U.S. patent application number 17/241648 was filed with the patent office on 2021-08-26 for methods for the storage of whole blood, and compositions thereof.
This patent application is currently assigned to Hemanext Inc.. The applicant listed for this patent is Hemanext Inc.. Invention is credited to Tatsuro YOSHIDA.
Application Number | 20210260126 17/241648 |
Document ID | / |
Family ID | 1000005567693 |
Filed Date | 2021-08-26 |
United States Patent
Application |
20210260126 |
Kind Code |
A1 |
YOSHIDA; Tatsuro |
August 26, 2021 |
Methods for the Storage of Whole Blood, and Compositions
Thereof
Abstract
Methods and compositions for improved clinical outcomes for
trauma patients receiving whole blood transfusion. Methods and
compositions for improved clinical outcomes for blood transfusions
for cancer patients are also provided.
Inventors: |
YOSHIDA; Tatsuro; (West
Newton, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Hemanext Inc. |
Lexington |
MA |
US |
|
|
Assignee: |
Hemanext Inc.
Lexington
MA
|
Family ID: |
1000005567693 |
Appl. No.: |
17/241648 |
Filed: |
April 27, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15575249 |
Nov 17, 2017 |
11013771 |
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PCT/US2016/033151 |
May 18, 2016 |
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17241648 |
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62163269 |
May 18, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 35/18 20130101;
A61P 7/00 20180101; A61M 1/0272 20130101; A61P 43/00 20180101; A61K
35/14 20130101 |
International
Class: |
A61K 35/18 20060101
A61K035/18; A61K 35/14 20060101 A61K035/14; A61P 7/00 20060101
A61P007/00; A61P 43/00 20060101 A61P043/00 |
Claims
1.-34. (canceled)
35. A leukoreduced whole blood composition for transfusion to a
trauma patient in need of multiple transfusions comprising: oxygen
and carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), and
having a pre-storage and storage oxygen saturation (SO.sub.2) of
30% or less and a pre-storage and storage partial pressure of
carbon dioxide of less than 60 mmHg, and having a 2,3-DPG level at
15 days that is greater than the initial 2,3-DPG level of said
oxygen and carbon dioxide leukoreduced whole blood (OCR-LRWB).
36. The blood composition of claim 35, wherein the 2,3-DPG level of
said OCR-LRWB is at least 80% or higher than the 2,3-DPG level of
the blood at day zero.
37. The blood composition of claim 35, wherein the 2,3-DPG level of
said OCR-LRWB is at least 5 to 20 DPG .mu.mol/gHb after 15 days of
storage.
38. The blood composition of 35, wherein said pre-storage oxygen
saturation is less than 20%.
39. The blood composition of claim 35, wherein said pre-storage
partial pressure of carbon dioxide is between 1 and 60 mmHg.
40. The blood composition of claim 35, wherein said OCR-LRWB
comprises an anticoagulant solution selected from
citrate-phosphate-dextrose with adenine (CPDA1) or
citrate-phosphate-dextrose (CPD).
41. The blood composition of claim 35, wherein said oxygen and
carbon dioxide leukoreduced whole blood (OCR-LRWB) has reduced
levels of a biologic response modifier (BRM) selected from the
group consisting of a cytokine, a chemokine, an isoprostane, and an
oxidized lipid product relative to non-oxygen and carbon dioxide
leukoreduced whole blood (non-OCR-LRWB).
42. The blood composition of claim 41, wherein a patient transfused
with said oxygen and carbon dioxide leukoreduced whole blood
(OCR-LRWB) has a reduced inflammatory response relative to a
patient transfused with non-OCR-LRWB.
43. The blood composition of claim 41, wherein a patient transfused
with said oxygen and carbon dioxide leukoreduced whole blood
(OCR-LRWB) has a reduced immune modulation relative to a patient
transfused with non-OCR-LRWB.
44. The blood composition of claim 41, wherein a patient transfused
with said oxygen and carbon dioxide leukoreduced whole blood
(OCR-LRWB) has a reduced risk of multiple organ dysfunction
relative to a patient transfused with non-OCR-LRWB.
45. The blood composition of claim 41, wherein a patient transfused
with said oxygen and carbon dioxide leukoreduced whole blood
(OCR-LRWB) has a reduced risk of sepsis relative to a patient
transfused with non-OCR-LRWB.
46. The blood composition of claim 41, wherein a patient transfused
with said oxygen and carbon dioxide leukoreduced whole blood
(OCR-LRWB) has a reduced risk of infection relative to a patient
transfused with non-OCR-LRWB.
47. The blood composition of claim 41, wherein a patient transfused
with said oxygen and carbon dioxide leukoreduced whole blood
(OCR-LRWB) has a reduced risk of mortality relative to a patient
transfused with non-OCR-LRWB.
48. The blood composition of claim 41, wherein a patient transfused
with said oxygen and carbon dioxide leukoreduced whole blood
(OCR-LRWB) has a higher red blood cell (RBC) deformability relative
to a patient transfused with non-OCR-LRWB.
49. The blood composition of claim 41, wherein said blood has
equivalent or better coagulation parameters measured by
thromboelastography (TEG) when compared to non-OCR-LRWB.
50. The blood composition of claim 41, wherein said blood has
equivalent or better coagulation parameters as determined by
PT/PTT, fibrinogen, D-dimer, and thrombin generation assays when
compared to non-OCR-LRWB.
51. The blood composition of any one of claim 41, wherein said
blood has equivalent or better platelet function parameters
measured by platelet aggregometer when compared to
non-OCR-LRWB.
52. The blood composition of any one of claim 41, wherein said
blood has equivalent or better levels of clotting factors including
Factor V, Factor VIII, AT, Protein C, or vWF when compared to
non-OCR-LRWB.
53. The blood composition of claim 41, wherein said blood is safe
for transfusion into a patient in need thereof for at least three
weeks.
54.-88. (canceled)
89. The blood composition of claim 35, wherein said pre-storage
oxygen saturation is less than 10% or less than 5%.
90. The blood composition of claim 39, wherein said pre-storage
partial pressure of carbon dioxide is between 10 and 60 mmHg, 20
and 40 mmHg, or 1 and 20 mmHg.
91. The blood composition of claim 35, wherein said patient in need
of multiple transfusions is a trauma patient, transplant patient,
cardiac surgery patient, obstetrics patient, GI surgery patient,
cancer patient, or orthopedic surgery patient.
Description
FIELD OF THE INVENTION
[0001] The present disclosure relates to methods for improving the
quality of whole blood useful for transfusion to patients.
Anaerobic storage of whole blood provides for reduced levels of
cytokines and improved levels of 2,3-diphosphoglycerate (2,3-DPG)
and adenosine triphosphate (ATP). The improved blood compositions
are useful for blood transfusions to cancer and trauma
patients.
