Methods for managing adverse events in patients with inflammation

Abstract

To provide a stored oxygen-reduced blood product for treating a patient in need thereof.SOLUTION: Provided is a stored oxygen-reduced blood product for use in a patient having inflammation and in need of treatment for increased C-Reactive Protein (CRP) levels, the stored oxygen-reduced blood product having an oxygen saturation of 20% or less during storage, the increased CRP level in the patient in need of the treatment being at least 3.0 milligrams per liter (mg / L), the patient in need of the treatment having or being at risk of developing a fever, and the increased CRP level in the patient in need of the treatment being reduced by at least 20% upon transfusion compared to the CRP level in an inflammatory patient receiving a conventionally stored non-oxygen-reduced blood product.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 768,667, filed November 16, 2018.

[0002] Government Rights This invention was made with United States Government support under R44HL132172 awarded by the National Heart, Lung, and Blood Institute. The Government has certain rights in this invention.

[0003] FIELD OF THEINVENTION The present disclosure relates to the treatment of inflammation, sickle cell disease, trauma, and hemorrhagic shock. [Background technology]

[0004] Sickle cell disease (SCD), a group of inherited red blood cell disorders, affects millions of people worldwide. It is estimated that approximately 100,000 Americans have SCD.

[0005] Sickle cell disease is divided into six types: sickle cell anemia (HbSS), sickle cell-hemoglobin C disease (HbSC), hemoglobin S-beta thalassemia (HbSβ0 and Hbβ+ thalassemia), hemoglobin-SD disease (HbSD), hemoglobin-SE disease (HbSE), and hemoglobin-SO Arab disease (HbSO). HbSS, the most severe form, 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. 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. bSC is the milder form of the disease. People with HbS beta thalassemia inherit one sickle gene and one gene for beta thalassemia. HbS beta thalassemia is further divided into beta thalassemia O and beta thalassemia +, with beta thalassemia O being the more severe form of the two. HbSD, HbSE, and HbSO are rare types of sickle cell disease. In these rare types, the sickle cell gene and one gene from an abnormal type of hemoglobin (D, E, or O) are inherited.

[0006] Some of the most common complications of SCD include vascular occlusion. Research has shown that sickle-shaped RBCs (SS-RBCs) cause vaso-occlusive episodes through adhesive interactions with the endothelium. The vaso-occlusive episodes cause pain of varying degrees depending on the person. This pain can begin suddenly and last for any period of time. In addition, patients are more likely to experience infections, including influenza, meningitis, and hepatitis. In addition, patients may experience hand-foot syndrome, eye disease, acute chest syndrome, and stroke. Furthermore, research has shown that sickle cell disease promotes inflammatory responses. See Non-Patent Document 1. Over a lifetime, SCD can harm the patient's brain, eyes, heart, lungs, liver, spleen, kidneys, skin, joints, and bones.

[0007] Treatments for sickle cell disease include blood transfusions, bone marrow transplants, stem cell transplants, and medications (eg, hydroxyurea) to reduce the number of painful episodes of crises.

[0008] In 2010, 5.1 million people died from injuries, more than the combined deaths from HIV, tuberculosis, and malaria (3.8 million). See Norton 2013, which is incorporated herein by reference in its entirety. Injuries include unintentional injuries (e.g., road traffic accidents, falls, and burns) and intentional injuries (e.g., self-injury, interpersonal violence, war, and conflict). See Norton 2013, which is incorporated herein by reference in its entirety. Deaths from injuries increased 24% worldwide between 1990 and 2010, and 23% in the United States between 2000 and 2010. See Norton 2013, which is incorporated herein by reference in its entirety. In addition, at least 20% of all trauma deaths are the result of survivable injuries and are therefore preventable with optimal care. Norton 2013, which is incorporated herein by reference in its entirety. This percentage of preventable deaths makes it imperative to develop treatments for avoidable complications that lead to death.

[0009] Penetrating wounds (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 state of inadequate oxygen delivery to tissues due to massive hemorrhage, causing oxygen debt, anaerobic metabolism, and elevated plasma lactate levels. Failure to reverse shock by restoring circulation and oxygen delivery can lead to permanent tissue damage, multiple organ failure, and death.

[0010] Clinical sequelae of hemorrhagic trauma and shock include death from exsanguination within hours of trauma, and death >24 hours after trauma and morbidity from massive transfusion, including multiple organ failure including lung, kidney, and liver due to acute traumatic coagulopathy or inflammation, and infection / sepsis due to transfusion-associated immunomodulation, both of which are increased by the poor quality of transfused blood products and the increased volume of transfused pRBCs.

[0011] One approach to treat hemorrhagic shock is the use of crystalloids for resuscitation. However, the use of crystalloids results in increased morbidity and mortality by causing trauma-induced coagulopathy. For at least this reason, early administration of blood components to reverse shock caused by hemorrhagic trauma is recommended. Packed red blood cells (pRBCs) are transfused into hemorrhagic trauma patients to restore lost blood volume, restore oxygen-carrying capacity in the patient, and restore oxidative metabolism from anaerobic metabolism in tissues. However, the use of pRBCs is not without risk of problems, including antigen mismatch, pathogen infection, circulatory overload, and degradation of pRBCs during ex vivo storage.

[0012] When conventionally stored, stored blood undergoes constant deterioration associated with various storage pathologies, including hemolysis, hemoglobin degradation, and reduced ATP and 2,3-DPG concentrations, among others. When transfused to a patient, the effects of constant deterioration during storage are manifested, for example, as reduced 24-hour in vivo recovery. The rapid decrease in hematocrit resulting from reduced 24-hour recovery can, when severe, result in delayed hemolytic transfusion reaction (DHTR). Other complications, such as systemic inflammatory response syndrome (SIRS), transfusion related acute lung injury (TRALI), and transfusion related immunomodulation (TRIM), are associated with transfusion of stored blood, but identification of the underlying causes remains unclear.

[0013] Even when transfused within the current 6-week time frame, stored RBCs tend to exhibit reduced quality (e.g., increased RBC fraction removed, decreased oxygen exchange capacity, reduced deformability) and increased toxicity, which often manifests as clinical sequelae of transfusion therapy. A large and growing number of articles in the literature support this view. See Fischer et al., 2004, 2005, 2006, 2007, 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 2020, 2021 (incorporated herein by reference in their entirety). Extensive in vitro studies clearly demonstrate the degradation of RBCs during traditional storage (storage lesion). A number of emerging metabolomic studies demonstrate the progression of storage lesion at the molecular level. See, e.g., 22, 23, 24, 25, 26, 27, 28, which are incorporated by reference in their entireties. There is a need to reduce or prevent this degradation to increase transfusion efficacy (more O2 delivery to peripheral tissues immediately after transfusion) and reduce mortality from inflammation, sickle cell disease, or hemorrhagic trauma.

[0014] Oxidative damage causes many RBC storage lesions and their downstream consequences in conventionally stored blood. Therefore, methods are needed to reduce the degree of oxidative stress in order to reduce RBC storage lesions. Several approaches have been developed with the aim of minimizing storage lesions and improving transfusion outcomes. These include additive solutions (e.g., U.S. Patent No. 5,333,439 to Hamasaki et al., U.S. Patent No. 5,333,439 to Sasakawa et al., U.S. Patent No. 5,333,439 to Hess et al.), cryopreservation (see U.S. Patent No. 5,333,439 to Serebrennikov, U.S. Patent No. 5,333,439 to Serebrennikov, and U.S. Patent No. 5,333,439 to Serebrennikov, which are incorporated herein by reference in their entirety).

[0015] One approach that has proven successful in improving the quality of blood and expanding its usefulness is through storage under oxygen-depleted and anaerobic conditions. Among the benefits of storing blood under oxygen-depleted conditions are improved levels of ATP and 2,3-DPG, and reduced hemolysis. U.S. Patent No. 5,393,433 to Bitensky et al., U.S. Patent No. 5,393,466 to Bitensky et al., and U.S. Patent No. 5,393,466 to Bitensky et al., which are incorporated herein by reference in their entirety, are directed to the storage of red blood cells under oxygen-depleted conditions. U.S. Patent No. 5,393,433 to Bitensky et al., which are incorporated herein by reference in their entirety, are directed to blood storage additive solutions. U.S. Patent No. 5,393,433 to Bitensky et al. ('396 patent), which is incorporated herein by reference in its entirety, discloses an anaerobic storage bag for blood storage that includes an oxygen-impermeable outer layer, an oxygen-permeable red blood cell (RBC)-compatible inner layer, and has an oxygen scrubber disposed between the inner and outer layers.

[0016] Storage of blood under oxygen-depleted conditions may also result in reduced particulate levels, reduced loss of deformability, reduced lipid and protein oxidation, and higher post-transfusion survival rates compared to blood stored under conventional conditions. See Non-Patent Document 31 and Non-Patent Document 32, which are incorporated herein by reference in their entirety. Anaerobically stored RBCs further provide higher 24-hour in vivo recovery rates after autotransfusion, higher 2,3-DPG and ATP levels, lower hemolysis, and beneficial remodeling of metabolic pathways. See Non-Patent Document 33 and Non-Patent Document 34, which are incorporated herein by reference in their entirety.

[0017] In this 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. Additionally, we demonstrate that OR and OCR red blood cells reduced hemolysis in the presence of plasma from SCD patients compared to conventionally stored blood.

[0018] In this disclosure, we demonstrate that OR or OCR blood from rats provides improved ATP and 2,3-DPG during storage compared to conventionally stored blood, as previously demonstrated using human blood. Thus, OR or OCR rat RBCs are expected to have similar reduced particulates, improved deformability, reduced lipid and protein oxidation, and higher post-transfusion survival.

[0019] Here, we demonstrate for the first time that OR and OCR blood in rats provide a surprising improvement in clinical outcomes when transfused to treat hemorrhagic trauma. Using a rat hemorrhagic shock resuscitation model, we show that OR or OCR RBCs provide reduced organ damage than conventionally stored blood. In addition, OR or OCR RBCs provide recovery from shock conditions using a smaller amount of pRBCs. Finally, OR or OCR RBCs stabilize hemodynamics more rapidly when transfused to treat hemorrhagic shock compared to conventionally stored pRBCs of the same storage period.

[0020] OR and OCR RBCs provide an improved method for the treatment of trauma resulting in blood loss to reduce mortality and morbidity over 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 provide a reduction in the volume of blood required to restore and stabilize hemodynamic function. Thus, OR and OCR RBCs may provide a reduction in the volume of RBCs required for transfusion therapy when treating hemorrhagic trauma. The quality improvements of OR and OCR, in addition to the previously demonstrated improvements to the ability of stored RBCs to deliver oxygen, also provide an unexpected reduction in trauma-related organ damage, morbidity, and mortality. [Prior art documents] [Patent documents]

[0021] [Patent Document 1] U.S. Patent No. 4,769,318 [Patent Document 2] U.S. Patent No. 4,880,786 [Patent Document 3] U.S. Patent No. 6,447,987 [Patent Document 4] U.S. Patent No. 6,413,713 [Patent Document 5] U.S. Patent No. 5,624,794 [Patent Document 6] U.S. Patent No. 6,162,396 [Patent Document 7] U.S. Patent No. 5,476,764 [Patent Document 8] U.S. Patent No. 5,789,151 [Non-patent literature]

[0022] [Non-Patent Document 1] Plat, O., "Sickle cell anemia as an inflammatory disease," J. Clin. Invest., 106(3):337-338 (2000) [Non-Patent Document 2] Norton et al., "Global Health Injuries," The NEJM, 368:1723-30 (2013) [Non-Patent Document 3] Fox et al., “Earlier Endpoints are Required for Hemorrhagic Shock Trials Among Severely Injured Patients.” Shock, 47:567-73 (2017) [Non-Patent Document 4] Zimring, "Established and theoretical factors to consider in assessing the red cell storage lesion," Blood, 125:2185-90 (2015) [Non-Patent Document 5] 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) [Non-Patent Document 6] Weinberg et al., "Red blood cell age and potentiation of transfusion-related pathology in trauma patients," Transfusion, 51:867-73 (2011) [Non-Patent Document 7] Spinella et al., "Does the storage duration of blood products affect outcomes in critically ill patients?" 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[0023] The present disclosure provides and includes a method of treating a patient in need thereof with stored oxygen-depleted blood having an oxygen saturation of 20% or less during storage, wherein the patient in need thereof has inflammation.

[0024] The present disclosure provides and includes a method of improving blood transfusion in a sickle cell patient in need thereof, comprising providing stored oxygen-reduced blood to a patient with sickle cell disease, the oxygen-reduced blood having an oxygen saturation of 20% or less during storage.

[0025] The present disclosure provides and includes a method of reducing the number of vaso-occlusive episodes in a patient in need thereof comprising providing stored oxygen-reduced blood to the patient in need thereof, the oxygen-reduced blood having an oxygen saturation of 20% or less during storage, and reducing the vaso-occlusive episodes comprises reducing adhesion of red blood cells to endothelial cells.

[0026] The present disclosure provides and includes a method of reducing adhesion of red blood cells to endothelial cells in a patient in need thereof comprising providing stored oxygen-reduced blood to a patient with sickle cell disease, the oxygen-reduced blood having an oxygen saturation of 20% or less during storage.

