need suffering from inflammation, wherein treating elevated CRP in said patient in need comprises administering said medicament to said patient in need, wherein said patient in need has an elevated CRP level of 5 milligrams per liter (mg / L) to 10 mg / L prior to said administration, wherein said patient in need has a fever or is at risk of developing a fever, and wherein said administration reduces the elevated CRP level in said patient in need by at least 20% as compared to the CRP level of a patient suffering from inflammation who has been administered a medicament manufactured from a non-oxygen-reduced conventionally stored blood product.

2. The use according to claim 1, wherein said stored oxygen-reduced blood further comprises reduced carbon dioxide.

3. The use according to claim 1, wherein said inflammation is caused by trauma, infection, cancer, coagulopathy or autoimmunity.

4. The use according to claim 1, wherein said administration reduces the elevated CRP level in said patient in need by at least 40% as compared to the CRP level of a patient suffering from inflammation who has been administered a non-oxygen-reduced conventionally stored blood product.

5. The use according to claim 1, wherein the level of one or more inflammatory mediators selected from the group consisting of interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10) and granulocyte colony-stimulating factor (G-CSF) is increased in said patient in need as compared to a patient suffering from inflammation but not having a fever or not being at risk of developing a fever.

6. The use according to claim 1, wherein the level of one or more inflammatory mediators selected from the group consisting of chemokine ligand 5 (CCL5) and CXC-motif chemokine ligand 10 (CXCL10) is decreased in said patient in need as compared to a patient suffering from inflammation but not having a fever or not being at risk of developing a fever.

7. The use according to claim 1, wherein said patient in need has liver damage, lung damage, spleen damage, kidney damage, bone damage or any combination thereof.

8. The use according to claim 1, wherein said administration reduces the adhesion of red blood cells to thrombospondin in said patient in need by at least 10% as compared to a patient suffering from inflammation and administered a non-oxygen-reduced conventionally stored blood product.

9. The use according to claim 1, wherein said inflammation is chronic inflammation, acute inflammation or systemic inflammation.

10. The use according to claim 1, wherein after said administration the elevated CRP level is reduced to less than 3 mg / L.

11. The use according to claim 5, wherein the elevated IL-6 level is at least 5 picograms per milliliter (pg / mL); the elevated IL-8 level is at least 29 picograms per milliliter (pg / mL); The elevated IL-10 level is at least 15 picograms per milliliter (pg / mL); The elevated G-CSF level is at least 35 picograms per milliliter (pg / mL); or Any combination thereof.

12. The use according to claim 6, wherein The reduced CCL5 level is at least 380 picograms per milliliter (pg / mL); The reduced CXCL10 level is at least 270 picograms per milliliter (pg / mL); or Any combination thereof.

13. The use according to claim 5 or claim 11, wherein relative to a patient with inflammation but without fever or not at risk of developing fever and who has received a non-oxygen-reduced conventionally stored blood product, the administration reduces the level of one or more elevated inflammatory mediators by at least 20%.

14. The use according to claim 6 or claim 12, wherein relative to a patient with inflammation but without fever or not at risk of developing fever and who has received a non-oxygen-reduced conventionally stored blood product, the administration increases the level of one or more reduced inflammatory mediators by at least 20%.

15. The use according to claim 1, wherein relative to a patient with inflammation who has received a drug manufactured from non-oxygen-reduced conventionally stored blood, the administration reduces the adhesion of red blood cells to thrombospondin by at least 10%.

16. Use of stored oxygen-reduced blood having an oxygen saturation of 20% or less during storage for the manufacture of a drug for reducing or preventing stroke in a patient with sickle cell disease in need thereof, wherein reducing or preventing stroke comprises administering the drug to the patient in need thereof, wherein when sickle cell plasma is present, the stored oxygen-reduced blood has increased deformability compared to non-oxygen-reduced conventionally stored blood, wherein the increased deformability is determined by microvascular chip analysis, and wherein reducing or preventing stroke further comprises reducing the number of pain crises by at least 10% in the patient in need thereof compared to a patient with sickle cell disease who has received a drug manufactured from non-oxygen-reduced conventionally stored blood after the administration.

17. The use according to claim 16, wherein reducing or preventing stroke further comprises reducing the number of occurrences of dactylitis (hand-foot syndrome) by at least 10% compared to a patient with sickle cell disease who has received a drug manufactured from non-oxygen-reduced conventionally stored blood.

18. The use according to claim 16 or claim 17, wherein Reducing or preventing stroke further comprises reducing the number of complications caused by anemia in the patient with sickle cell disease in need thereof after administration, wherein the complications caused by anemia are selected from the group consisting of fatigue, irritability, dizziness, dyspnea, pallor, jaundice, slow growth, developmental delay, and any combination thereof; or Reducing or preventing stroke further comprises reducing the occurrence of infection, spleen damage, or both in the patient with sickle cell disease in need thereof after administration. Use of stored oxygen-depleted blood having an oxygen saturation of 20% or less during storage for the manufacture of a medicament for reducing the occurrence of vaso-occlusive episodes in a patient in need thereof, wherein reducing the occurrence of vaso-occlusive episodes comprises administering the medicament to the patient in need thereof, and wherein the administration reduces the adhesion of red blood cells to thrombospondin by at least 10% in the patient in need thereof relative to a patient to whom a medicament made from non-oxygen-depleted conventionally stored blood has been administered. Use of stored oxygen-depleted blood having an oxygen saturation of 20% or less during storage for the manufacture of a medicament for reducing the adhesion of red blood cells to endothelial cells expressing thrombospondin, vascular cell adhesion molecule-1 (VCAM-1), or laminin in a patient in need thereof, wherein reducing the adhesion of red blood cells comprises administering the medicament to the patient in need thereof, and wherein the administration reduces the adhesion of red blood cells to endothelial cells expressing thrombospondin, VCAM-1, or laminin by at least 10% in the patient in need thereof after administration of the medicament.

21. The use according to claim 19 or claim 20, wherein the stored oxygen-depleted blood further comprises reduced carbon dioxide.

22. The use according to claim 19 or claim 20, wherein the patient in need thereof has sickle cell disease, inflammation, or both sickle cell disease and inflammation.

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