BACKGROUND OF THE INVENTION
[0002] When stored conventionally, stored blood undergoes a steady
deterioration which is associated with various storage lesions
including, among others, hemolysis, hemoglobin degradation, and
reduced ATP and 2,3-DPG concentrations. When transfused into a
patient, the effects of the steady deterioration during storage
manifest, for example, as a reduction in the 24-hour in vivo
recovery. Because of these and other medical sequelae of
transfusion of stored blood, a variety of approaches have been
developed to minimize the effects of storage on blood and to
improve medical outcomes. See, for example, Zimring et al.,
"Established and theoretical factors to consider in assessing the
red cell storage lesion" in Blood, 125:2185-90 (2015).
[0003] A number of approaches have been developed aimed at
minimizing storage lesions and improving transfusion outcomes. One
approach has been the development of additive solutions included
during storage. Examples of this approach include U.S. Pat. No.
4,769,318 to Hamasaki et al. and U.S. Pat. No. 4,880,786 to
Sasakawa el al. which are directed to additive solutions for blood
preservation and activation. For example, Rejuvesol.RTM. (available
from Citra Lab LLC, Braintree, Mass.) is added to blood after cold
storage (i.e., 4.degree. C.) just prior to transfusion or prior to
freezing (i.e., at -80.degree. C. with glycerol) for extended
storage. U.S. Pat. No. 6,447,987 to Hess et al. is directed to
additive solutions for the refrigerated storage of human red blood
cells. An alternative approach is to freeze the blood and prevent
the development of storage lesions. Storage of frozen blood is
known in the art, but such frozen blood has limitations. U.S. Pat.
No. 6,413,713 to Serebrennikov is directed to a method of storing
blood at temperatures below 0.degree. C. See Chaplin et al., "Blood
Cells for Transfusion," Blood, 59: 1118-20 (1982), and Valeri et
al., "The survival, function, and hemolysis of human RBCs stored at
4 degrees C. in additive solution (AS-1, AS-3, or AS-5) for 42 days
and then biochemically modified, frozen, thawed, washed, and stored
at 4 degrees C. in sodium chloride and glucose solution for 24
hours," Transfusion, 40:1341-5 (2000). Another approach relates to
the containers for blood storage as provided by U.S. Pat. No.
4,837,047 to Sato et al.
[0004] One approach that has proven successful in improving blood
quality and extending its utility is through the depletion of
oxygen and storage under anaerobic conditions. U.S. Pat. No.
5,624,794 to Bitensky et al., U.S. Pat. No. 6,162,396 to Bitensky
et al., and U.S. Pat. No. 5,476,764 to Bitensky are directed to the
storage of red blood cells under oxygen-depleted conditions. U.S.
Pat. No. 5,789,151 to Bitensky et al. is directed to blood storage
additive solutions. Among the benefits of storing blood under
oxygen depleted conditions are improved levels of ATP and 2,3-DPG,
and reduced hemolysis. Storing blood under oxygen depleted
conditions can also result in reduced microparticle levels,
reductions in the loss of deformability, reduced lipid and protein
oxidation and higher post transfusion survival when compared to
blood stored under conventional conditions.
[0005] U.S. Pat. No. 6,162,396 to Bitensky et al. (the '396 patent)
discloses anaerobic storage bags for blood storage that comprise an
oxygen impermeable outer layer, a red blood cell (RBC) compatible
inner layer that is permeable to oxygen having an oxygen scrubber
placed between the inner and outer layers.
[0006] While the effects of oxygen depletion on packed red blood
cells has been explored, the effects of oxygen depletion on whole
blood has not been reported. In part, the lack of studies on the
deoxygenation of whole blood may be due to the expectation that
deleterious effects are expected when platelets are deprived of
oxygen. More specifically, given the critical role of platelets in
the coagulation process, there were concerns that decreases in
platelet function would result in coagulopathies and negative
consequences to clinical outcomes.
[0007] Storage of platelets has been extensively studied to
identify the most favorable conditions including temperature, pH,
O.sub.2 and CO.sub.2 concentrations. The result of this work is the
conclusion that for stored platelets to persist in a recipient
after transfusion, platelets require access to oxygen and storage
at room temperature. Murphy and Gardner noted in 1975 that unwanted
morphological changes were associated with reduced oxygen
consumption. See. Murphy et al., "Platelet storage at 22 degrees
C.: role of gas transport across plastic containers in maintenance
of viability," Blood 46(2):209-218 (1975). The authors observed
that increased access to oxygen allows for aerobic metabolism
(oxidative phosphorylation) resulting in a reduced rate of lactate
production. At low PO.sub.2 levels lactic acid production is
increased consistent with the Pasteur effect. Moroff et al. noted
that continuous oxygen consumption is required to maintain the pH
of stored platelets at pH 7. See Moroff et al., "Factors
Influencing Changes in pH during Storage of Platelet Concentrates
at 20-24.degree. C.," Vox Sanguinis 42(1):33-45 (1982). Specially
tailored container systems allow permeability to carbon dioxide as
well as oxygen to prevent a lethal drop in pH. As shown by Kakaiya
el al., "Platelet preservation in large containers," Vox Sanguinis
46(2):111-118 (1984), maintaining platelet quality was the result
of improved gas exchange conditions obtained with increased surface
area available for gas exchange. The importance of maintaining
oxygen levels during platelet storage led to the development of gas
permeable containers and storage of platelets in oxygen enriched
atmospheres. See U.S. Pat. No. 4,455,299, issued Jun. 19, 1984, to
Grode. The importance of oxygen to the viability of stored
platelets was reinforced by the observation that in an oxygen poor
environment, the lactate levels increased 5-8 fold. See Kilkson et
al., "Platelet metabolism during storage of platelet concentrates
at 22 degrees C.," Blood 64(2):406-14 (1984). Wallvik et al.,
"Platelet Concentrates Stored at 22.degree. C. Need Oxygen The
Significance of Plastics in Platelet Preservation," Vox Sanguinis
45(4):303-311 (1983), reported that maintaining oxygen during the
first five days of storage was critical for platelet preservation.
Wallvik and co-workers also showed that the maximum platelet number
that can be successfully stored for five days is predictable based
on the determination of the oxygen diffusion capacity of the
storage bag. See Wallvik et al, "the platelet storage capability of
different plastic containers," Vox Sanguinis 58(1):40-4 (1990). By
providing blood bags with adequate gas exchange properties, pH is
maintained, the loss of ATP and the release of alpha-granular
platelet Factor 4 (PF4) was prevented. Each of the foregoing
references are hereby incorporated in their entireties.