[0027] The present disclosure is provided with reference to the accompanying drawings. [Brief description of the drawings]

[0028] [Figure 1] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, comparing 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). [Diagram 2] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, comparing 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). [Diagram 3] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, illustrating a comparison of the percent recovery of control, sham, OR-RBCs, and OCR-RBCs transfused into animals. [Figure 4A]1 is a graph showing results of an exemplary embodiment according to the present disclosure, showing a comparison of percent hematocrit in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. BL (baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 4B] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, showing a comparison of percent hematocrit in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. BL (Baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 5A] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of mean arterial pressure (MAP) in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. BL (Baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 5B] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of mean arterial pressure (MAP) in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. BL (Baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 6A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the percent blood volume donated to animals during resuscitation at 10, 20, 30, 45, and 60 minutes, comparing control, OR-RBCs, and OCR-RBCs stored for 1 week. [Figure 6B]1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the percent blood volume provided to animals during resuscitation at 10, 20, 30, 45, and 60 minutes, comparing control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 7A] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of lactate in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. BL (Baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 7B] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of lactate in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. BL (Baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 8A] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of glucose in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. BL (Baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 8B] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of glucose in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. BL (Baseline) identifies animals not in shock conditions. Shock identifies animals in hemorrhagic shock. Early R identifies a 10 minute resuscitation period. Late R identifies a 60 minute resuscitation period. [Figure 9A]1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of AST in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 9B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of AST in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 10A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of ALT in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 10B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of ALT in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 11A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of serum creatinine levels in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 11B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of serum creatinine levels in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 12A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of blood urea nitrogen (BUN) amounts in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 12B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of blood urea nitrogen (BUN) amounts in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 13A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of CXCL1 in the liver of animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 13B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of CXCL1 in the liver of animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 14A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of CXCL1 in the spleens of animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 14B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of CXCL1 in the spleens of animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 15A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of CXCL1 in the lungs of animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 15B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of CXCL1 in the lungs of animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 16A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of urinary neutrophil gelatinase-associated lipocalin (u-NGAL) in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 16B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of urinary neutrophil gelatinase-associated lipocalin (u-NGAL) in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 17A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the percentage of CD45+ neutrophils in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 17B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the percentage of CD45+ neutrophils in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 18A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of IL-6 in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for one week. [Figure 18B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of the amount of IL-6 in animals resuscitated with control, OR-RBCs, and OCR-RBCs stored for 3 weeks. [Figure 19A] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of red blood cell adhesion to thrombospondin. [Figure 19B] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of red blood cell adhesion to thrombospondin. [Figure 19C] 1 is a graph showing results of an exemplary embodiment according to the present disclosure, providing a comparison of red blood cell adhesion to thrombospondin. [Figure 20A] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of red blood cell adhesion to thrombospondin between conventionally stored red blood cells or red blood cells stored with reduced oxygen. [Figure 20B] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of red blood cell adhesion to thrombospondin between conventionally stored red blood cells or red blood cells stored with reduced oxygen. [Figure 20C] 1 is a graph showing the results of an exemplary embodiment according to the present disclosure, providing a comparison of red blood cell adhesion to thrombospondin between conventionally stored red blood cells or red blood cells stored with reduced oxygen. [Figure 21]FIG. 1 shows results of an exemplary embodiment according to the present disclosure, providing a comparison of hemolysis in conventionally stored red blood cells, reduced oxygen stored red blood cells, and reduced oxygen and carbon dioxide stored red blood cells incubated with healthy sickle cell plasma. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] The examples described herein illustrate some embodiments of the present disclosure, but should not be construed as limiting the scope of the disclosure in any way.

[0030] The disclosed method provides and includes providing a bleeding patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. Additionally, the method provides for providing a bleeding patient with oxygen-reduced stored blood having an oxygen saturation of 15-20% before and during storage. Additionally, the method provides for providing a bleeding patient with oxygen-reduced stored blood having an oxygen saturation of 10-15% before and during storage. Additionally, the method provides for providing a bleeding patient with oxygen-reduced stored blood having an oxygen saturation of 5-10% before and during storage. Additionally, the method provides for providing a bleeding patient with oxygen-reduced stored blood having an oxygen saturation of 3-5% before and during storage.

[0031] Additionally, the method provides for providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage for transfusion to a person with hemorrhagic shock. Additionally, the method provides for providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage for transfusion to a person with hemorrhagic trauma. Also included is a method including transfusing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient at increased risk of trauma from 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 further 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 further includes providing oxygen-reduced stored blood having an initial oxygen saturation of 3% or less.

[0032] The disclosed method provides and includes providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 20% or less before and during storage for a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. Additionally, the method provides for providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 15% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. Additionally, the method provides for providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 10% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method further provides for providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 5% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method further provides for providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 3% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. Additionally, the method provides for providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 3-5% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. Additionally, the method provides for providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 5-10% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. Additionally, the method provides for providing oxygen-reduced stored blood for the treatment of trauma having an oxygen saturation of 10-15% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks.Additionally, the method provides for providing reduced oxygen stored blood for treatment of trauma having an oxygen saturation of 15-20% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks.

[0033] The disclosed method provides and includes providing a trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage.In one embodiment, the trauma patient suffers from head trauma, penetrating wound, blunt trauma, fall injury, or automobile accident injury.In another embodiment, the trauma patient is a bleeding trauma patient.In yet another embodiment, the trauma patient is a bleeding trauma patient from surgery, penetrating wound, blunt trauma, fall injury, or automobile accident injury.

[0034] In one embodiment of the present disclosure, the trauma patient or hemorrhagic trauma patient is a subject who needs OR and OCR stored blood. In an embodiment of the present disclosure, the trauma patient is a hemorrhagic trauma patient who needs one or more units of blood as transfusion therapy. In an embodiment of the present disclosure, the trauma patient is a hemorrhagic trauma patient who needs two or more units of blood as transfusion therapy. In an embodiment of the present disclosure, the trauma patient is a hemorrhagic trauma patient who needs three or more units of blood as transfusion therapy.

[0035] In one embodiment of the present disclosure, the trauma patient is a patient in hemorrhagic shock. In one embodiment, the trauma patient is in hemorrhagic shock due to head trauma, penetrating wound, blunt trauma, injury from a fall, or injury from a motor vehicle accident. In an embodiment of the present disclosure, the hemorrhagic trauma patient is a patient with class I hemorrhage. In another embodiment, the hemorrhagic trauma patient is a patient with class II hemorrhage. In another embodiment, the hemorrhagic trauma patient is a patient with class III hemorrhage. In another embodiment, the hemorrhagic trauma patient is a patient with class IV hemorrhage. In one embodiment of the present disclosure, the hemorrhagic trauma patient loses up to 15% of the blood volume. In another embodiment, the hemorrhagic trauma patient loses 15-30% of the blood volume. In another embodiment, the hemorrhagic trauma patient loses 30-40% of the blood volume. Additionally, the hemorrhagic trauma patient loses more than 40% of the blood volume.

[0036] The present disclosure provides and includes a patient in need of transfusion therapy with OR or OCR RBCs, who shows one or more symptoms 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 embodiment of the present disclosure, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with decreased mean arterial pressure. The present disclosure provides and includes a patient in need of transfusion therapy with OR or OCR RBCs, who shows increased aspartate aminotransferase (AST) and increased alanine aminotransferase (ALT). The present disclosure provides and includes a patient in need of transfusion therapy with OR or OCR RBCs, who shows decreased mean arterial pressure and increased lactate. The present disclosure provides and includes patients in need of transfusion therapy with OR or OCR RBCs exhibiting increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), and increased blood urea nitrogen. The present disclosure provides and includes patients in need of transfusion therapy with OR or OCR RBCs exhibiting increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), increased serum creatinine, and increased blood urea nitrogen. The present disclosure provides and includes patients in need of transfusion therapy with OR or OCR RBCs exhibiting increased lactate and increased glucose. The present disclosure provides and includes patients in need of transfusion therapy with OR or OCR RBCs exhibiting increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL), increased serum creatinine, and increased blood urea nitrogen.

[0037] In another embodiment, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with a decreased hematocrit. In another embodiment, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with an increased lactate. In yet another embodiment, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with an increased glucose. In a further embodiment, the hemorrhagic trauma patient has an increased aspartate aminotransferase (AST). In another embodiment, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with an increased alanine aminotransferase (ALT). In another embodiment, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with an increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL). In another embodiment, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with an increased serum creatinine. In another embodiment, the patient in need of transfusion therapy with OR or OCR RBCs is a hemorrhagic trauma patient with elevated blood urea nitrogen.

[0038] In one embodiment of the disclosure, OR and OCR stored blood for use in transfusion therapy of a patient in need thereof has an initial oxygen saturation of 20% or less. In another embodiment, OR and OCR stored blood has an initial oxygen saturation of 10% or less. In another embodiment, OR and OCR stored blood has an initial oxygen saturation of 5% or less. In another embodiment, OR and OCR stored blood has an initial oxygen saturation of 3% or less.

[0039] In one embodiment of the disclosure, OCR-stored blood for use in transfusion therapy of a patient in need thereof has an initial pCO2 (at 37°C) of 10-40 mmHg. In another embodiment, the OCR-stored blood has an initial pCO2 of 10-30 mmHg. In another embodiment, the OCR-stored blood has an initial pCO2 of 10-20 mmHg. In another embodiment, the OCR-stored blood has an initial pCO2 of 10-15 mmHg. In yet another embodiment, the OCR-stored blood has an initial pCO2 of less than 10 mmHg.

[0040] In one embodiment of the disclosure, OR and OCR stored blood for use in transfusion therapy of a patient in need has an initial oxygen saturation of 20% or less and is stored for less than 2 days. In one embodiment, OR and OCR stored blood has an initial oxygen saturation of 20% or less and is stored for less than 7 days. In another embodiment, OR and OCR stored blood with an initial oxygen saturation of 20% or less is stored for less than 14 days. In another embodiment, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 21 days. In another embodiment, oxygen-reduced stored blood for use in transfusion therapy of a patient in need has an initial oxygen saturation of 20% or less and is stored for less than 28 days. In another embodiment, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 35 days. In another embodiment, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 42 days. In another embodiment, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 45 days. In one embodiment of the disclosure, OR and OCR stored blood has an oxygen saturation of 20% or less during storage.

[0041] Suitable blood for use in transfusion therapy of a patient in need thereof includes oxygen-reduced stored blood with an anticoagulant. In one embodiment of the present disclosure, oxygen-reduced red blood cells are stored for up to three weeks to generate oxygen-reduced stored blood. In another embodiment, the oxygen-reduced stored blood typically further comprises an additive solution. Suitable additive solutions according to the present disclosure include AS-1, AS-3 (Nutricel®), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, OFAS3, and combinations thereof. In one embodiment, the additive solution is added during component separation. In one embodiment, the additive solution is AS-1. In another embodiment, the additive solution is AS-3. In another embodiment, the additive solution is SAGM.

[0042] The disclosed method provides and includes increasing the mean arterial pressure (MAP) of a hemorrhagic trauma patient in need of transfusion therapy comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the MAP is increased by 20-60%. In another embodiment, the MAP is increased by 30-60%. In another embodiment, the MAP is increased by 30-50% in a trauma patient receiving transfusion therapy with OR or OCR blood. In yet another embodiment, the MAP is increased by 30-60%. In a further embodiment, the MAP is increased by 30-40% in a trauma patient receiving transfusion therapy with OR or OCR blood. In one embodiment, the MAP is increased by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90% over the MAP of a patient transfused with conventionally stored blood.

[0043] In one embodiment of the disclosure, the mean arterial pressure of a patient receiving transfusion treatment with OR or OCR blood is increased by at least 1.5-fold. In another embodiment, the mean arterial pressure of a trauma patient receiving transfusion treatment with OR or OCR blood is increased by at least 2-fold. In a further embodiment, the mean arterial pressure of a patient receiving transfusion treatment with OR or OCR blood is increased by 1-2-fold. In one embodiment of the disclosure, the mean arterial pressure of a trauma patient receiving transfusion treatment 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 embodiment, the mean arterial pressure of a trauma patient receiving transfusion treatment with OR or OCR blood is increased by 20-50 mmHg. In a further embodiment, the mean arterial pressure of a patient receiving transfusion treatment with OR or OCR blood is increased by 30-50 mmHg.

[0044] The disclosed method provides and includes increasing the mean arterial pressure of a trauma patient in need of transfusion therapy to 70-110 mmHg, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In another embodiment, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased to at least 70 mmHg. In another embodiment, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased to at least 80 mmHg. In yet another embodiment, the mean arterial pressure is increased to at least 90 mmHg. In a further embodiment, the mean arterial pressure is increased to at least 100 mmHg. In one embodiment of the disclosure, the mean arterial pressure in a subject in need remains at 70-110 mmHg for at least 1 hour after transfusion. In another embodiment, the mean arterial pressure remains at 70-110 mmHg for at least 2 hours after transfusion. In yet another embodiment, the mean arterial pressure remains at 70-105 mmHg for at least 3 hours after transfusion. In another embodiment, the mean arterial pressure remains between 70 and 110 mmHg for at least 4 hours after the transfusion.In another embodiment, the mean arterial pressure remains between 70 and 110 mmHg for at least 5 hours after the transfusion.

[0045] The disclosed method provides and includes increasing mean arterial pressure in a trauma patient in need of transfusion therapy at a faster rate than the mean arterial pressure of a patient transfused with conventionally stored blood, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the mean arterial pressure of a patient transfused with OR or OCR blood recovers to within normal physiological parameters in half the time compared to conventionally stored blood.

[0046] The disclosed method provides and includes reducing the amount of stored blood required for transfusion in a hemorrhagic trauma patient in need of transfusion therapy, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the amount of OR-stored blood required for transfusion is 10-90% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is 10-30% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is 20-50% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is 20-80% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is 30-80% less than the amount of conventionally stored blood required. In yet another embodiment, the amount of OR-stored blood required for transfusion is 40-85% less than the amount of conventionally stored blood required. In a further embodiment, the amount of OR stored blood required for transfusion is 50-90% less than the amount of conventionally stored blood required.

[0047] The disclosed method 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%, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the amount of OR-stored blood required for transfusion is at least 20% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is at least 30% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is at least 40% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is at least 50% less than the amount of conventionally stored blood required. In yet another embodiment, the amount of OR-stored blood required for transfusion is at least 60% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion is at least 70% less than the amount of conventionally stored blood required. In further embodiments, the amount of OR stored blood required for transfusion is about 10-20%, about 20-30%, about 30-40%, about 40-50%, about 50-60%, about 60-70%, about 70-80%, about 80-90%, or about 90-95% less than the amount of conventionally stored blood required. In other embodiments, the amount of OR stored blood required for transfusion of a patient in need of transfusion therapy is 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% less than the amount of conventionally stored blood required.