[0008] These findings, among others, led to practice
standardization ensuring the oxygenation of platelets during room
temperature storage to maximize post-transfusion viability. When
platelets are stored at refrigerated temperature, post-transfusion
viability is lost, making such platelets unsuitable for
prophylactic transfusion to oncology patients unable to produce
their own platelets. On the other hand, platelets stored at
refrigerated temperature maintain hemostatic functions when
transfused to recipient. Thus, when giving platelets to patients
suffering from traumatic bleeding, viability is less important than
hemostatic activity. We demonstrate that anaerobic storage of
refrigerated whole blood up to 3 weeks yields hemostatic activity
consistent with refrigerated conventionally stored whole blood,
clearly indicating that hemostatic activity of platelets is
maintained with hypothermic storage, even though they are oxygen
starved.
[0009] Though the depletion of oxygen in whole blood has been
mentioned in the literature, the effects of anaerobic storage of
whole blood has not been disclosed. As discussed above, it is well
established that room temperature storage as well as oxygen is
required during storage for long-term survival of platelets (PLT)
(more than 24 hours) in recipients. However, for hemorrhagic trauma
resuscitation, the long-term survival of PLT is not critical
compared to its hemostatic potential. Recently, it became apparent
that patients transfused with stored or fresh whole blood, as well
as reconstituted whole blood (a mixture of plasma, red blood cells
and platelets), have significantly lower post-trauma mortality. We
recently discovered that cold storage enables anaerobic storage of
PLT and also provides known advantages of anaerobically stored RBCs
observed in packed red blood cells, in the whole blood. More
specifically, while unexpectedly preserving the coagulability
without introducing negative effects, deoxygenated whole blood
provides for improved 2,3,-DPG levels. Over a storage period, the
deformability of RBCs is maintained under deoxygenated
conditions.
[0010] Oxidative damage during storage has been implicated as a
major contributor to packed red blood cell (pRBC) membrane damage,
as suggested by the accumulation of markers of lipid peroxidation,
such as isoprostane. Increasing amounts of cytokines during storage
duration may also play a role in storage lesion development with
potential clinical implications for a negative transfusion
outcome.
[0011] Certain patient populations are more susceptible to storage
lesions than others. Among these more sensitive populations are, as
non-limiting examples, trauma patients and cancer patients.
Associated with the adverse clinical outcomes is the accumulation
of biologic response modifiers (BRMs) that include cytokines that
mediate inflammation, regulate cell growth, regulate angiogenesis
and modulate t-helper cell function. Among these BRMs are
interleukin 17 (IL-17), eotaxin (CCL11), basic FGF (bFGF),
macrophage inflammatory protein 1a (MIP-1a), monocyte chemotactic
protein 1 (MCP-1), platelet-derived growth factor (PDGF), tumor
necrosis factor alpha (TNF-.alpha.), and vascular endothelial
growth factor (VEGF). See Behrens et al., "Accumulation of biologic
response modifiers during red blood cell cold storage," Transfusion
49(Suppl3):10A (2009). It has also been observed that cytokines
accumulate during blood storage and these accumulated cytokines can
be associated with negative outcomes when given perioperatively to
cancer patients. See Benson et al., "Accumulation of Pro-Cancer
Cytokines in the Plasma Fraction of Stored Packed Red Cells," J
Gastrointest Surg. 16:460-468 (2012). There is a need for methods
of blood storage that result in reduced levels of BRMs and
cytokines, thereby improving patient outcomes.
[0012] Traumatic injury accounts for 30% of life years lost in the
US, outpacing cancer (16% of life years) and heart disease (12%).
Trauma is the leading cause of death among 1-46 year old patients.
While death from hemorrhage often occurs within 24 hours of
traumatic injury, early death (within 3-6 hours) due to massive
hemorrhage is preventable with prompt and appropriate care.
[0013] Damage Control Resuscitation (DCR) protocols describe the
concept of using balanced ratios of blood components. DCR is
rapidly becoming a standard for arresting hemorrhage and reversing
shock for rapidly hemorrhaging trauma victims. In the civilian
setting, current blood banking practice does not include whole
blood inventory and thus DCR is conducted with a sequential
transfusion of separated components (RBC, plasma and platelets)
such that blood is `reconstituted` in the recipient. Earlier this
year, a large scale randomized controlled trial (RCT), Pragmatic
Randomized Optimal Platelet and Plasma Ratios (PROPPR), was
completed comparing the efficacy of transfusing `reconstituted
blood` at a 1:1:1 unit ratio (plasma, platelets and RBC) vs. a
1:1:2 ratio to trauma patients with massive transfusion. In major
trauma centers, massive transfusion kits combining pre-packaged
blood products consisting of thawed fresh frozen plasma (FFP),
platelets and RBC at 1:1:1 ratios are now readily available.
[0014] Recent studies suggest that whole blood may be superior when
treating patients with severe bleeding to control hemorrhage and
reverse shock for patients with life-threatening bleeding. The 2015
proceedings of the NHLBI State of the Science in Transfusion
Medicine Symposium prioritized the study of whole blood for
patients with severe bleeding. Likewise, the THOR Network, an
international group focused on damage control resuscitation, has
prioritized the comparison of the efficacy and safety of whole
blood to components for hemorrhagic shock. Since modern blood banks
do not routinely supply whole blood, over 80% of level 1 trauma
centers surveyed attempt to mimic the hemostatic, shock-reversing
properties needed in massive transfusion protocols with plasma,
platelets and red blood cells units at ratios of 1:1:1 to 1:1:2 for
both traumatic and non-traumatic life-threatening bleeding cases.
Logistically providing all three blood components rapidly and
safely is difficult, especially given the need to thaw plasma at
centers where an inventory of thawed plasma is not immediately
available. Recent data also indicates that storage of whole blood
at 4.degree. C. for up to 14 days maintains platelet function and
global hemostatic efficacy that is superior to storage at
22.degree. C.
[0015] In addition to a need for blood banks to provide whole blood
for use in certain patient populations, the ability to conserve
valuable blood resources is important. In particular, blood banks
typically discard whole blood stocks after 2 weeks (even though FDA
regulations allow for longer usable lifetimes), thus failing to
take advantage of a valuable and often scarce resource. Such
ability to maximize the value of the blood resource is particularly
useful for small hospitals that serve as Level III and Level IV
trauma centers where an oxygen depleted hemostatic whole blood
product can be maintained under anaerobic conditions and then
processed for packed red blood cells. The present specification
provides for improved whole blood quality for use in trauma
patients, and further provides for an additional source of packed
red blood cells having improved properties and reduced storage
lesions. The present specification overcomes concerns regarding the
wasting of valuable O-negative RBCS (typically used for whole blood
transfusions). Thus, the anaerobic RBCs may be obtained from the
oxygen reduced whole blood and recycled into oxygen depleted RBC
units suitable for storage for up to six weeks. As provided herein,
deoxygenated packed red blood cells may be obtained from the unused
oxygen depleted whole blood, used for transfusion, or stored for
later use under anaerobic conditions.
SUMMARY OF THE INVENTION
[0016] The present disclosure provides for, and includes, a method
for improving survival of a patient in need of multiple
transfusions comprising providing stored red blood cells that have
been oxygen reduced to a patient in need thereof receiving a
medical procedure.
[0017] The present disclosure provides for, and includes, a method
for improving survival of a cancer patient in need thereof
following a perioperative blood transfusion comprising providing
stored red blood cells that have been oxygen reduced to a cancer
patient in need thereof receiving a surgical procedure.
[0018] The present disclosure provides for, and includes, a method
for reducing pro-cancer cytokines in stored blood comprising
depleting oxygen from the blood prior to storage comprising
collecting blood in an anticoagulant solution, reducing the
leukocytes from the collected blood, reducing the pre-storage
oxygen saturation (SO.sub.2) to 30% or less and a pre-storage
partial pressure of carbon dioxide to less than 60 mmHg; and
storing the oxygen and carbon dioxide reduced blood under anaerobic
conditions.
[0019] The present disclosure provides for, and includes, a blood
composition for transfusion to a trauma patient in need thereof
comprising deoxygenated leukoreduced whole blood in an
anticoagulant solution and having a pre-storage oxygen saturation
(SO.sub.2) of 20% or less and a pre-storage partial pressure of
carbon dioxide of less than 60 mmHg wherein the deoxygenated
leukoreduced whole blood has a 2,3-DPG level at 15 days that is
greater than the initial 2,3-DPG level of the deoxygenated
leukoreduced blood.
[0020] The present disclosure provides for, and includes, a method
of reducing an inflammatory response in a patient receiving a blood
transfusion comprising transfusing an oxygen depleted blood product
to a patient in need thereof, wherein the oxygen depleted whole
blood has reduced levels of inflammatory cytokines after storage
under anaerobic conditions.
[0021] The present disclosure provides for, and includes, a method
of reducing an immune response in a patient receiving a blood
transfusion comprising transfusing an oxygen depleted blood product
to a patient in need thereof, wherein the oxygen depleted blood
product has reduced levels of a cytokine after storage under
anaerobic conditions. In aspects according to the present
disclosure, an immune response is an immune modulation or immune
suppression. In other aspects, the immune response is an
activation, including for example, inflammation.
[0022] The present disclosure provides for, and includes, a method
for improving perfusion of oxygen in a patient in need thereof
comprising transfusing an oxygen depleted blood product to a
patient in need thereof, wherein the oxygen depleted blood product
has higher RBC deformability compared to a conventionally stored
blood product.
[0023] The present disclosure provides for, and includes, a method
for managing a blood bank comprising maintaining an inventory of
blood units comprising oxygen reduced whole blood and an
anticoagulant, or oxygen reduced leukoreduced whole blood and an
anticoagulant; providing one or more of the blood units from the
inventory for treatment of a patient; and recycling blood units
from the inventory to prepare component separated deoxygenated
blood units. The present disclosure further provides for using
recycled blood units to prepare reconstituted blood units for
treatment of trauma patients requiring massive transfusions.
[0024] The present disclosure provides for, and includes, a method
of providing a supply of blood products for transfusion medicine
comprising depleting oxygen or oxygen and carbon dioxide from whole
blood to prepare oxygen or oxygen and carbon dioxide reduced whole
blood; and storing the oxygen or oxygen and carbon dioxide reduced
whole blood for a time period and providing the stored blood to a
patient in need thereof; or storing the oxygen or oxygen and carbon
dioxide reduced whole blood for a time period, and preparing oxygen
or oxygen and carbon dioxide reduced packed red blood cells.
BRIEF DESCRIPTION OF THE DRAWINGS
[0025] The present disclosure is disclosed with reference to the
accompanying drawings, wherein:
[0026] FIGS. 1A to 1D are graphs displaying the results of cytokine
measurements showing reduced levels of eotaxin (1A) and RANTES (1B)
in anaerobically stored packed red blood cells. FIG. 1C shows
reduced levels of cell free hemoglobin compared to aerobically
stored packed red blood cells. FIG. 1D shows reduced levels of
isoprostane in anaerobically stored packed red blood cells. Dashed
lines=aerobically stored blood; Solid lines=anaerobically stored
blood.
[0027] FIGS. 2A to 2G are graphs presenting the results of two
experiments according to the present disclosure comparing the
storage of leukoreduced whole blood collected in anticoagulant
solution CPD (LRWB/CPD) under oxygen reduced, oxygen and carbon
dioxide reduced and conventionally stored LRWB/CPD over a period of
21 days. FIG. 2A presents the levels 2,3-DPG. FIG. 2B presents the
levels of ATP. FIG. 2C presents the pH. FIG. 2D presents the
platelet count. FIG. 2E presents potassium levels. FIG. 2F presents
the data of FIG. 2A re-plotted relative to the levels of 2,3-DPG at
day 0 (T.sub.0). FIG. 2G presents the data of FIG. 2B re-plotted
relative to the levels of ATP at day 0 (T.sub.0). Key: sample
c68/80 is conventionally stored blood having an initial oxygen
saturation of 68% and a partial pressure of CO.sub.2 of 80 mmHg;
sample c50/94 is conventionally stored blood having an initial
oxygen saturation of 50% and a partial pressure of CO.sub.2 of 94
mmHg; sample sc91/75 is conventionally stored blood having an
initial oxygen saturation of 91% and a partial pressure of CO.sub.2
of 75 mmHg; sample sc69/87 is conventionally stored blood having an
initial oxygen saturation of 69% and a partial pressure of CO.sub.2
of 87 mmHg; sample tc5/78 is oxygen depleted, anaerobically stored
blood having an initial oxygen saturation of 5% and a partial
pressure of CO.sub.2 of 78 mmHg; sample tc7/64 is oxygen depleted,
anaerobically stored blood having an initial oxygen saturation of
7% and a partial pressure of CO.sub.2 of 64 mmHg; sample T5/28 is
oxygen and carbon dioxide depleted, anaerobically stored blood
having an initial oxygen saturation of 5% and a partial pressure of
CO.sub.2 of 28 mmHg; sample T4/26 is oxygen and carbon dioxide
depleted, anaerobically stored blood having an initial oxygen
saturation of 4% and a partial pressure of CO.sub.2 of 26 mmHg.