[0048] Lactate clearance is a biomarker for resuscitation from hemorrhagic shock. 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," Journal of Trauma and Acute Care Surgery, 76:894-901 (2014); See, "A meta-analysis," Critical care medicine, 42:2118-25 (2014) ("Chang 2014"), incorporated herein by reference in its entirety. The clinical value of lactate clearance is useful in predicting the prognosis of patients with septic shock and critically ill patients without overt circulatory shock. Elevated lactate is an indicator of adverse clinical prognosis, and its rapid clearance is universally associated with improved prognosis in heterogeneous ICU or ED patient populations. See, Chang 2014. The reduced lactate levels in animals resuscitated with OR-RBCs compared with conventional RBCs supports the notion that resuscitation with OR RBCs may significantly improve patient clinical prognosis.Please refer to Figures 7A and 7B.

[0049] The disclosed methods provide and include reducing lactate levels in a patient in need of transfusion therapy comprising providing the patient with oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less prior to and during storage. In one embodiment, lactate levels are reduced by 10-90%. In one embodiment, transfusion with OR-stored blood reduces lactate levels in a patient in need of transfusion therapy by 10-50%. In another embodiment, transfusion with OR-stored blood reduces lactate levels in a patient in need of transfusion therapy by 20-40%. In another embodiment, transfusion with OR-stored blood reduces lactate levels in a patient in need of transfusion therapy by 50-90%. In yet another embodiment, transfusion with OR-stored blood reduces lactate levels in a patient in need of transfusion therapy by 60-90%. In another embodiment, transfusion with OR-stored blood reduces lactate levels in a patient in need of transfusion therapy by 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90%. In another embodiment, transfusion with OR-stored blood reduces lactate levels in a trauma patient in need of transfusion therapy by at least 10%. In another embodiment, transfusion with OR-stored blood reduces lactate levels in a patient in need of transfusion therapy by at least 20%. In further embodiments, transfusion with OR-stored blood reduces lactate levels in a patient in need of 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%.

[0050] The disclosed method provides and includes reducing elevated lactate levels in a patient in need of transfusion therapy to about 0.5 to about 2.5 mmol / L, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the lactate level in a patient in need of transfusion therapy is reduced to about 0.9 to about 2 mmol / L. In one embodiment, the lactate level in a patient in need of transfusion therapy is reduced to about 0.9 to about 1.7 mmol / L. In another embodiment, the lactate level in a patient in need of transfusion therapy is reduced to about 1.4 to about 2.4 mmol / L. In another embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to about 1.7 to about 2.5 mmol / L. In yet another embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to less than about 2.5 mmol / L. In a further embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to less than about 2.0 mmol / L. In another embodiment, a trauma patient in need of transfusion therapy has lactate levels reduced to less than about 1.5 mmol / L. In another embodiment, a trauma patient in need of transfusion therapy has lactate levels reduced to less than about 1.0 mmol / L. In yet another embodiment, a trauma patient in need of transfusion therapy has lactate levels reduced to between about 0.5 and about 1.0 mmol / L.

[0051] The disclosed method provides and includes reducing elevated lactate levels in a trauma patient in need of transfusion therapy to between 0.5-2.5 mmol / L, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to between 0.9-2 mmol / L. In one embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to between 0.9-1.7 mmol / L. In another embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to between 1.4-2.4 mmol / L. In another embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to between 1.7-2.5 mmol / L. In another embodiment, the lactate level in a trauma patient in need of transfusion therapy is reduced to between 0.5-1 mmol / L.

[0052] The disclosed method provides and includes reducing elevated lactate levels in a hemorrhagic trauma patient in need of transfusion therapy to less than 4 mmol / L, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the lactate level of a trauma patient in need of transfusion therapy is reduced to less than 3 mmol / L. In yet another embodiment, the lactate level of a trauma patient in need of transfusion therapy is reduced to less than 2.5 mmol / L. In another embodiment, the patient's lactate level is reduced to less than 2.3 mmol / L. In another embodiment, the lactate level of a trauma patient in need of transfusion therapy is reduced to less than 2 mmol / L. In another embodiment, the lactate level of a trauma patient in need of transfusion therapy is reduced to less than 2 mmol / L. In another embodiment, the lactate level of a trauma patient in need of transfusion therapy is reduced to less than 1.5 mmol / L. In another embodiment, the lactate level of a trauma patient in need of transfusion therapy is reduced to less than 1 mmol / L.

[0053] Blood glucose levels are also known to be predictive of outcomes in some disease patterns, especially in trauma patients. Trauma patients are more susceptible to poor outcomes due to hyperglycemia than other critically ill patients. 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 between early hyperglycemia and trauma patients have tested for early hyperglycemia at three possible cutoffs: 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 were associated with significantly higher infection and mortality in trauma patients, independent of injury characteristics. This was not true for cutoffs of ≧110 mg / dL or ≧150 mg / dL. The reduced glucose levels in animals resuscitated with OR-RBCs and OCR-RBCs compared to conventional RBCs supports the notion that resuscitation with OR-RBCs may significantly improve patient clinical outcomes. See Figures 8A and 8B.

[0054] The disclosed methods provide and include reducing glucose in a trauma patient in need of transfusion therapy, comprising providing to the trauma patient oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, glucose is reduced by 10-90% compared to transfusion of blood stored under conventional conditions. In one embodiment, transfusion with OR-stored blood reduces glucose by 10-50% compared to transfusion of blood stored under conventional conditions. In another embodiment, transfusion with OR-stored blood reduces glucose by 20-40% compared to transfusion of blood stored under conventional conditions. In another embodiment, transfusion with OR-stored blood reduces glucose by 50-90% compared to transfusion of blood stored under conventional conditions. In yet another embodiment, transfusion with OR-stored blood reduces glucose by 60-90%. In another embodiment, transfusion with OR stored blood reduces glucose by 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90% compared to transfusion of blood stored under conventional conditions. In another embodiment, transfusion with OR stored blood reduces glucose by at least 10% compared to transfusion of blood stored under conventional conditions. In another embodiment, transfusion with OR stored blood reduces glucose by at least 20%. In further embodiments, transfusion with OR stored blood 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%.

[0055] The disclosed method provides and includes reducing the glucose level of a trauma patient in need of transfusion therapy to about 70 to about 120 mg / dL, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is about 70 to about 110 mg / dL. In another embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is about 70 to about 100 mg / dL. In another embodiment, the trauma patient's glucose after transfusion therapy with OR or OCR blood is about 90 to about 120 mg / dL. In another embodiment, the trauma patient's glucose after transfusion therapy with OR or OCR blood is about 90 to about 100 mg / dL.

[0056] The disclosed methods provide and include reducing glucose levels of a trauma patient in need of transfusion therapy to between 70-120 mg / dL including providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is between 70-110 mg / dL. In another embodiment, the patient's glucose is between 70-100 mg / dL. In another embodiment, the patient's glucose is between 90-120 mg / dL. In another embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is between 90-100 mg / dL.

[0057] The disclosed method provides and includes reducing glucose levels of a trauma patient in need of transfusion therapy to less than 120 mg / dL, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In a further embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is less than 110 mmol / L. In yet another embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is less than 100 mg / dL. In another embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is less than 200 mg / dL. In another embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is less than 90 mg / dL. In another embodiment, the patient's glucose after transfusion therapy with OR or OCR blood is less than 80 mg / dL.

[0058] In one embodiment of the present disclosure, the patient is at increased risk of complications from transfusion therapy due to a pre-existing or underlying condition.In one embodiment, the patient has a pre-existing or underlying 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, chronic pulmonary obstructive disease such as smoke inhalation, systemic inflammation caused by infection, coagulation disorder, and autoimmune disease.In another embodiment, the patient has one or more pre-existing or underlying 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, systemic inflammation caused by infection, chronic pulmonary obstructive disease such as coagulation disorder, and autoimmune disease.In another embodiment, the patient has two or more pre-existing or underlying 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, systemic inflammation caused by infection, chronic pulmonary obstructive disease such as coagulation disorder, and autoimmune disease. In another embodiment, the patient has three or more pre-existing or underlying conditions selected from the group consisting of diabetes, ischemic heart disease, systemic inflammatory syndrome resulting from trauma or infection, multiple organ failure resulting from trauma or infection, chronic pulmonary obstructive disease such as smoke inhalation, systemic inflammation due to infection, coagulation disorders, and autoimmune diseases.

[0059] During hemorrhagic shock, patients experience adverse events, including liver damage or failure, kidney damage or failure, lung damage or failure, or combinations thereof.The present disclosure provides and includes patients in need of transfusion therapy with OR or OCR RBCs, who show one or more adverse events selected from the group consisting of liver damage or failure, kidney damage or failure, or lung damage or failure.The present disclosure provides and includes patients in need, who show two or more adverse events selected from the group consisting of liver damage or failure, kidney damage or failure, or lung damage or failure.

[0060] The disclosed method provides and includes 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 a trauma patient in need of transfusion therapy. In one embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 5%. In another embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 10%. In another embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 20%. In another embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 30%. In another embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 40%. In another embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 50%. In another embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 60%. In another embodiment, adverse events are reduced by at least 70%. In another embodiment, adverse events following transfusion therapy with OR or OCR blood are reduced by at least 80%. In another embodiment, adverse events are reduced by at least 90%. In further embodiments, adverse events following transfusion therapy with OR or OCR blood are reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95%. In one embodiment, the adverse event following transfusion therapy with OR or OCR blood is liver injury or damage. In another embodiment, the adverse event is lung injury or damage. In yet another embodiment, the adverse event is kidney injury or damage. In further embodiments, the adverse event is liver injury, lung injury, kidney injury, or a combination thereof.

[0061] 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 disclosed method provides and includes reducing elevated levels of liver enzymes in trauma patients, including providing trauma patients with oxygen-reduced preserved blood having an oxygen saturation of 20% or less before and during storage.

[0062] The disclosed method provides and includes reducing AST levels in trauma patients in need of transfusion therapy, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the AST level is reduced by at least 5% compared to the AST level of a patient transfused with conventionally stored blood. In another embodiment, the AST level is reduced by at least 10% compared to the AST level of a patient transfused with conventionally stored blood. In another embodiment, the AST level is reduced by at least 20% compared to the AST level of a patient transfused with conventionally stored blood. In another embodiment, the AST level is reduced by at least 30% compared to the AST level of a patient transfused with conventionally stored blood. In another embodiment, the AST level is reduced by at least 40%. In another embodiment, the AST level is reduced by at least 50% compared to the AST level of a patient transfused with conventionally stored blood. In another embodiment, the AST level is reduced by at least 60%. In another embodiment, the AST level is reduced by at least 70% compared to the AST level of a patient transfused with conventionally stored blood. In yet another embodiment, AST levels are reduced by at least 80%. In a further embodiment, AST levels are reduced by at least 90% compared to AST levels in patients transfused with conventionally stored blood. In further embodiments, AST levels are reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to AST levels in patients transfused with conventionally stored blood.

[0063] The disclosed method provides and includes reducing AST levels in a trauma patient in need of transfusion therapy by 1.5-10 fold, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the AST levels are reduced by 2-3 fold compared to the AST levels of a patient transfused with conventionally stored blood. In another embodiment, the AST levels are reduced by 3-4 fold. In another embodiment, the AST levels are reduced by 4-10 fold. In another embodiment, the AST levels are reduced by 6-9 fold compared to the AST levels of a patient transfused with conventionally stored blood. In a further embodiment, the AST levels are reduced by 2-5 fold. In another embodiment, the AST levels are reduced by 10-50 fold compared to the AST levels of a patient transfused with conventionally stored blood.

[0064] The disclosed method provides and includes reducing AST levels in trauma patients in need of transfusion therapy by at least 1.5-fold, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the AST levels are reduced by at least 2-fold compared to the AST levels of patients transfused with conventionally stored blood. In another embodiment, the AST levels are reduced by at least 3-fold compared to the AST levels of patients transfused with conventionally stored blood. In another embodiment, the AST levels are reduced by at least 4-fold compared to the AST levels of patients transfused with conventionally stored blood. In another embodiment, the AST levels are reduced by at least 5-fold compared to the AST levels of patients transfused with conventionally stored blood. In a further embodiment, the AST levels are reduced by at least 6-fold. In another embodiment, the AST levels are reduced by at least 7-fold compared to the AST levels of patients transfused with conventionally stored blood. In another embodiment, the AST levels are reduced by at least 8-fold. In another embodiment, the AST levels are reduced by at least 9-fold compared to the AST levels of patients transfused with conventionally stored blood. In another embodiment, the AST levels are reduced by at least 10-fold compared to the AST levels of patients transfused with conventionally stored blood, hi a further embodiment, the AST levels are reduced by at least 50-fold compared to the AST levels of patients transfused with conventionally stored blood.

[0065] The disclosed method provides and includes reducing ALT levels in trauma patients requiring transfusion therapy, comprising providing oxygen-reduced stored blood with an oxygen saturation of 20% or less before and during storage to the trauma patient. In one embodiment, the ALT level is reduced by at least 5% compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 10% compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 20% compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 30% compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 40% compared to the AS ALT T level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 50% compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 60% compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 70% compared to the ALT level of a patient transfused with conventionally stored blood. In yet another embodiment, the ALT level is reduced by at least 80% compared to the ALT level of a patient transfused with conventionally stored blood. In a further embodiment, the ALT level is reduced by at least 90%. In a further embodiment, the ALT level is reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the ALT level of a patient transfused with conventionally stored blood.

[0066] The disclosed method provides and includes reducing ALT levels by 1.5-10 fold in trauma patients in need of transfusion therapy, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the ALT levels are reduced by 2-3 fold compared to ALT levels in patients transfused with conventionally stored blood. In another embodiment, the ALT levels are reduced by 3-4 fold compared to ALT levels in patients transfused with conventionally stored blood. In another embodiment, the ALT levels are reduced by 4-10 fold compared to ALT levels in patients transfused with conventionally stored blood. In another embodiment, the ALT levels are reduced by 6-9 fold compared to ALT levels in patients transfused with conventionally stored blood. In a further embodiment, the ALT levels are reduced by 2-5 fold. In another embodiment, the ALT levels are reduced by 10-50 fold compared to ALT levels in patients transfused with conventionally stored blood.