[0028] FIGS. 3A to 3D are graphs presenting the results of two
experiments according to the present disclosure comparing the
storage of leukoreduced whole blood collected in anticoagulant
solution CPDA1 (LRWB/CPDA1) under oxygen reduced, oxygen and carbon
dioxide reduced and conventionally stored LRWB/CPDA1 over a period
of 21 days. FIG. 3A presents the levels 2,3-DPG. FIG. 3B presents
the levels of ATP. FIG. 3C presents the data of FIG. 3A re-plotted
relative to the levels of 2,3-DPG at day 0 (T.sub.0). FIG. 3D
presents the data of FIG. 3B re-plotted relative to the levels of
ATP at day 0 (T.sub.0). Key: sample c32/98 is conventionally stored
blood having an initial oxygen saturation of 32% and a partial
pressure of CO.sub.2 of 98 mmHg; sample c56/86 is conventionally
stored blood having an initial oxygen saturation of 56% and a
partial pressure of CO.sub.2 of 86 mmHg; sample sc59/95 is
conventionally stored blood having an initial oxygen saturation of
59% and a partial pressure of CO.sub.2 of 95 mmHg; sample sc82/84
is conventionally stored blood having an initial oxygen saturation
of 82% and a partial pressure of CO.sub.2 of 84 mmHg; sample tc7/80
is oxygen depleted, anaerobically stored blood having an initial
oxygen saturation of 7% and a partial pressure of CO.sub.2 of 80
mmHg; sample tc6/77 is oxygen depleted, anaerobically stored blood
having an initial oxygen saturation of 6% and a partial pressure of
CO.sub.2 of 77 mmHg; sample T5/28 is oxygen and carbon dioxide
depleted, anaerobically stored blood having an initial oxygen
saturation of 5% and a partial pressure of CO.sub.2 of 28 mmHg;
sample T7/23 is oxygen and carbon dioxide depleted, anaerobically
stored blood having an initial oxygen saturation of 7% and a
partial pressure of CO.sub.2 of 23 mmHg.
[0029] FIGS. 4A to 4C are graphs presenting the results of
experiments according to the present disclosure comparing the
storage of leukoreduced whole blood collected in anticoagulant
solution CPDA1 (LRWB/CPDA1) under oxygen reduced (OR), oxygen and
carbon dioxide reduced (OCR) and conventionally stored LRWB/CPDA1
over a period of 21 days. FIG. 4A presents the levels of ATP in
OR-LRWB/CPDA1, OCR-LRWB/CPDA1 and conventionally stored LRWB/CPDA1.
FIG. 4B presents the levels of 2,3-DPG in OR-LRWB/CPDA1,
OCR-LRWB/CPDA1 and conventionally stored LRWB/CPDA1. FIG. 4C
presents the percent hemolysis in OR-LRWB/CPDA1, OCR-LRWB/CPDA1 and
conventionally stored LRWB/CPDA1. In graphs presented in FIGS. 4A
to 4C, small dashed lines=OR-LRWB/CPDA1 stored blood, dashed
lines=ORC-LRWB/CPDA1 stored blood, and solid lines=conventionally
stored blood.
[0030] FIGS. 5A to 5D are graphs presenting the results of
experiments according to the present disclosure comparing the
storage of leukoreduced whole blood collected in anticoagulant
solution CPDA1 (LRWB/CPDA1) under oxygen and carbon dioxide reduced
and conventionally stored LRWB/CPDA1 over a period of 21 days. FIG.
5A presents activated Partial Thrombin Time in seconds (aPTT) in
OCR-LRWB/CPDA1 and conventionally stored LRWB/CPDA1. FIG. 5B
presents prothrombin time in seconds (PT) in OCR-LRWB/CPDA1 and
conventionally stored LRWB/CPDA1. FIG. 5C presents the levels of
Fibrinogen in OCR-LRWB/CPDA1 and conventionally stored LRWB/CPDA1.
FIG. 5D presents the levels of D-dimer in OCR-LRWB/CPDA1 and
conventionally stored LRWB/CPDA1. In graphs presenting plasma
coagulation parameters, dashed lines=ORC-LRWB/CPDA1 stored blood,
and solid lines=conventionally stored blood.
[0031] FIGS. 6A to 6E are graphs presenting the results of
experiments according to the present disclosure comparing the
storage of leukoreduced whole blood collected in anticoagulant
solution CPDA1 (LRWB/CPDA1) under oxygen and carbon dioxide reduced
and conventionally stored LRWB/CPDA1 over a period of 21 days. FIG.
6A presents levels of Factor V in OCR-LRWB/CPDA1 and conventionally
stored LRWB/CPDA1. FIG. 6B presents levels of Factor VIII in
OCR-LRWB/CPDA1 and conventionally stored LRWB/CPDA1. FIG. 6C
presents Protein C activity in OCR-LRWB/CPDA1 and conventionally
stored LRWB/CPDA1. FIG. 6D presents Protein S activity in
OCR-LRWB/CPDA1 and conventionally stored LRWB/CPDA1. FIG. 6E
presents levels of von Willebrand Factor (vWF) in OCR-LRWB/CPDA1
and conventionally stored LRWB/CPDA1. In graphs presenting plasma
clotting factors, dashed lines=ORC-LRWB/CPDA1 stored blood, and
solid lines=conventionally stored blood.
[0032] FIGS. 7A to 7D are graphs presenting the results of
experiments according to the present disclosure comparing the
storage of leukoreduced whole blood collected in anticoagulant
solution CPDA1 (LRWB/CPDA1) under oxygen and carbon dioxide reduced
(OCR) and conventionally stored LRWB/CPDA1 over a period of 21
days. FIG. 7A presents the speed at which fibrin build up and
crosslinking takes place (TEG Angle) in OCR-LRWB/CPDA1 and
conventionally stored LRWB/CPDA1. FIG. 7B presents a comparison of
blood kinetics (TEG K) in OCR-LRWB/CPDA1 and conventionally stored
LRWB/CPDA1. FIG. 7C presents the maximum amplitude in
OCR-LRWB/CPDA1 and conventionally stored LRWB/CPDA1. FIG. 7D
presents the reaction time in OCR-LRWB/CPDA1 and conventionally
stored LRWB/CPDA1. In graphs presenting thromboelastography (TEG)
parameters, dashed lines=ORC-LRWB/CPDA1 stored blood, and solid
lines=conventionally stored blood.
DETAILED DESCRIPTION
[0033] Unless defined otherwise, technical and scientific terms as
used herein have the same meaning as commonly understood by one of
ordinary skill in the art. One skilled in the art will recognize
many methods can be used in the practice of the present disclosure.
Indeed, the present disclosure is in no way limited to the methods
and materials described. Any references cited herein are
incorporated by reference in their entireties. For purposes of the
present disclosure, the following terms are defined below.
[0034] As used herein, the term "patient" includes a person in need
of a medical procedure receiving a blood product.
[0035] As used herein, the term "multiple transfusion" includes a
patient receiving more than 195 units of blood. In another aspect,
a multiple transfusion can include a patient receiving at least
1.times.10.sup.5 mL of blood. In another aspect, multiple
transfusion includes a patient receiving from 1 to 1.times.10.sup.5
mL of blood. In another aspect, multiple transfusion includes a
patient receiving from 1.times.10.sup.4 to 1.times.10.sup.5 mL of
blood.
[0036] As used herein, the term "blood" refers to whole blood,
leukoreduced RBCs, platelet reduced RBCs, and leukocyte and
platelet reduced RBCs. The term blood further includes packed red
blood cells, platelet reduced packed red blood cells, leukocyte
reduced packed red blood cells (LRpRBC), and leukocyte and platelet
reduced packed red blood cells. The temperature of blood can vary
depending on the stage of the collection process, starting at the
normal body temperature of 37.degree. C. at the time and point of
collection, but decreasing rapidly to about 30.degree. C. as soon
as the blood leaves the patient's body and further thereafter to
room temperature in about 6 hours when untreated, and ultimately
being refrigerated at between about 4.degree. C. and 6.degree.
C.
[0037] As used herein, "blood product" includes separated
platelets, plasma, or white blood cells.
[0038] As used herein, "recovered blood product" includes separated
platelets, plasma, or white blood cells collected from a donor.
[0039] As used herein. "recovered blood" includes whole blood and
red blood cells collected from a donor and previously stored under
oxygen reduced conditions. In one aspect of the present disclosure,
suitable blood for use in this method includes oxygen reduced
leukoreduced packed red blood cells (OR-LRpRBC), oxygen reduced
leukoreduced packed red blood cells with platelets (OR-LRpRBC+PLT),
oxygen and carbon dioxide reduced leukoreduced packed red blood
cells (OCR-LRpRBC), or oxygen and carbon dioxide reduced
leukoreduced packed red blood cells with platelets (OCR-LRpRBC+PLT)
obtained from oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) after at least one week
of storage. In another aspect, the suitable blood for use in this
method is stored for up to 42 days. In another aspect, the suitable
blood for use in this method is stored for up to 56 days. In
another aspect, the suitable blood for use in this method is stored
for up to 64 days.
[0040] As used herein, a method of obtaining "component separated
blood products" includes obtaining recycled blood from a blood bank
inventory and separating into platelets, plasma, and white blood
cells. Suitable blood for use in this method comprises oxygen
reduced whole blood having an anticoagulant and oxygen reduced
leukoreduced whole blood having an anticoagulant. In an aspect of
the present disclosure, a component separated oxygen reduced blood
is stored for up to six weeks. In another aspect, a component
separated oxygen reduced blood includes an additive solution. In
certain aspects, the additive solution may be AS-1. In certain
aspects, the additive solution is AS-3 (Nutricel.RTM.). In certain
aspects, the additive solution is AS-5. In certain aspects, the
additive solution is SAGM. In certain aspects, the additive
solution is PAGG-SM. In certain aspects, the additive solution is
PAGG-GM. In certain aspects, the additive solution is MAP. In
certain aspects, the additive solution is SOLX. In certain aspects,
the additive solution is ESOL. In certain aspects, the additive
solution is EAS61. In certain aspects, the additive solution is
OFAS1. In certain aspects, the additive solution is OFAS3. In
certain aspects, the additive solution is a combination of AS-1,
AS-3 (Nutricel.RTM.), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, SOLX,
ESOL, EAS61, OFAS1, and OFAS3, alone or in combination.
[0041] As used herein, "reconstituted WB" includes providing
platelets. RBC, and plasma in parallel to a patient during
transfusion.
[0042] As used herein, "derived WB" includes oxygen reduced and
oxygen and carbon dioxide reduced whole blood.
[0043] As used herein, "stored red blood cells" includes oxygen
reduced or oxygen and carbon dioxide reduced red blood cells stored
from 1 to 6.degree. C. In an aspect, stored red blood cells include
red blood cells (RBC) present in whole blood. In another aspect,
stored red blood cells include RBC present in leukoreduced whole
blood. In another aspect, stored red blood cells include red blood
cells (RBC) present in leukoreduced RBC. In a further aspect,
stored red blood cells include red blood cells (RBC) present in
platelet reduced RBC. In yet another aspect, stored red blood cells
include red blood cells (RBC) present in leukoreduced and platelet
reduced RBC.
[0044] As used herein, "whole blood" includes white blood cells
(WBCs), platelets suspended in plasma, and includes electrolytes,
hormones, vitamins, antibodies, etc. In whole blood, white blood
cells are normally present in the range of between 4.5 and
11.0.times.10.sup.9 cells/L, and the normal RBC range at sea level
is 4.6-6.2.times.10.sup.12/L for men and 4.2-5.4.times.10.sup.12/L
for women. The normal hematocrit, or percent packed cell volume, is
about 40-54% for men and about 38-47% for women. The platelet count
is normally 150-450.times.10.sup.9/L for both men and women. Whole
blood is collected from a blood donor, and is usually combined with
an anticoagulant. Whole blood, when collected is initially at about
37.degree. C. and rapidly cools to about 30.degree. C. during and
shortly after collection, but slowly cools to ambient temperature
over about 6 hours. Whole blood may be processed according to
methods of the present disclosure at collection, beginning at
30-37.degree. C., or at room temperature (typically about
25.degree. C.). As used herein, a "unit" of blood is about 450-500
ml including anticoagulant. Suitable anticoagulants include CPD,
CPDA1, ACD, and ACD-A. As used herein, "time collected" (Tc) is the
time at which blood is collected from the patient.
[0045] As used herein, "red blood cells" (RBCs), stored red blood
cells, oxygen reduced red blood cells, and oxygen and carbon
dioxide reduced red blood cells, include RBCs present in whole
blood, leukoreduced RBCs, platelet reduced RBCs, leukocyte and
platelet reduced RBCs, and packed red blood cells (pRBCs). Human
red blood cells in vivo are in a dynamic state. The red blood cells
contain hemoglobin, the iron-containing protein that carries oxygen
throughout the body and gives red blood its color. The percentage
of blood volume composed of red blood cells is called the
hematocrit. As used herein, unless otherwise limited, RBCs also
includes 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 pRBCs is about 70%. As used herein, oxygen reduced
RBC (OR-RBC) can include oxygen and carbon dioxide (OCR-) reduced
RBC (OCR-RBC)
[0046] As used herein, "leukoreduced whole blood" (LRWB) includes
whole blood having an anticoagulant that has been treated to remove
white blood cells and platelets, usually by filtration or
centrifugation. Leukoreduced whole blood has levels of white blood
cells that are reduced by at least 5 logs.
[0047] As used herein, "oxygen reduced leukoreduced whole blood"
(OR-LRWB) can include oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB).
[0048] As used herein, "leukoreduced whole blood with platelets"
(LRWB+PLT) includes oxygen reduced (OR-) whole blood having an
anticoagulant and leukoreduced with a platelet-sparing filter. As
used herein, oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT) can include oxygen and carbon dioxide reduced
leukoreduced whole blood with platelets (OCR-LRWB+PLT).
[0049] As used herein, "leukoreduced packed red blood cells"
(LRpRBC) includes packed red blood cells having oxygen reduced
(OR-) whole blood having an anticoagulant that has been treated to
remove white blood cells, usually by filtration or centrifugation.