[0067] The disclosed method provides and includes reducing ALT levels in trauma patients requiring transfusion therapy by at least 1.5-fold, comprising providing the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the ALT levels are reduced by at least 2-fold compared to the ALT levels of patients transfused with conventionally stored blood. In another embodiment, the ALT levels are reduced by at least 3-fold compared to the ALT levels of patients transfused with conventionally stored blood. In another embodiment, the ALT levels are reduced by at least 4-fold compared to the ALT levels of patients transfused with conventionally stored blood. In another embodiment, the ALT levels are reduced by at least 5-fold compared to the ALT levels of patients transfused with conventionally stored blood. In a further embodiment, the ALT levels are reduced by at least 6-fold compared to the ALT levels of patients transfused with conventionally stored blood. In another embodiment, the ALT levels are reduced by at least 7-fold compared to the ALT levels of patients transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 8-fold compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 9-fold compared to the ALT level of a patient transfused with conventionally stored blood. In another embodiment, the ALT level is reduced by at least 10-fold compared to the ALT level of a patient transfused with conventionally stored blood. In a further embodiment, the ALT level is reduced by at least 50-fold compared to the ALT level of a patient transfused with conventionally stored blood.

[0068] Markers of renal 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 analysis reported in over 150 separate studies conducted in AKI models from several species ranging from rodents to humans has consistently revealed that the NGAL gene is one of the most dramatically upregulated genes in the kidney immediately following ischemic or nephrotoxic insults. 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 may vary depending on age, race, and body size, but elevated creatinine levels indicate kidney injury. Creatinine levels above 1.2 in women and 1.4 in men may be early signs of kidney injury. Increased blood urea nitrogen (BUN) is seen in association with renal disease or failure, as well as congestive heart failure, shock, and gastrointestinal bleeding.When BUN levels are higher than 100 mg / dL, it indicates severe kidney damage. A low 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), which are incorporated by reference in their entireties. As provided by the present disclosure, the reduction in u-NGAL (FIGS. 16A and 16B), serum creatinine (FIGS. 11A and 11B), and BUN (FIGS. 12A and 12B) levels in animals resuscitated with OR-RBCs and OCR-RBCs compared to conventional RBCs indicates that resuscitation with OR-RBCs may significantly improve patient clinical outcomes.

[0069] The disclosed method provides and includes reducing urinary neutrophil gelatinase-associated lipocalin (u-NGAL) levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the u-NGAL level is reduced by at least 5% compared to the u-NGAL level of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL level is reduced by at least 10% compared to the u-NGAL level of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL level is reduced by at least 20% compared to the u-NGAL level of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL level is reduced by at least 30%. In another embodiment, the u-NGAL level is reduced by at least 40% compared to the u-NGAL level of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced by at least 50% compared to the u-NGAL levels of patients transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced by at least 60% compared to the u-NGAL levels of patients transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced by at least 70% compared to the u-NGAL levels of patients transfused with conventionally stored blood. In yet another embodiment, the u-NGAL levels are reduced by at least 80% compared to the u-NGAL levels of patients transfused with conventionally stored blood. In a further embodiment, the u-NGAL levels are reduced by at least 90% compared to the u-NGAL levels of patients transfused with conventionally stored blood. In further aspects, u-NGAL levels are reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to u-NGAL levels in patients transfused with conventionally stored blood.

[0070] The disclosed method provides and includes reducing urinary neutrophil gelatinase-associated lipocalin (u-NGAL) levels in a patient in need of transfusion therapy comprising providing the patient 1.5-10 times oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the u-NGAL levels are reduced 2-3 times compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced 3-4 times compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced 4-10 times. In another embodiment, the u-NGAL levels are reduced 6-9 times compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In a further embodiment, the u-NGAL levels are reduced 2-5 times compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In another embodiment, u-NGAL levels are reduced 10-50 fold compared to u-NGAL levels in patients transfused with conventionally stored blood.

[0071] The disclosed method provides and includes reducing urinary neutrophil gelatinase-associated lipocalin (u-NGAL) levels in a patient in need of transfusion therapy, comprising providing the patient with at least 1.5 times oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the u-NGAL levels are reduced at least 2-fold compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced at least 3-fold compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced at least 4-fold compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In another embodiment, the u-NGAL levels are reduced at least 5-fold compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In a further embodiment, the u-NGAL levels are reduced at least 6-fold compared to the u-NGAL levels of a patient transfused with conventionally stored blood. In another embodiment, u-NGAL levels are reduced at least 7-fold compared to u-NGAL levels in patients transfused with conventionally stored blood. In another embodiment, u-NGAL levels are reduced at least 8-fold compared to u-NGAL levels in patients transfused with conventionally stored blood. In another embodiment, u-NGAL levels are reduced at least 9-fold. In another embodiment, u-NGAL levels are reduced at least 10-fold. In a further embodiment, u-NGAL levels are reduced at least 50-fold.

[0072] The disclosed method provides and includes reducing serum creatinine levels in patients who need transfusion therapy, comprising providing oxygen-reduced stored blood to the patient with oxygen saturation of 20% or less before and during storage. In one embodiment, serum creatinine levels are reduced by at least 5% compared to serum creatinine levels in patients transfused with conventionally stored blood. In another embodiment, serum creatinine levels are reduced by at least 10% compared to serum creatinine levels in patients transfused with conventionally stored blood. In another embodiment, serum creatinine levels are reduced by at least 20% compared to serum creatinine levels in patients transfused with conventionally stored blood. In another embodiment, serum creatinine levels are reduced by at least 30% compared to serum creatinine levels in patients transfused with conventionally stored blood. In another embodiment, serum creatinine levels are reduced by at least 40% compared to serum creatinine levels in patients transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 50% compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 60% compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 70% compared to the serum creatinine level of a patient transfused with conventionally stored blood. In yet another embodiment, the serum creatinine level is reduced by at least 80% compared to the serum creatinine level of a patient transfused with conventionally stored blood. In a further embodiment, the serum creatinine level is reduced by at least 90% compared to the serum creatinine level of a patient transfused with conventionally stored blood. In further aspects, serum creatinine levels are reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to serum creatinine levels in patients transfused with conventionally stored blood.

[0073] The disclosed method provides and includes reducing serum creatinine levels by 1.5-10 fold in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the serum creatinine level is reduced by 2-3 fold compared to serum creatinine levels in a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by 3-4 fold compared to serum creatinine levels in a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by 4-10 fold compared to serum creatinine levels in a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by 6-9 fold compared to serum creatinine levels in a patient transfused with conventionally stored blood. In a further embodiment, the serum creatinine level is reduced by 2-5 fold compared to serum creatinine levels in a patient transfused with conventionally stored blood. In another embodiment, serum creatinine levels are reduced 10-50 fold compared to serum creatinine levels in patients transfused with conventionally stored blood.

[0074] The disclosed method provides and includes reducing the serum creatinine level of a patient in need of transfusion therapy by at least 1.5-fold, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the serum creatinine level is reduced by at least 2-fold compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 3-fold compared to the AST level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 4-fold compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 5-fold. In a further embodiment, the serum creatinine level is reduced by at least 6-fold compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 7-fold compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 8-fold compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 9-fold compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced by at least 10-fold compared to the serum creatinine level of a patient transfused with conventionally stored blood.

[0075] The disclosed method provides and includes reducing serum creatinine levels to 0.5-1.5 mg / dL in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the serum creatinine level is reduced to 0.5-1 mg / dL relative to the serum creatinine level of a patient transfused with conventionally stored blood. In one embodiment, the serum creatinine level is reduced to 0.8-1 mg / dL relative to the serum creatinine level of a patient transfused with conventionally stored blood. In another embodiment, the serum creatinine level is reduced to 0.7-1.5 mg / dL relative to the serum creatinine level of a patient transfused with conventionally stored blood.

[0076] The disclosed method provides and includes reducing the serum creatinine level of a patient in need of transfusion therapy to less than 1.5 mg / dL, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the serum creatinine level is reduced to less than 1.4 mg / dL relative to the serum creatinine level of a patient transfused with conventionally stored blood. In one embodiment, the serum creatinine level is reduced to less than 1 mg / dL. In another embodiment, the serum creatinine level is reduced to less than 0.8 mg / dL relative to the serum creatinine level of a patient transfused with conventionally stored blood.

[0077] The disclosed method provides and includes reducing the patient's BUN level, comprising providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the BUN level is reduced by at least 5% compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 10% compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 20% compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 30% compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 40% compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 50% compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 60% compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 70% compared to the BUN level of a patient transfused with conventionally stored blood. In yet another embodiment, the BUN level is reduced by at least 80% compared to the BUN level of a patient transfused with conventionally stored blood. In a further embodiment, the BUN level is reduced by at least 90% compared to the BUN level of a patient transfused with conventionally stored blood. In a further embodiment, the BUN level is reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the BUN level of a patient transfused with conventionally stored blood.

[0078] The disclosed method provides and includes reducing the patient's BUN level by 1.5 to 10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the BUN level is reduced by 2 to 3 fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by 3 to 4 fold. In another embodiment, the BUN level is reduced by 4 to 10 fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by 6 to 9 fold compared to the BUN level of a patient transfused with conventionally stored blood. In a further embodiment, the BUN level is reduced by 2 to 5 fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by 10 to 100 fold compared to the BUN level of a patient transfused with conventionally stored blood.

[0079] The disclosed method provides and includes reducing the patient's BUN level by at least 1.5-fold, comprising providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the BUN level is reduced by at least 2-fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 3-fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 4-fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 5-fold compared to the BUN level of a patient transfused with conventionally stored blood. In a further embodiment, the BUN level is reduced by at least 6-fold. In another embodiment, the BUN level is reduced by at least 7-fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 8-fold compared to the BUN level of a patient transfused with conventionally stored blood. In another embodiment, the BUN level is reduced by at least 9-fold compared to the BUN level of a patient transfused with conventionally stored blood, hi another embodiment, the BUN level is reduced by at least 10-fold compared to the BUN level of a patient transfused with conventionally stored blood.

[0080] The disclosed method provides and includes reducing the percentage of CD45+ neutrophils in a patient, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the percentage of CD45+ neutrophils is reduced by at least 5% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 10% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 20% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 30% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 40% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 50% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 60% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 70% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by at least 80% compared to the CD45+ neutrophil level of a patient transfused with conventionally stored blood. In further embodiments, the percentage of CD45+ neutrophils is reduced by at least 90% compared to CD45+ neutrophil levels in patients transfused with conventionally stored blood. In further embodiments, the percentage of CD45+ neutrophils is reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to CD45+ neutrophil levels in patients transfused with conventionally stored blood.

[0081] The disclosed method provides and includes reducing the percentage of CD45+ neutrophils in a patient in need of transfusion therapy by 1.5-10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the percentage of CD45+ neutrophils is reduced by 2-3 fold compared to the CD45+ neutrophil levels of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by 3-4 fold compared to the CD45+ neutrophil levels of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by 4-10 fold compared to the CD45+ neutrophil levels of a patient transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by 6-9 fold compared to the CD45+ neutrophil levels of a patient transfused with conventionally stored blood. In a further embodiment, the percentage of CD45+ neutrophils is reduced 2-5 fold compared to CD45+ neutrophil levels in patients transfused with conventionally stored blood, hi another embodiment, the percentage of CD45+ neutrophils is reduced 10-50 fold compared to CD45+ neutrophil levels in patients transfused with conventionally stored blood.

[0082] The disclosed method provides and includes reducing the percentage of CD45+ neutrophils in trauma patients requiring 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 embodiment, the percentage of CD45+ neutrophils is reduced by at least 2-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 3-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 4-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 5-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In a further embodiment, the percentage of CD45+ neutrophils is reduced at least 6-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced at least 7-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced at least 8-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced at least 9-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced at least 10-fold compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood.

[0083] The disclosed method provides and includes reducing CXCL1 levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CXCL1 level is reduced by at least 5% compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 10% compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 20% compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 30% compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 40% compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 50% compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 60% compared to the CXCL1 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 70% compared to the CXCL1 level in a patient transfused with conventionally stored blood. In yet another embodiment, the CXCL1 level is reduced by at least 80% compared to the CXCL1 level in a patient transfused with conventionally stored blood. In a further embodiment, the CXCL1 level is reduced by at least 90% compared to the CXCL1 level in a patient transfused with conventionally stored blood. In a further embodiment, the CXCL1 level is reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the CXCL1 level in a patient transfused with conventionally stored blood.

[0084] The disclosed method provides and includes reducing CXCL1 levels in a patient by 1.5-10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CXCL1 levels are reduced by 2-3 fold compared to the CXCL1 levels in a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 levels are reduced by 3-4 fold compared to the CXCL1 levels in a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 levels are reduced by 4-10 fold compared to the CXCL1 levels in a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 levels are reduced by 6-9 fold compared to the CXCL1 levels in a patient transfused with conventionally stored blood. In a further embodiment, the CXCL1 levels are reduced by 2-5 fold compared to the CXCL1 levels in a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 levels are reduced by 10-100 fold compared to the CXCL1 levels in a patient transfused with conventionally stored blood.

[0085] The disclosed method provides and includes reducing the CXCL1 level of a patient by at least 1.5-fold, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CXCL1 level is reduced by at least 2-fold compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 3-fold compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 4-fold compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 5-fold compared to the CXCL1 level of a patient transfused with conventionally stored blood. In a further embodiment, the CXCL1 level is reduced by at least 6-fold compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 7-fold compared to the CXCL1 level of a patient transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 8-fold compared to the CXCL1 level in patients transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 9-fold compared to the CXCL1 level in patients transfused with conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least 10-fold compared to the CXCL1 level in patients transfused with conventionally stored blood.

[0086] Limited by the sensitivity of the test, healthy patients typically have a 5 × 10 -12The patient has an IL-6 level of less than 100 mg / L (pictograms / liter or pg / L). In the present disclosure, the patient in need has an IL-6 level of at least 5 pg / L. In another embodiment, the patient has an IL-6 level of 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 embodiment, the patient in need has an IL-6 level of 10-50 pg / L, 50-100 mg / L, 100-200 mg / L, or 10-200 mg / L. The method of the present disclosure provides and includes reducing IL-6 levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the IL-6 level is reduced by at least 5% compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 10% compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 20%. In another embodiment, the IL-6 level is reduced by at least 30% compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 40% compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 50% compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 60% compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 70% compared to the IL-6 level of a patient transfused with conventionally stored blood. In yet another embodiment, the IL-6 level is reduced by at least 80% compared to the IL-6 level of a patient transfused with conventionally stored blood. In a further embodiment, IL-6 levels are reduced by at least 90% as compared to IL-6 levels in patients transfused with conventionally stored blood.In further aspects, IL-6 levels are reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to IL-6 levels in patients transfused with conventionally stored blood.

[0087] The disclosed method provides and includes reducing IL-6 levels in a patient by 1.5 to 10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the IL-6 levels are reduced by 2 to 3 fold compared to the IL-6 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 levels are reduced by 3 to 4 fold compared to the IL-6 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 levels are reduced by 4 to 10 fold. In another embodiment, the IL-6 levels are reduced by 6 to 9 fold compared to the IL-6 levels of a patient transfused with conventionally stored blood. In a further embodiment, the IL-6 levels are reduced by 2 to 5 fold compared to the IL-6 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 levels are reduced by 10 to 100 fold compared to the IL-6 levels of a patient transfused with conventionally stored blood.

[0088] The disclosed method provides and includes reducing the IL-6 level of a patient by at least 1.5-fold, comprising providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the IL-6 level is reduced by at least 2-fold compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 3-fold compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 4-fold compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 5-fold. In a further embodiment, the IL-6 level is reduced by at least 6-fold compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 level is reduced by at least 7-fold compared to the IL-6 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-6 levels are reduced by at least 8-fold compared to the IL-6 levels of patients transfused with conventionally stored blood. In another embodiment, the IL-6 levels are reduced by at least 9-fold compared to the IL-6 levels of patients transfused with conventionally stored blood. In another embodiment, the IL-6 levels are reduced by at least 10-fold compared to the IL-6 levels of patients transfused with conventionally stored blood.

[0089] In one aspect of the disclosure, the patient in need is a patient with sepsis.

[0090] In one aspect of the disclosure, the patient in need thereof is a patient with 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.

[0091] In one embodiment of the present disclosure, the patient with inflammation is a patient with sickle cell disease. In a further embodiment, the patient with sickle cell disease has sickle cell anemia. In another embodiment, the patient with sickle cell disease has sickle cell crisis. In yet another embodiment, the patient has a type of sickle cell disease selected from the group consisting of hemoglobin SS, hemoglobin SC, hemoglobin SB+ beta thalassemia, hemoglobin SB (beta-0) thalassemia, hemoglobin SD, hemoglobin SE, and hemoglobin SO.

[0092] The present disclosure provides a method of preventing or reducing the number of vaso-occlusive episodes in sickle cell patients in need thereof, comprising providing pre-stored oxygen-reduced blood to a patient with sickle cell disease, the oxygen-reduced blood having an oxygen saturation of 20% or less during storage. In one embodiment, the vaso-occlusive episodes are reduced by at least 10% compared to sickle cell patients receiving conventionally stored blood. In another embodiment, 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.

[0093] The present disclosure provides for reducing red blood cell adhesion to endothelial cells expressing thrombospondin in a sickle cell patient in need thereof. Additionally, the present disclosure provides for reducing red blood cell adhesion to endothelial cells expressing vascular cell adhesion molecule-1 (VCAM-1) in a patient in need thereof. Further, the present disclosure provides for reducing red blood cell adhesion to endothelial cells expressing laminin in a patient in need thereof. In one embodiment, adhesion is reduced by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90% compared to adhesion of conventionally stored blood. In another embodiment, adhesion is reduced by 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90% compared to adhesion of conventionally stored blood.

[0094] The present disclosure provides a method of improving blood transfusion in a sickle cell patient in need, comprising providing a patient with sickle cell disease with pre-stored oxygen-reduced blood, the oxygen-reduced blood having an oxygen saturation of 20% or less during storage. In one embodiment, the patient has dactylitis (hand-foot syndrome). In another embodiment, the method of the present disclosure provides for reducing the occurrence of dactylitis. In another embodiment, the occurrence of dactylitis is reduced by at least 10, 20, 30, 40, 50, 60, 70, or 80% compared to patients receiving transfusions of conventionally stored RBCs. In another embodiment, the present disclosure provides for reducing the number of pain crises in a patient compared to patients treated with conventionally stored RBCs. In yet another embodiment, the present disclosure provides for reducing a complication of anemia selected from the group consisting of fatigue, irritability, dizziness, dyspnea, pale skin color, jaundice, growth retardation, and delayed puberty. In another embodiment, the present disclosure provides for reducing infections. In yet another embodiment, the present disclosure provides for preventing infections. In a further aspect, the present disclosure provides for reducing spleen damage.In one aspect, the spleen damage is spleen hemoglobin retention.In another aspect, the spleen damage is splenomegaly.In a further aspect, the present disclosure provides for reducing or preventing stroke.

[0095] In one embodiment of the disclosure, the patient in need is a patient with a fever. In another embodiment, the patient in need has a fever of at least 38°C. In another embodiment, the patient in need has a fever of about 39, 40, 41, or 42°C. In another embodiment, the patient has a fever of 38-42°C. In another embodiment, the patient in need has a fever and has increased c-reactive protein (CRP) levels compared to patients without a fever. In another embodiment, the patient with a fever has increased 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) compared to patients without a fever. In another embodiment, the patient with a fever has decreased levels of one or more inflammatory mediators selected from the group consisting of CCL-5 and CXCL-10 compared to patients without a fever.

[0096] In one embodiment of the disclosure, the patient in need is a patient at risk for fever. In another embodiment, the patient has a fever of 38-42°C. In another embodiment, the patient in need at risk for fever has an increased level of c-reactive protein (CRP) compared to a healthy individual. In another embodiment, the patient in need has an increased 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) compared to a healthy individual. In another embodiment, the patient in need has a decreased level of one or more inflammatory mediators selected from the group consisting of CCL-5 and CXCL-10 compared to a healthy patient.

[0097] Although limited by the sensitivity of the test, healthy patients typically have CRP levels below 3 mg / L. In one embodiment, the patient in need has a CRP level of at least 3 mg / L. In another embodiment, the patient has a CRP level of 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 embodiment, the patient in need has a CRP level of 3-5 mg / L, 3-10 mg / L, 5-10 mg / L, or 3-8 mg / L.

[0098] The disclosed method provides and includes reducing the CRP level of a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CRP level is restored to normal levels in healthy individuals. In one embodiment, the CRP level is reduced by at least 5% compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 10% compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 20%. In another embodiment, the CRP level is reduced by at least 30% compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 40% compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 50% compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 60% compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 70% compared to the CRP level of a patient transfused with conventionally stored blood. In yet another embodiment, the CRP level is reduced by at least 80% compared to the CRP level of a patient transfused with conventionally stored blood. In a further embodiment, the CRP level is reduced by at least 90% compared to the CRP level of a patient transfused with conventionally stored blood. In a further embodiment, the CRP level is reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the CRP level of a patient transfused with conventionally stored blood.

[0099] The disclosed method provides and includes reducing the CRP level of a patient in need by 1.5 to 10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CRP level is reduced by 2 to 3 fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by 3 to 4 fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by 4 to 10 fold. In another embodiment, the CRP level is reduced by 6 to 9 fold compared to the CRP level of a patient transfused with conventionally stored blood. In a further embodiment, the CRP level is reduced by 2 to 5 fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by 10 to 100 fold compared to the CRP level of a patient transfused with conventionally stored blood.

[0100] The disclosed method provides and includes reducing the CRP level of a patient in need of transfusion therapy by at least 1.5-fold, comprising providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CRP level is reduced by at least 2-fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 3-fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 4-fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 5-fold. In a further embodiment, the CRP level is reduced by at least 6-fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 7-fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 8-fold compared to the CRP level of a patient transfused with conventionally stored blood. In another embodiment, the CRP level is reduced by at least 9-fold compared to the CRP level in a patient transfused with conventionally stored blood, hi another embodiment, the CRP level is reduced by at least 10-fold compared to the CRP level in a patient transfused with conventionally stored blood.

[0101] Although limited by the sensitivity of the test, healthy patients typically have IL-8 levels below 40 picograms per liter (pg / L). In some embodiments, the patient in need has an IL-8 level of at least 40 pg / L. In other embodiments, the patient has an IL-8 level of 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 other embodiments, the patient in need has a CRP level of 30-50 pg / L, 40-100 pg / L, 50-100 pg / L, or 60-200 pg / L.

[0102] The disclosed method provides and includes reducing IL-8 levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, IL-8 levels are restored to normal levels in a healthy individual. In one embodiment, IL-8 levels are reduced by at least 5% compared to IL-8 levels in a patient transfused with conventionally stored blood. In another embodiment, IL-8 levels are reduced by at least 10% compared to IL-8 levels in a patient transfused with conventionally stored blood. In another embodiment, IL-8 levels are reduced by at least 20%. In another embodiment, IL-8 levels are reduced by at least 30% compared to IL-8 levels in a patient transfused with conventionally stored blood. In another embodiment, IL-8 levels are reduced by at least 40% compared to IL-8 levels in a patient transfused with conventionally stored blood. In another embodiment, IL-8 levels are reduced by at least 50% compared to IL-8 levels in a patient transfused with conventionally stored blood. In another embodiment, the IL-8 level is reduced by at least 60% compared to the IL-8 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 level is reduced by at least 70% compared to the IL-8 level of a patient transfused with conventionally stored blood. In yet another embodiment, the IL-8 level is reduced by at least 80% compared to the IL-8 level of a patient transfused with conventionally stored blood. In a further embodiment, the IL-8 level is reduced by at least 90% compared to the IL-8 level of a patient transfused with conventionally stored blood. In a further embodiment, the IL-8 level is reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the IL-8 level of a patient transfused with conventionally stored blood.

[0103] The disclosed method provides and includes reducing IL-8 levels in a patient in need thereof by 1.5 to 10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the IL-8 levels are reduced by 2 to 3 fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced by 3 to 4 fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced by 4 to 10 fold. In another embodiment, the IL-8 levels are reduced by 6 to 9 fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In a further embodiment, the IL-8 levels are reduced by 2 to 5 fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced by 10 to 100 fold compared to the IL-8 levels of a patient transfused with conventionally stored blood.

[0104] The disclosed method provides and includes reducing IL-8 levels in a patient in need thereof by at least 1.5-fold, comprising providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the IL-8 levels are reduced by at least 2-fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced by at least 3-fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced by at least 4-fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced by at least 5-fold. In a further embodiment, the IL-8 levels are reduced by at least 6-fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced by at least 7-fold compared to the IL-8 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced at least 8-fold compared to the IL-8 levels of patients transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced at least 9-fold compared to the IL-8 levels of patients transfused with conventionally stored blood. In another embodiment, the IL-8 levels are reduced at least 10-fold compared to the IL-8 levels of patients transfused with conventionally stored blood.

[0105] Healthy individuals have IL-10 levels of 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, a patient in need thereof has an IL-10 level of at least 15 pg / L. In another embodiment, a patient has an IL-10 level of 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 embodiment, a patient in need thereof has an IL-10 level of 20-50 pg / L, 50-100 mg / L, 100-200 mg / L, or 15-200 mg / L.

[0106] The disclosed method provides and includes reducing IL-10 levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, IL-10 levels are restored to normal levels in a healthy individual. In one embodiment, IL-10 levels are reduced by at least 5% compared to IL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, IL-10 levels are reduced by at least 10% compared to IL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, IL-10 levels are reduced by at least 20%. In another embodiment, IL-10 levels are reduced by at least 30% compared to IL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, IL-10 levels are reduced by at least 40% compared to IL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, the IL-10 level is reduced by at least 50% compared to the IL-10 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 level is reduced by at least 60% compared to the IL-10 level of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 level is reduced by at least 70% compared to the IL-10 level of a patient transfused with conventionally stored blood. In yet another embodiment, the IL-10 level is reduced by at least 80% compared to the IL-10 level of a patient transfused with conventionally stored blood. In a further embodiment, the IL-10 level is reduced by at least 90% compared to the IL-10 level of a patient transfused with conventionally stored blood. In further aspects, IL-10 levels are reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to IL-10 levels in patients transfused with conventionally stored blood.

[0107] The disclosed method provides and includes reducing IL-10 levels in a patient in need thereof by 1.5-10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the IL-10 levels are reduced by 2-3 fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced by 3-4 fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced by 4-10 fold. In another embodiment, the IL-10 levels are reduced by 6-9 fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In a further embodiment, the IL-10 levels are reduced by 2-5 fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced by 10-100 fold compared to the IL-10 levels of a patient transfused with conventionally stored blood.

[0108] The disclosed method provides and includes reducing IL-10 levels in a patient in need thereof by at least 1.5-fold, comprising providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the IL-10 levels are reduced by at least 2-fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced by at least 3-fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced by at least 4-fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced by at least 5-fold. In a further embodiment, the IL-10 levels are reduced by at least 6-fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced by at least 7-fold compared to the IL-10 levels of a patient transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced at least 8-fold compared to the IL-10 levels of patients transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced at least 9-fold compared to the IL-10 levels of patients transfused with conventionally stored blood. In another embodiment, the IL-10 levels are reduced at least 10-fold compared to the IL-10 levels of patients transfused with conventionally stored blood.

[0109] The disclosed method provides and includes reducing G-CSF levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the G-CSF level is restored to normal levels in a healthy person. In one embodiment, the G-CSF level is reduced by at least 5% compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 10% compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 20%. In another embodiment, the G-CSF level is reduced by at least 30% compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 40% compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 50% compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 60% compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 70% compared to the G-CSF level of a patient transfused with conventionally stored blood. In yet another embodiment, the G-CSF level is reduced by at least 80% compared to the G-CSF level of a patient transfused with conventionally stored blood. In a further embodiment, the G-CSF level is reduced by at least 90% compared to the G-CSF level of a patient transfused with conventionally stored blood. In further embodiments, G-CSF levels are reduced by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to G-CSF levels in patients transfused with conventionally stored blood.