As used herein, oxygen reduced leukoreduced packed red blood cells
(OR-LRpRBC) can include oxygen and carbon dioxide reduced
leukoreduced packed red blood cells (OCR-LRpRBC).
[0050] As used herein, "leukoreduced packed red blood cells with
platelets" (LRpRBC+PLT) includes packed red blood cells having
platelets obtained from oxygen reduced whole blood having an
anticoagulant that has been treated to remove white blood cells
with a platelet-sparing filter. As used herein, oxygen reduced
leukoreduced packed red blood cells with platelets (OR-LRpRBC+PLT)
can include oxygen and carbon dioxide reduced leukoreduced packed
red blood cells with platelets (OCR-LRpRBC+PLT).
[0051] In aspects of the present disclosure, the method and
compositions may include adding an additive solution to the packed
RBCs to form a suspension. A number of additive solutions are known
in the art. In certain aspects, the additive solution may be
selected from the group consisting of AS-1, AS-3 (Nutricel.RTM.),
AS-5, SAGM. PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, and
OFAS3, alone or in combination. Additive AS-1 is disclosed in
Heaton el al., "Use of Adsol preservation solution for prolonged
storage of low viscosity AS-1 red blood cells," Br J Haematol.,
57(3):467-78 (1984). In a further aspect, the additive solution may
have a pH of from 5.0 to 9.0. In another aspect, the additive may
include an antioxidant. In some aspects according the present
disclosure, the antioxidant may be quercetin, alpha-tocopherol,
ascorbic acid, or enzyme inhibitors for oxidases.
[0052] As used herein the term "about" refers to .+-.10%.
[0053] The terms "comprises," "comprising," "includes,"
"including," "having," and their conjugates mean "including but not
limited to."
[0054] The term "consisting of" means "including and limited
to."
[0055] The term "consisting essentially of" means that the
composition, method or structure may include additional
ingredients, steps and/or parts, but only if the additional
ingredients, steps and/or parts do not materially alter the basic
and novel characteristics of the claimed composition, method or
structure.
[0056] 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" may include a plurality of compounds, including mixtures
thereof.
[0057] Throughout this application, various aspects of this
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. Accordingly,
the description of a range should be considered to have
specifically disclosed all the possible subranges as well as
individual numerical values within that range. For example,
description of a range such as "from 1 to 6" should be considered
to have specifically disclosed subranges such as "from 1 to 3,"
"from 1 to 4," "from 1 to 5," "from 2 to 4," "from 2 to 6," "from 3
to 6," etc., as well as individual numbers within that range, for
example, 1, 2, 3, 4, 5, and 6. This applies regardless of the
breadth of the range.
[0058] Whenever a numerical range is indicated herein, it is meant
to include any cited numeral (fractional or integral) within the
indicated range. The phrases "ranging/ranges between" a first
indicate number and a second indicate number and "ranging/ranges
from" a first indicate number "to" a second indicate number are
used herein interchangeably and are meant to include the first and
second indicated numbers and all the fractional and integral
numerals therebetween.
[0059] As used herein the term "method" refers to manners, means,
techniques, and procedures for accomplishing a given task
including, but not limited to, those manners, means, techniques,
and procedures either known to or readily developed from known
manners, means, techniques, and procedures by practitioners of the
chemical, pharmacological, biological, biochemical, and medical
arts.
[0060] As used herein, the term "equivalent" means that the
measured values of oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT), when compared the
measured values of otherwise equivalently treated conventionally
stored blood, are within 1 standard deviation of each other with a
sample size of at least 5 for each compared measured condition.
[0061] As used herein, the term "greater" or "increased" means that
the measured values of oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT), when comparedOR-WB, when
compared to the measured values of otherwise equivalently treated
conventionally stored blood, are at least 1 standard deviation
greater, with a sample size of at least 5 for each compared
measured condition.
[0062] As used herein, the term "decreased" or "less" means that
the measured values of oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT), when comparedOR-WB, when
compared to the measured values of otherwise equivalently treated
conventionally stored blood, are at least 1 standard deviation
lower, with a sample size of at least 5 for each compared measured
condition.
[0063] As used herein, the terms "conventionally stored,"
"conventional storage," and "conventional conditions" include whole
blood, leukoreduced RBCs, platelet reduced RBCs, leukocyte and
platelet reduced RBCs, packed red blood cells, platelet reduced
packed red blood cells, leukocyte reduced packed red blood cells
(LRpRBC), and leukocyte and platelet reduced packed red blood cells
stored in oxygen and carbon dioxide permeable container at 1 to
6.degree. C. without gas reduction steps prior to storage. In an
aspect of the present disclosure, both oxygen and carbon dioxide
increase to ambient levels over time in conventionally stored whole
blood, leukoreduced RBCs, platelet reduced RBCs, leukocyte and
platelet reduced RBCs, packed red blood cells, platelet reduced
packed red blood cells, leukocyte reduced packed red blood cells
(LRpRBC), and leukocyte and platelet reduced packed red blood
cells, due to oxygen and carbon dioxide container permeability.
While not traditionally considered conventional, for the purposes
of the present disclosure, conventional storage can include storage
at temperatures above 6.degree. C. Also, while not traditionally
considered conventional, for the purposes of the present
disclosure, conventional storage can include storage at freezing
temperatures.
[0064] The present disclosure provides for, and includes, methods
to provide desirable characteristics to blood products for
transfusion. It has been discovered that depletion of oxygen from
packed red blood cells results in reduced accumulations of unbound
cytokine, particularly RANTES (C--C motif chemokine ligand 5, CCL5)
and eotaxin (C--C motif chemokine ligand 11, CCL11), as well as
cell-free hemoglobin, and 8-isoprostane F.sub.2a. Not to be limited
by theory, it is thought that RANTES and eotaxin are normally
sequestered by binding to the DARC (atypical chemokine receptor 1,
ACKR1) and oxidative stress damages DARC and releases the bound
chemokines. Thus, while overall content of the chemokines does not
change, the effective concentration (e.g., freely diffusible and
unbound) increases and is then available to affect a transfused
patient. As will be understood, the presence of these active
chemokines (acting in a dose dependent manner) can be detrimental
to trauma and other patients receiving two or more transfusions.
These findings demonstrate an unexpected benefit of anaerobically
stored blood, in addition to the desirable efficient oxygen
delivery associated with elevated 2,3-DPG values, and provides a
potential reduction in some of the components of the storage lesion
resultant from pRBC oxidative damage during storage. These
cytokines are known to be negatively associated with patient
outcome in some patient populations. Accordingly, the discovery
that unbound cytokine accumulation can be reduced provides for
improved methods of treating patients susceptible to cytokines.