[0110] The disclosed method provides and includes reducing the G-CSF level of a patient by 1.5 to 10 fold, including providing the patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the G-CSF level is reduced by 2 to 3 fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by 3 to 4 fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by 4 to 10 fold. In another embodiment, the G-CSF level is reduced by 6 to 9 fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In a further embodiment, the G-CSF level is reduced by 2 to 5 fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by 10 to 100 fold compared to the G-CSF level of a patient transfused with conventionally stored blood.

[0111] The disclosed method provides and includes reducing the G-CSF level of a patient in need of transfusion therapy by at least 1.5-fold, comprising providing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the G-CSF level is reduced by at least 2-fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 3-fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 4-fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 5-fold. In a further embodiment, the G-CSF level is reduced by at least 6-fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 7-fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 8-fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 9-fold compared to the G-CSF level of a patient transfused with conventionally stored blood. In another embodiment, the G-CSF level is reduced by at least 10-fold compared to the G-CSF level of a patient transfused with conventionally stored blood.

[0112] The disclosed method provides and includes increasing CCL-5 levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-enhanced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, CCL-5 levels are restored to normal levels in a healthy individual. In one embodiment, CCL-5 levels are increased by at least 5% compared to CCL-5 levels in a patient transfused with conventionally stored blood. In another embodiment, CCL-5 levels are increased by at least 10% compared to CCL-5 levels in a patient transfused with conventionally stored blood. In another embodiment, CCL-5 levels are increased by at least 20%. In another embodiment, CCL-5 levels are increased by at least 30% compared to CCL-5 levels in a patient transfused with conventionally stored blood. In another embodiment, CCL-5 levels are increased by at least 40% compared to CCL-5 levels in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased by at least 50% compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased by at least 60% compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased by at least 70% compared to the CCL-5 level in a patient transfused with conventionally stored blood. In yet another embodiment, the CCL-5 level is increased by at least 80% compared to the CCL-5 level in a patient transfused with conventionally stored blood. In a further embodiment, the CCL-5 level is increased by at least 90% compared to the CCL-5 level in a patient transfused with conventionally stored blood. In further aspects, CCL-5 levels are increased by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to CCL-5 levels in patients transfused with conventionally stored blood.

[0113] The disclosed method provides and includes increasing CCL-5 levels in a patient in need thereof by 1.5 to 10 fold, including providing the patient with oxygen-enhanced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CCL-5 levels are reduced by 2 to 3 fold compared to the CCL-5 levels of a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 levels are increased by 3 to 4 fold compared to the CCL-5 levels of a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 levels are increased by 4 to 10 fold. In another embodiment, the CCL-5 levels are increased by 6 to 9 fold compared to the CCL-5 levels of a patient transfused with conventionally stored blood. In a further embodiment, the CCL-5 levels are increased by 2 to 5 fold compared to the CCL-5 levels of a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 levels are increased by 10 to 100 fold compared to the CCL-5 levels of a patient transfused with conventionally stored blood.

[0114] The disclosed method provides and includes increasing CCL-5 levels in a patient in need of transfusion therapy by at least 1.5-fold, comprising providing oxygen-enhanced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the CCL-5 level is increased by at least 2-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased by at least 3-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased by at least 4-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased by at least 5-fold. In a further embodiment, the CCL-5 level is increased by at least 6-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased by at least 7-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased at least 8-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased at least 9-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood. In another embodiment, the CCL-5 level is increased at least 10-fold compared to the CCL-5 level in a patient transfused with conventionally stored blood.

[0115] The disclosed method provides and includes increasing CXCL-10 levels in a patient in need of transfusion therapy, comprising providing the patient with oxygen-enhanced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, CXCL-10 levels are restored to normal levels in a healthy individual. In one embodiment, CXCL-10 levels are increased by at least 5% compared to CXCL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, CXCL-10 levels are increased by at least 10% compared to CXCL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, CXCL-10 levels are increased by at least 20%. In another embodiment, CXCL-10 levels are increased by at least 30% compared to CXCL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, CXCL-10 levels are increased by at least 40% compared to CXCL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased by at least 50% compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased by at least 60% compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased by at least 70% compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In yet another embodiment, the CXCL-10 level is increased by at least 80% compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In a further embodiment, the CXCL-10 level is increased by at least 90% compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In further aspects, CXCL-10 levels are increased by 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to CXCL-10 levels in patients transfused with conventionally stored blood.

[0116] The disclosed method provides and includes increasing CXCL-10 levels in a patient in need thereof by 1.5-10 fold, comprising providing the patient with oxygen-enhanced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CXCL-10 levels are reduced by 2-3 fold compared to the CXCL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 levels are increased by 3-4 fold compared to the CXCL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 levels are increased by 4-10 fold. In another embodiment, the CXCL-10 levels are increased by 6-9 fold compared to the CXCL-10 levels in a patient transfused with conventionally stored blood. In a further embodiment, the CXCL-10 levels are increased by 2-5 fold compared to the CXCL-10 levels in a patient transfused with conventionally stored blood. In another embodiment, CXCL-10 levels are increased 10-100 fold compared to CXCL-10 levels in patients transfused with conventionally stored blood.

[0117] The disclosed method provides and includes increasing CXCL-10 levels in a patient in need of transfusion therapy by at least 1.5-fold, comprising providing oxygen-enhanced stored blood having an oxygen saturation of 20% or less before and during storage to a patient in need of transfusion therapy. In one embodiment, the CXCL-10 level is increased by at least 2-fold compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased by at least 3-fold compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased by at least 4-fold compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased by at least 5-fold. In a further embodiment, the CXCL-10 level is increased by at least 6-fold compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased by at least 7-fold compared to the CXCL-10 level in a patient transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased at least 8-fold compared to the CXCL-10 level in patients transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased at least 9-fold compared to the CXCL-10 level in patients transfused with conventionally stored blood. In another embodiment, the CXCL-10 level is increased at least 10-fold compared to the CXCL-10 level in patients transfused with conventionally stored blood.

[0118] The terms "higher," "greater," or "increased" as used herein mean that measurements of anaerobically stored blood with reduced oxygen are at least one standard deviation greater when compared to measurements of otherwise identically treated conventionally stored blood, with a sample size of at least two for each comparative measurement condition.

[0119] The terms "reduce," "reduced," "lower," "reduced," or "less" as used herein mean that measurements of oxygen-reduced anaerobically stored blood are at least one standard deviation lower with a sample size of at least 5 for each comparative measurement condition when compared to measurements of otherwise identically treated normoxic or hyperoxic conventionally stored blood RBCs.

[0120] The term "about" as used herein refers to ±10%.

[0121] The term "less than" as used herein refers to a lesser amount as well as an amount greater than zero.

[0122] The terms "comprises," "comprising," "includes," "including," "having," and their conjugations mean "including but not limited to."

[0123] The term "consisting of" means "including but not limited to."

[0124] The term "consisting essentially of" means that a composition, method, or structure may include additional components, steps, and / or moieties, but only if the additional components, steps, and / or moieties do not materially alter the basic and novel characteristics of the claimed composition, method, or structure.

[0125] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0126] The term "blood" as used herein refers to whole blood, leukocyte-depleted RBCs, platelet-depleted RBCs, and leukocyte- and platelet-depleted RBCs. The term blood further includes packed red blood cells, platelet-depleted packed red blood cells, leukocyte-depleted packed red blood cells, and leukocyte- and platelet-depleted packed red blood cells. The temperature of blood may vary depending on the stage of the collection process, initially at the time and point of collection, at normal body temperature of 37°C, but rapidly drops to about 30°C as soon as the blood leaves the patient's body, and then further drops to room temperature in about 6 hours when unprocessed, and finally refrigerated at about 4°C to 6°C. In vivo human red blood cells are in a dynamic state. Red blood cells contain hemoglobin, an iron-containing protein that carries oxygen throughout the body and gives blood its red color. The percentage of blood volume that is composed of red blood cells is called the hematocrit. As used herein, unless otherwise limited, RBCs also include packed red blood cells (pRBCs). Packed red blood cells are prepared from whole blood using centrifugation techniques commonly known in the art. As used herein, unless otherwise indicated, the hematocrit of pRBC is about 70%. As used herein, oxygen-reduced stored RBCs can include oxygen and carbon dioxide reduced stored RBCs. As used herein, oxygen-reduced (OR) blood can include oxygen and carbon dioxide (OCR) reduced blood.

[0127] As used herein, the terms "patient" and "subject" are used interchangeably to mean a person or animal in need of a blood transfusion.

[0128] The term "trauma" as used herein includes blood loss, hemorrhagic trauma.

[0129] The term "hemorrhagic shock" as used herein is shock resulting from loss of circulatory blood volume and / or oxygen-carrying capacity. Hemorrhagic shock results from any condition associated with blood loss, internal (e.g., gastrointestinal) or external bleeding, and trauma (e.g., penetrating or blunt trauma), among others.

[0130] The term "adverse event" as used herein includes events resulting from hemorrhagic shock in hemorrhagic trauma patients.

[0131] The terms "injury," "damage," and "failure" as used herein refer to an organ that does not function properly or does not function as expected in a person or animal that does not have the disease or injury.

[0132] As used herein, a "unit" of blood is approximately 450-500 ml containing an anticoagulant. Suitable anticoagulants include Citrate Phosphate Dextrose Solution (CPD), Citrate Phosphate Dextrose Adenine (CPDA1), Acid Citrate Dextrose (ACD), and Anticoagulant Citrate Dextrose Solution Solution A (ACD-A).

[0133] Throughout this application, various aspects of the disclosure may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Thus, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values ​​within that range. For example, the description of a range such as "1-6" should be considered to have specifically disclosed subranges within that range such as "1-3", "1-4", "1-5", "2-4", "2-6", "3-6", and individual numbers such as 1, 2, 3, 4, 5, and 6. This is true regardless of the breadth of the range.

[0134] Whenever a numerical range is given herein, it is meant to include any recited number (fractional or integer) within the given range. The phrases "range / range between" a first indicated number and a second indicated number, and "range / range from" a first indicated number to a second indicated number, are used interchangeably herein and are meant to include the first and second indicated numbers and all fractional and integer numbers therebetween.

[0135] The term "method" as used herein refers to ways, means, techniques, and procedures for accomplishing a given task, including, but not limited to, providing oxygen-reduced stored blood to a human patient in need of a transfusion, the blood having an initial oxygen saturation of 20% or less and that has been stored for at least two days.

[0136] The present disclosure provides the following embodiments. Embodiment 1. A method for treating low mean arterial pressure in a subject in need thereof, comprising providing stored oxygen-reduced blood to a subject having low mean arterial pressure resulting from hemorrhagic trauma, said oxygen-reduced blood having an initial oxygen saturation of 20% or less and maintained at an oxygen saturation of 20% or less during a storage period. Embodiment 2. The method of embodiment 1, wherein the mean arterial pressure in the subject in need is increased compared to a patient receiving conventionally stored blood following the donation. Embodiment 3. The method of embodiment 1 or 2, wherein the bleeding trauma is selected from the group consisting of surgery, penetrating wounds, blunt force trauma, fall injuries, and motor vehicle accident injuries. Embodiment 4. The method of any one of embodiments 1-3, wherein the increase in mean arterial pressure in the subject in need is at a rate that is greater than the rate of increase in mean arterial pressure in a subject not receiving the stored oxygen-reduced blood. Embodiment 5. The method of any one of embodiments 1-4, wherein the mean arterial pressure in the subject in need is increased by at least 20% after the donation compared to the mean arterial pressure of a patient transfused with conventionally stored blood. Embodiment 6. The method of any one of embodiments 1-5, wherein the mean arterial pressure in the subject remains increased by at least 20% for at least 1 hour after the providing. Embodiment 7. The method of any one of embodiments 1 to 5, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 8. A method for reducing the amount of blood required for transfusion in a trauma patient in need thereof, comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 9. The method of embodiment 8, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 10. A method for reducing hemorrhagic shock in a trauma patient in need thereof, comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage, said trauma patient comprising a lactate level of 0.5-2.5 mmol / L before said providing, and said hemorrhagic shock is reversed. Embodiment 11. The method of embodiment 10, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 12. A method of reducing liver injury in a trauma patient requiring transfusion therapy comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 13. The method of embodiment 9, wherein the trauma patient has improved aspartate aminotransferase (AST) levels, alanine aminotransferase (ALT) levels, or a combination thereof after said providing. Embodiment 14. The method of embodiment 13, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 15. A method of reducing renal failure in a trauma patient requiring transfusion therapy comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 16. The method of embodiment 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. Embodiment 17. The method of embodiment 15 or 16, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 18. A method of reducing lung injury in a trauma patient requiring transfusion therapy comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 19. The method of embodiment 18, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 20. A method of reducing lactate in a trauma patient in need thereof comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 21. The method of embodiment 20, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 22. A method of reducing aspartate aminotransferase (AST) in a trauma patient in need thereof comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 23. A method of reducing alanine aminotransferase (ALT) in a trauma patient in need thereof comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 24. A method of reducing blood urea nitrogen (BUN) levels in a trauma patient in need thereof comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 25. A method of reducing neutrophil gelatinase-associated lipocalin (NGAL) in a trauma patient in need thereof comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 26. A method of reducing serum creatinine in a trauma patient in need thereof comprising providing reduced-oxygen blood having an oxygen saturation of 20% or less before and during storage. Embodiment 27. The method of any one of embodiments 22-26, wherein the blood is oxygen-reduced and carbon dioxide-reduced stored blood. Embodiment 28. Use of donor blood to produce reduced oxygen blood having an oxygen saturation of 20% or less before and during storage for therapeutic applications of low mean arterial pressure. Embodiment 29. Use of donor blood to produce oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage for therapeutic applications in reducing adverse events in hemorrhagic trauma patients, said adverse events being selected from the group including liver injury, pulmonary failure, renal failure, and cardiac failure. Embodiment 30. The use of embodiment 28 or 29, wherein the preparation comprises oxygen and carbon dioxide reduced blood. Embodiment 31. A method of treating a patient in need of treatment with stored oxygen-depleted blood having an oxygen saturation of 20% or less during storage, wherein the patient in need has inflammation. Embodiment 32. The method of embodiment 31, wherein the inflammation is chronic or acute. Embodiment 33. The method of embodiment 31, wherein the inflammation is systemic. Embodiment 34. The method of any one of embodiments 31-33, wherein the inflammation is due to trauma, infection, cancer, coagulation disorder, or autoimmunity. Embodiment 35. The method of any one of embodiments 31-34, wherein the patient further has or is at risk of developing a fever. Embodiment 36. The method of embodiment 35, wherein the fever is at least 38° C. Embodiment 37. The method of any one of embodiments 31-36, wherein the patient has elevated C-reactive protein (CRP) levels. Embodiment 38. The method of embodiment 37, wherein the patient has a CRP level of at least 2.0 mg / L. Embodiment 39. The method of embodiment 38, wherein the patient has a CRP level of at least 3.0 mg / L. Embodiment 40. The method of any one of embodiments 37-39, wherein the patient has a CRP level of 3.0 to 10 mg / L. Embodiment 41. The method of any one of embodiments 31-40, wherein the patient has increased 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) compared to patients without fever. Embodiment 42. The method of embodiment 41, wherein the patient has an IL-6 level of at least 5 pg / mL. Embodiment 43. The method of embodiment 41, wherein the patient has an IL-8 level of at least 29 pg / mL. Embodiment 44. The method of embodiment 41, wherein the patient has an IL-10 level of at least 15 pg / mL. Embodiment 45. The method of embodiment 41, wherein the patient has a G-CSF level of at least 35 pg / mL. Embodiment 46. The method of any one of embodiments 31-45, wherein the patient has reduced levels of one or more inflammatory mediators selected from the group consisting of CCL5 and CXCL10 compared to a patient without fever. Embodiment 47. The method of embodiment 46, wherein the patient has a CCL5 level of at least 380 pg / mL. Embodiment 48. The method of embodiment 46, wherein the patient has a CXCL10 level of at least 270 pg / mL. Embodiment 49. The method of any one of embodiments 31-48, wherein the patient has liver damage. Embodiment 50. The method of any one of embodiments 31-49, wherein the patient has a lung injury. Embodiment 51. The method of any one of embodiments 31-50, wherein the patient has a splenic injury. Embodiment 52. The method of any one of embodiments 31-51, wherein the patient has kidney damage. Embodiment 53. The method of any one of embodiments 31-52, wherein the patient has a bone injury. Embodiment 54. The method of any one of embodiments 31-53, wherein the treatment reduces adhesion of red blood cells to thrombospondin in the patient compared to patients treated with conventionally stored blood. Embodiment 55. The method of embodiment 54, wherein the thrombospondin is reduced by at least 10%. Embodiment 56. The method of any one of embodiments 31-55, wherein the stored oxygen-reduced blood is oxygen and carbon dioxide-reduced blood. Embodiment 57. A method of improving blood transfusions in a sickle cell patient in need thereof, comprising providing stored oxygen-reduced blood to a patient with sickle cell disease, said oxygen-reduced blood having an oxygen saturation of 20% or less during storage. Embodiment 58. The method of embodiment 57, wherein the sickle cell disease is sickle cell anemia. Embodiment 59. The method of embodiment 57, wherein the sickle cell disease is sickle cell crisis. Embodiment 60. The method of embodiment 57, wherein said sickle cell disease is selected from the group consisting of hemoglobin SS (HbSS), hemoglobin SC (HbSC), hemoglobin S beta thalassemia+ (HbSB+), hemoglobin S (beta-0) thalassemia (HbSB), hemoglobin SD (HbSD), hemoglobin SE (HbSE), and hemoglobin SO (HbSO). Embodiment 61. The method of any one of embodiments 57-60, wherein the improvement comprises reducing the incidence of dactylitis (hand-foot syndrome) by at least 10% compared to patients receiving conventionally stored blood. Embodiment 62. The method of any one of embodiments 57-61, wherein the improvement comprises reducing the number of pain crises by at least 10% compared to being treated with conventionally stored blood. Embodiment 63. The method of any one of embodiments 57-62, wherein the amelioration comprises reducing a complication of anemia selected from the group consisting of fatigue, irritability, dizziness, dyspnea, pale skin, jaundice, growth retardation, and delayed puberty. Embodiment 64. The method of any one of embodiments 57-63, wherein the amelioration comprises reducing the incidence of infection. Embodiment 65. The method of any one of embodiments 57-64, wherein the improvement comprises reducing splenic damage. Embodiment 66. The method of embodiment 65, wherein the splenic injury is splenic erythrocyte retention. Embodiment 67. The method of embodiment 65, wherein the splenic injury is splenic enlargement. Embodiment 68. The method of any one of embodiments 57-67, wherein the improving comprises reducing or preventing stroke. Embodiment 69. The method of any one of embodiments 57-68, wherein the pre-stored oxygen-reduced blood has a higher 24-hour recovery rate after the transfusion compared to conventionally stored blood. Embodiment 70. The method of any one of embodiments 57-69, wherein the pre-stored oxygen-reduced blood has increased deformability compared to conventionally stored blood. Embodiment 71. The method of any one of embodiments 57-70, wherein the oxygen-reduced blood has increased deformability in the presence of sickle cell plasma compared to conventionally stored blood. Embodiment 72. The method of any one of embodiments 57-71, further comprising reducing carbon dioxide in the oxygen-reduced blood. Embodiment 73. A method of reducing a number of vaso-occlusive episodes in a patient in need thereof, comprising providing stored oxygen-reduced blood to a patient in need thereof, said oxygen-reduced blood having an oxygen saturation of 20% or less during storage, and wherein reducing said vaso-occlusive episodes comprises reducing adhesion of red blood cells to endothelial cells. Embodiment 74. The method of embodiment 73, wherein the patient in need thereof has sickle cell disease. Embodiment 75. The method of embodiment 73 or 74, wherein the patient in need thereof has inflammation. Embodiment 76. The method of any one of embodiments 73-75, wherein the reducing adhesion of red blood cells is to endothelial cells that express thrombospondin. Embodiment 77. The method of any one of embodiments 73-76, wherein the reducing adhesion of red blood cells is to endothelial cells expressing vascular cell adhesion molecule-1 (VCAM-1). Embodiment 78. The method of any one of embodiments 73-77, wherein the reducing adhesion of red blood cells is to endothelial cells expressing laminin. Embodiment 79. The method of any one of embodiments 76-78, wherein the adhesion is reduced by at least 10% compared to conventionally stored blood. Embodiment 80. The method of any one of embodiments 76-79, wherein said adhesion is reduced by at least 20% compared to conventionally stored blood. Embodiment 81. The method of any one of embodiments 76-80, wherein the adhesion is reduced by 10-50%. Embodiment 82. The method of any one of embodiments 73-81, wherein the sickle cell disease is sickle cell anemia. Embodiment 83. The method of any one of embodiments 73-81, wherein the sickle cell disease is sickle cell crisis. Embodiment 84. The method of any one of embodiments 73-83, wherein the sickle cell disease is selected from the group consisting of hemoglobin SS (HbSS), hemoglobin SC (HbSC), hemoglobin S beta thalassemia+ (HbSB+), hemoglobin S (beta-0) thalassemia (HbSB), hemoglobin SD (HbSD), hemoglobin SE (HbSE), and hemoglobin SO (HbSO). Embodiment 85. The method of any one of embodiments 73-83, further comprising reducing carbon dioxide in the oxygen-reduced blood. Embodiment 86. A method of reducing adhesion of red blood cells to endothelial cells in a patient in need thereof, comprising providing stored oxygen-reduced blood to a patient having sickle cell disease, said oxygen-reduced blood having an oxygen saturation of 20% or less during storage. Embodiment 87. Oxygen-reduced blood having an oxygen saturation of 20% or less during storage for use in the treatment of sickle cell disease. Embodiment 88. The oxygen-reduced blood of embodiment 87, wherein the blood is oxygen and carbon dioxide depleted blood. Embodiment 89. Deoxygenated blood having an oxygen saturation of 20% or less during storage for use in treating inflammation. Embodiment 90. The oxygen-reduced blood of embodiment 89, wherein the blood is oxygen and carbon dioxide-reduced blood. Embodiment 91. Stored deoxygenated blood comprising an oxygen saturation of 20% or less for use in treating vaso-occlusive episodes, said treatment reducing the number of vaso-occlusive episodes in a patient in need thereof, said reducing vaso-occlusive episodes comprising reducing adhesion of red blood cells to endothelial cells. Embodiment 92. The stored oxygen-reduced blood of embodiment 91, wherein the blood is oxygen and carbon dioxide reduced stored blood. Embodiment 93. Use of deoxygenated blood having an oxygen saturation of 20% or less during storage for the treatment of inflammation in a patient in need thereof. Embodiment 94. Use of oxygen-reduced blood having an oxygen saturation of 20% or less during storage for the treatment of sickle cell in a patient in need thereof. Embodiment 95. The use of oxygen-reduced blood according to embodiment 94, wherein the blood is oxygen and carbon dioxide-reduced blood. Embodiment 96. Use of deoxygenated blood substantially as hereinbefore shown and described. Embodiment 97. Use of oxygen and carbon dioxide reduced blood substantially as hereinbefore shown and described. Embodiment 98. A kit comprising an oxygen-impermeable blood storage bag containing stored oxygen-reduced blood having an oxygen saturation of 20% or less during storage, a filter, and a drip chamber. Embodiment 99. The kit of embodiment 98, further comprising a device for adding gas to stored oxygen-reduced blood prior to transfusion. Embodiment 100. The kit of embodiment 98 or 99, further comprising tubing for transfusing the stored oxygen-reduced blood to a patient in need thereof. Embodiment 101. Use of the kit according to any one of embodiments 98 to 100 for the treatment of sickle cell disease or inflammation in a patient in need thereof.

[0137] Although the disclosure has been described with reference to specific embodiments, those skilled in the art will recognize that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the disclosure. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the disclosure without departing from its scope.

[0138] Therefore, it is intended that this disclosure not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this disclosure, but that the disclosure will include all embodiments falling within the scope and spirit of the appended claims. [Example] EXAMPLES

[0139] Blood collection and sample preparation Collect each pool of red blood cells from a total of 12-14 rats in CP2D anticoagulant. The pooled blood is leukoreduced using a neonatal leukoreduction filter, the components are separated, and the RBCs are stored in AS-3 additive solution. Collect a total of two RBC pools. Split each pool into four: untreated control (C), sham control (SC), oxygen reduced (OR), and oxygen and carbon dioxide reduced (OCR). For the C, SC, OR, and OCR units, process the RBC subunits by transferring them into 80 mL PVC blood transport bags, and the final RBC product is made by a gas exchange process. Fill the RBC bags, except for C, with 100% N2 (for OR) or 95% N2 / 5% CO2 (for OCR) or air (SC) through a sterile filter and gently rotate the long side at 2-3 RPM (except C). For the OR and OCR units, remove the gas through the filter after 10 min and introduce fresh gas for the subsequent gas exchange process. This process is repeated 5-8 times until a target %SO2 of 5-10% is achieved as measured by an ABL-90 co-oximeter (Radiometer Copenhagen). The SC units are rotated for 60 min without gas exchange. OR and OCR units are stored anaerobically in N2-filled canisters, while C and SC units are stored in ambient air. All units are stored at 4 °C for 3 weeks and sampled on days 0 or 1, 7, 14, 21, and 28. Two pools were prepared and stored.

[0140] ATP, 2,3-DPG, and hemolysis analyses are performed on days 0, 1, 7, 14, 21, and 28. As shown in FIG. 1, ATP levels are higher in OR blood on day 21 and OCR blood on days 7, 14, 21, and 28 compared to conventionally stored blood (control). In addition, OR blood has higher levels of 2.3-DPG on days 2, 7, and 14 compared to the control. In addition, OCR blood shows higher levels of 2,3-DPG on days 2, 7, and 14 compared to the control. See FIG. 2. EXAMPLES

[0141] Recovery rate of oxygen-reduced blood Small volumes (<200 μL) of control, OR, and OCR blood stored for 3 weeks are labeled with technetium-99m. Animals are transfused with labeled RBCs (<200 uL) and circulating radioactivity is measured periodically up to 24 h to estimate the percentage of transfused RBCs that survive 24 h post-transfusion. As shown in Figure 3, when RBCs are stored for 3 weeks, significantly more OR-RBCs and OCR-RBCs are recovered compared to control RBCs. EXAMPLES

[0142] A rat model of hemorrhagic shock resuscitation Blood collection and sample preparation. Collect each pool of red blood cells from a total of 12-14 rats into CP2D anticoagulant. The pooled blood is leukoreduced using a neonatal leukoreduction filter, components are separated, and the RBCs are stored in AS-3 additive solution. Collect a total of six RBC pools. Prepare two pools for conventional storage (control). Deplete oxygen from two pools (oxygen reduced; OR) and deplete oxygen and carbon dioxide from the remaining two blood pools (oxygen and carbon dioxide reduced; OCR). Each of the four pools to be reduced is processed by transferring the RBCs into a 600 mL PVC blood transport bag, and the final RBC product is created by a gas exchange process. Fill the RBC bag with 100% N2 (for OR) or 95% N2 / 5% CO2 (for OCR) through a sterile filter and gently rotate the long side at 60-90 RPM. Remove the gas through the filter after 10 min and introduce fresh gas for the subsequent gas exchange process. Repeat this process 5-8 times until a target %SO2 of 5-10% is achieved as measured by an ABL-90 co-oximeter (Radiometer Copenhagen). Store OR and OCR blood anaerobically in N2-filled canisters.

[0143] Studies are performed in Sprague-Dawley rats (Charles River Laboratories, Boston, MA) weighing 150-200 grams (g). Briefly, animals are anesthetized with 40 mg / kg sodium pentobarbital administered intraperitoneally. Core body temperature is maintained at 37°C by placing the animal in a supine position on a heating pad. Animals are prepared as follows: (i) left jugular vein and left femoral artery catheterization, (ii) tracheotomy (polyethylene-90 tubing), and (iii) left ventricular (LV) conductance catheterization through the right carotid artery. Animals are mechanically ventilated (TOPO ventilator, Kent Scientific, Torrington, CT) using room air at a respiratory rate of 50-70 breaths per minute and a peak inspiratory pressure of 10-15 cmH2O. After instrumentation, administer volatile anesthesia (1.5% / vol isoflurane, Dragerwerk AG, Laubeck, Germany) using a vaporizer connected to the ventilator. Continuously check the depth of anesthesia by toe pinch and increase the isoflurane by 0.1% / vol if necessary to prevent animal discomfort.

[0144] Anesthetized animals are bled by withdrawing 50% of the animal's blood volume (BV; estimated 7% of body weight) via the femoral artery catheter within 10 minutes, putting the animal into hypovolemic shock. This hypovolemic shock state is maintained for 30 minutes. Resuscitation is performed by transfusing 300 microliters per minute (μL / min) of prestored RBCs via the femoral artery until the mean arterial pressure (MAP) stabilizes at 90% of baseline during the 60-minute resuscitation period. MAP and heart rate (HR) are obtained from the femoral artery catheter (PowerLab, AD Instruments, Colorado Springs, CO) at 10, 20, 30, 45, and 60 minutes during this period. After 60 minutes, hematocrit (Hct) is measured by centrifugation of heparinized capillary tubes. Hemoglobin (Hb), lactate, glucose, K+, Na+, pH, and arterial blood gases are determined by an ABL90 co-oximeter (Radiometer, Copenhagen). Indicators of cardiac function and systemic values ​​(MAP, HR, Hct, Hb, and blood gases) are monitored at baseline (BL), during shock, and at 10 (early R), 20, 30, 45, and 60 min (late R) after resuscitation. Animals are euthanized at the end of the experiment. EXAMPLES

[0145] Hematocrit analysis in a rat model of hemorrhagic shock After hypovolemic shock is induced, hematocrit (Hct) is reduced by approximately 30-40%. By providing conventional blood, OR blood, or OCR blood stored for 1 week, hematocrit can be restored to normal levels. See Figure 4A. However, as shown in Figure 4B, 10 minutes after resuscitation (Early R), OR blood stored for 1 week exhibits increased hematocrit percent compared to control and OCR blood. The hematocrit percent of OR blood remains improved compared to control even 60 minutes after resuscitation (Late R). EXAMPLES

[0146] Changes in mean arterial pressure due to deoxygenated blood Mean arterial pressure (MAP) is obtained from a femoral artery catheter (PowerLab, AD Instruments, Colorado Springs, CO). As shown in Figure 5A, the baseline MAP is 80-110 mmHg. MAP is reduced to 20-60 mmHg during hemorrhagic shock. Resuscitation of animals with OR and OCR blood stored for 1 week increases MAP to approximately 80 mmHg and 90 mmHg, respectively. As shown in Figure 5B, resuscitation with OR blood can restore MAP to the normal range compared to control after 10 min. Control and OCR stored blood can restore MAP to the normal range after 60 min of resuscitation. The amount of blood required to perform resuscitation and hemodynamic preservation with conventionally stored RBCs (control) was greater than the OR and OCR RBCs required. See Figure 6A and Figure 6B. EXAMPLES

[0147] Metabolic response to hemorrhagic shock Hemorrhagic shock in animals increases lactate levels from about 2 mmol / L to about 8-14 mmol / L. Resuscitation with OR and OCR RBCs stored for 1 week reduces lactate levels to near normal levels after only 10 minutes of resuscitation. See Figure 7A. Lactate levels in animals resuscitated with control blood are similar to those in animals with hemorrhagic shock. Animals treated with control, OR, and OCR RBCs for 60 minutes show similar lactate levels. As shown in Figure 7B, OR RBCs stored for 3 weeks can also reduce lactate levels compared to controls after 10 minutes of resuscitation. However, after 80 minutes of resuscitation, OCR RBCs restored lactate levels to the normal range. Control and OR RBCs were able to reduce lactate levels, but not to the normal range of 1-3 mmol / L. Analysis of glucose levels shows that the normal range of glucose, about 160 mg / dL to about 240 mg / dL, increases to a range of about 320 to about 510 mg / dL in animals under hemorrhagic shock. See Figure 8A and Figure 8B. Both OR and OCR RBCs stored for 1 week reduce glucose levels compared to the control 10 minutes after resuscitation. All three samples restored glucose levels to the normal range 60 minutes after resuscitation. As shown in Figure 8B, OR and OCR RBCs stored for 3 weeks are also able to reduce glucose levels compared to the control 10 minutes after resuscitation. Unlike RBCs stored for 1 week, only OR and OCR RBCs were able to restore glucose to the normal range. Thus, both lactate and glucose levels are reduced faster in OR and OCR RBCs compared to control RBCs. EXAMPLES

[0148] Organ Injury and Inflammation Animals are analyzed for organ injury and inflammation after experiencing hemorrhagic shock and resuscitation. Elevated liver enzyme levels are indicative of some form of liver damage or injury. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were analyzed to determine liver injury. Resuscitation with OR and OCR RBCs previously stored for 1 week (Figures 9A and 10A) and 3 weeks (Figures 9B and 10B) reduced AST and ALT levels compared to control RBCs stored for the same period. Serum creatinine and blood urea nitrogen (BUN) levels were analyzed to determine renal function. OR and OCR RBCs stored for 1 week reduced serum creatinine levels by more than 30% compared to control RBCs (Figure 11A). After 3 weeks of storage, serum creatinine levels of animals treated with control, OR and OCR RBCs overlap (Figure 11B). BUN levels are reduced by more than 30% in animals treated with 1 week stored OCR RBCs compared to controls (FIG. 12A). BUN levels were also reduced by more than 30% in animals treated with 3 week stored OR RBCs compared to controls (FIG. 12B). Overall, vital organ function was preserved by OR and OCR RBCs compared to control RBCs.

[0149] Upon completion of the in vivo study, the liver, spleen, and lungs are excised from the animals and analyzed for various inflammatory factors, including CXCL1, urinary neutrophil gelatinase-associated lipocalin (u-NGAL), IL-6, and neutrophils. CXCL1 is reduced in animals treated with OR and OCR RBCs stored for 1 or 3 weeks compared to controls stored for the same period (Figure 13A, Figure 13B, Figure 14A, Figure 14B, and Figure 15A, Figure 15B). As shown in Figure 16A and Figure 16B, u-NGAL is reduced in the kidneys of animals treated with OR or OCR RBCs stored for 1 or 3 weeks compared to control RBCs stored for an equivalent period (Figure 16). As shown in Figure 17 and Figure 18, the percentage of lungs excised from animals with CD45+ neutrophils and the levels of IL-6 are significantly reduced in OR and OCR RBCs compared to control RBCs stored for the same period. These results indicate that organ injury and inflammation is reduced in animals treated with OR and OCR RBCs compared to animals treated with control RBCs. EXAMPLES

[0150] Thrombospondin adhesion assay Thrombospondin is an important biomarker of disease severity in sickle cell disease. During vaso-occlusive crises (episodes) in sickle cell disease, levels of circulating thrombospondin are significantly increased with a corresponding increase in adhesion of sickle red blood cells to vascular endothelium. Conventionally stored and oxygen-reduced RBCs are compared to determine differences in adhesion to endothelial cells. Briefly, two units of O positive whole blood are pooled in citrate-phosphate-dextrose (CPD) / SAGM. Each unit is filtered at room temperature through a leukoreduction filter. Leuko-reduced whole blood (LR-WB) is transferred to a 2 liter blood collection bag. The pooled LR-WB is then split into equal aliquots of 500 ml (unit A and unit B) and processed into packed red blood cells and plasma. The plasma is removed and stored at -70°C. Packed RBCs (pRBCs) in unit A are resuspended in SAGM and pRBCs in unit B are resuspended in PAGGSM. The pRBCs in unit B are deoxygenated for 3 hours at room temperature using the Hemanext Oxygen Reduction System. The units are stored at 4°C for 42 days. Samples are extracted from each unit and analyzed to determine complete blood count (CBC), gas panel analysis, and adhesion to thrombospondin on the microchip after preincubation in normal plasma or plasma from sickle cell patients. Samples collected on day 0 are incubated with 0.4% bovine serum albumin (BSA), normal plasma (AA), plasma from sickle cell patients at steady state (SS), or plasma from sickle cell patients in crisis. The incubated samples are then incubated with the adhesion assay microchip. To determine the effect of different shear stresses on the adhesion of oxygen-reduced and conventionally stored RBCs, cells were exposed to 0.5–5 dynes / cm 2The system is subjected to increasing shear stresses of 100 μg / ml. As shown in Figure 19, the system is stable and responsive to changes in shear stress. As the shear stress increases, there is a gradual decrease in the number of cells adhering to the thrombospondin matrix. These results further demonstrate the adhesive strength of RBCs to thrombospondin.

[0151] To test the effect of RBC storage conditions on thrombospondin adhesion, samples collected on days 0, 21, and 42 were incubated at 0.5 dynes / cm 2 (Figure 20A), 2.0 dynes / cm 2 (Figure 20B), or 5.0 dynes / cm 2 (FIG. 20C) As shown in FIG. 20, oxygen-reduced red blood cells are significantly less adhesive to thrombospondin compared to conventionally stored red blood cells. EXAMPLES

[0152] Hemolysis of oxygen-reduced and control RBCs To determine the effect of sickle cell plasma on hemolysis, collect 20 mL samples from two sickle cell patients (TF) and one healthy patient (HT) >28 days after exchange transfusion. Distribute samples into 15 mL centrifuge tubes and centrifuge at 1500 x g for 15 min at room temperature (no brake). Then transfer platelet rich plasma (PRP) from each 15 mL tube to a new 15 mL tube and centrifuge at 1500 x g for 15 min at room temperature (no brake). Using a clean disposable bulb pipette, transfer PRP from each centrifuge tube to a 15 mL pool tube. Dispense 1 mL of platelet poor plasma (PPP) into autoclaved and pre-labeled 1.5 mL tubes. Store PPP at -40 °C until use. Perform flow adhesion with resuspended RBCs in the two TF samples. A TF PPP sample (TF006) is selected that supports a higher baseline adhesion (Figure 21).

[0153] Three sets of pRBCs are prepared and stored: conventionally stored (A), carbon dioxide depleted and 40% oxygen (B), and carbon dioxide and oxygen depleted (C). Aliquots are collected from each set on days 1 and 21 of storage. Samples are transferred to 15 mL centrifuge tubes and centrifuged at 1500×g for 15 minutes. The supernatant is discarded and the RBC pellet is reconstituted. Eighty microliters of concentrated and washed RBCs from A, B, and C are resuspended in 40 microliters of prepared PPP from sickle cell plasma (SS) and healthy plasma (HT) and incubated at 37C for 2 and 24 hours on a nutator (4 RPM). Percent hemolysis is reduced in carbon dioxide and oxygen depleted RBCs compared to conventionally stored RBCs. EXAMPLES

[0154] Effect of sickle plasma on deformability 3 RBC units are pooled, leukodepleted (LR), and equally divided into group A (control) and group B (oxygen reduced). Group B RBCs are processed to contain approximately 5% SO and approximately 8 mmHg CO2. Group A and B RBCs are stored at 42°C for 42 days. Aliquots of RBCs from both groups A and B are incubated at 37°C for (1) 3 hours with healthy donor plasma, (2) 12 hours with healthy donor plasma, (3) 3 hours with sickle cell donor plasma, and (4) 12 hours with sickle cell donor plasma. Matched samples from group A and group B are analyzed side-by-side on a microvascular analysis chip. Deformability is measured as the outlet bulk flow. Deformability is increased in oxygen reduced RBCs incubated with sickle cell plasma compared to control RBCs incubated with healthy plasma. This increase in deformability is observed after 3 and 12 hours of incubation.

Claims

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

2. 10. The method of claim 1, further comprising reducing carbon dioxide in the oxygen-depleted blood.

3. The method of claim 1 , wherein the inflammation is due to trauma, infection, cancer, coagulation disorder, or autoimmunity.

4. 10. The method of claim 1, wherein the patient further has a fever or is at risk of developing a fever.

5. 10. The method of claim 1, wherein the patient has an elevated C-reactive protein (CRP) level.

6. 2. The method of claim 1, wherein the patient has increased 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) compared to patients without fever.

7. 2. The method of claim 1, wherein the patient has reduced levels of one or more inflammatory mediators selected from the group consisting of CCL5 and CXCL10 compared to patients without fever.

8. 10. The method of claim 1, wherein the patient has liver injury, lung injury, spleen injury, kidney injury, bone injury, or a combination thereof.

9. 10. The method of claim 1, wherein the treatment reduces adhesion of red blood cells to thrombospondin in the patient compared to patients treated with conventionally stored blood.

10. 1. A method of improving blood transfusions in a sickle cell patient in need thereof, comprising providing stored oxygen-reduced blood to a patient having sickle cell disease, the oxygen-reduced blood having an oxygen saturation of 20% or less during storage.

11. 11. The method of claim 10, wherein the improvement comprises reducing the incidence of dactylitis (hand-foot syndrome) by at least 10% compared to patients receiving conventionally stored blood.

12. 11. The method of claim 10, wherein the improvement comprises reducing the number of pain crises by at least 10% compared to being treated with conventionally stored blood.

13. 11. The method of claim 10, wherein the improvement comprises reducing complications of anemia selected from the group consisting of fatigue, irritability, dizziness, difficulty breathing, pale skin color, jaundice, growth retardation, and delayed puberty.

14. The method of claim 10 , wherein the improvement comprises reducing the incidence of infection.

15. The method of claim 10, wherein the improvement comprises reducing splenic damage.

16. 11. The method of claim 10, wherein the improvement comprises reducing or preventing stroke.

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

18. 20. The method of claim 17, further comprising reducing carbon dioxide in the oxygen-depleted blood.

19. 18. The method of claim 17, wherein the patient in need thereof has sickle cell disease, inflammation, or sickle cell disease and inflammation.

20. 1. A method for reducing adhesion of red blood cells to endothelial cells in a patient in need thereof, comprising providing stored oxygen-reduced blood to a patient having sickle cell disease, the oxygen-reduced blood having an oxygen saturation of 20% or less during storage.

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