[0065] The present disclosure provides for, and includes, improving
the survival of a patient in need of multiple transfusions by
providing stored red blood cells that have been oxygen reduced
(OR-stored RBCs) to a patient in need thereof receiving a medical
procedure. Not to be limited by theory, it is believed that
increased levels of cytokines have adverse effects on recipient
patients that increases morbidity. In an aspect, the stored red
blood cells are oxygen reduced (OR). In a further aspect, the
stored red blood cells are both oxygen and carbon dioxide reduced
(OCR). As shown in the examples, in OCR samples, the levels of ATP
are decreased and maintained at lower levels for at least 15 days,
while in OR samples, the levels of ATP are increased compared to
conventionally stored samples (see FIG. 4A). As shown in the
examples, in OCR samples, the levels of 2,3-DPG are increased and
maintained at high levels for at least 15 days, while in OR
samples, 2,3-DPG levels are increased over conventional storing but
not as high as 2,3-DPG levels of OCR samples (see FIG. 4B).
Further, as shown in the examples, hemolysis is equivalent in OR,
OCR, and conventionally stored samples.
[0066] In an aspect of the present disclosure, cytokines comprise
monocyte chemotactic protein-1 (MCP-1). In another aspect,
cytokines comprise regulated on activation normal T cell expressed
and secreted (RANTES). In another aspect, cytokines comprise
angiogenin. In another aspect of the present disclosure, cytokines
comprise tumor necrosis factor-alpha (TNF-.alpha.). In another
aspect, cytokines comprise epidermal growth factor (EGF). In a
further aspect, cytokines comprise platelet-derived growth factor
(PDGF).
[0067] In an aspect of the present disclosure, the level of the
factor RANTES is less than 500 pg/ml after 21 days under OR
conditions. In another aspect, the level of the factor RANTES is
less than 400 pg/ml after 21 days under OR conditions. In another
aspect, the level of the factor RANTES is less than 300 pg/ml after
21 days under OR conditions. In another aspect, the level of the
factor RANTES is more than 100 pg/ml after 21 days under OR
conditions. In a further aspect, the level of the factor RANTES is
from 0 to 300 pg/ml after 21 days under OR conditions.
[0068] In an aspect of the present disclosure, the level of the
factor eotaxin is less than 150 pg/ml after 21 days under OR
conditions. In another aspect, the level of the factor eotaxin is
less than 100 pg/ml after 21 days under OR conditions. In another
aspect, the level of the factor eotaxin is from 0 to 100 pg/ml
after 21 days under OR conditions. In another aspect, the level of
the factor eotaxin is preferably 100 pg/ml after 21 days under OR
conditions. In another aspect, the level of the factor eotaxin is
more than 100 pg/ml after 21 days under OR conditions. In a further
aspect, the level of the factor eotaxin is from 0 to 300 pg/ml
after 21 days under OR conditions.
[0069] In aspects according to the present disclosure, the
OR-stored RBCs are selected from the group consisting of oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen
reduced leukoreduced packed red blood cells (OR-LRpRBC), oxygen
reduced leukoreduced packed red blood cells with platelets
(OR-LRpRBC+PLT), oxygen and carbon dioxide reduced leukoreduced
whole blood (OCR-LRWB), oxygen and carbon dioxide reduced
leukoreduced whole blood with platelets (OCR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced packed red blood cells
(OCR-LRpRBC), oxygen and carbon dioxide reduced leukoreduced packed
red blood cells with platelets (OCR-LRpRBC+PLT), and combinations
thereof. In other aspects, wherein said OR-stored RBCs comprise
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT).
[0070] In an aspect, a patient in need of multiple transfusions is
a trauma patient. In another aspect, a patient in need of multiple
transfusions is a transplant patient. In another aspect, a patient
in need of multiple transfusions is a cardiac surgery patient. In
another aspect, a patient in need of multiple transfusions is an
obstetrics patient. In another aspect, a patient in need of
multiple transfusions is a gastrointestinal (GI) surgery patient.
In a further aspect, a patient is an orthopedic surgery
patient.
[0071] In an aspect, a patient in need of multiple transfusions is
a trauma patient. In another aspect, a patient in need of multiple
transfusions is a hemorrhagic trauma patient. In a further aspect,
a patient in need of multiple transfusions is a blunt trauma
patient.
[0072] In an aspect, the reduction of cytokines in oxygen reduced
stored packed red blood cells provides for improved treatment of
cancer patients in need of blood transfusions. It is known in the
art that cytokines are associated with negative patient outcome for
patients receiving perioperative blood transfusions for surgical
treatments of cancer patients. In an aspect, the oxygen depleted,
cytokine reduced blood products are provided to a cancer patient
prior to undergoing surgery. In another aspect, the oxygen
depleted, cytokine reduced blood products are provided to a cancer
patient during surgery. In another aspect, the oxygen depleted,
cytokine reduced blood products are provided to a cancer patient
following surgery.
[0073] In aspects according to the present disclosure, the oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) has higher levels of 2,3-DPG compared to
conventionally stored leukoreduced whole blood (WB) and provides
for improved oxygen delivery. Under anaerobic conditions, 2,3-DPG
levels can be maintained in whole blood for up to 4 weeks. In an
aspect, the 2,3-DPG levels are maintained above 50% of physiologic
levels for up to four weeks. In aspects according to the present
disclosure, improved 2,3-DPG levels are maintained for at 2 weeks.
In other aspects, 2,3-DPG levels are maintained for three weeks. In
an aspect, the 2,3-DPG level of the oxygen reduced leukoreduced
whole blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) is
at least 80% or higher than the 2,3-DPG level of the blood at day
zero. In another aspect, the 2,3-DPG level of the oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) is at least 5 to 20 DPG .mu.mol/gHb.
[0074] Also provided for and included in the present disclosure is
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that have depleted whole blood (OR-WB)
that has reduced levels of biological response modifiers (BRMs)
relative to conventionally stored whole blood. In certain aspects,
the BRM present in oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is about half the level
of conventionally stored blood after 21 days. In an aspect, the
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) has levels of cytokines that are
relatively unchanged after 10 days in storage under anaerobic
conditions. In another aspect, the cytokine level is relatively
unchanged after 30 days of storage. In another aspect, the cytokine
level is relatively unchanged after 40 days of storage. As used
herein, "relatively unchanged" means that the concentration of
cytokine, normalized to hemoglobin levels is within 1 standard
deviation of the initial normalized concentration of cytokine.
[0075] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of the cytokine eotaxin compared to conventionally
stored whole blood. In an aspect, the level of eotaxin in oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is about half the level of eotaxin present
in conventionally stored blood after 21 days, normalized to the
hemoglobin concentration. In an aspect, the level of eotaxin in
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is about 25% or less of the level of
eotaxin present in conventionally stored blood after 40 days.
[0076] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of the cytokine RANTES (regulated on activation,
normal T cell expressed and secreted) compared to conventionally
stored whole blood. In an aspect, the level of RANTES in oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is about half the level of RANTES present
in conventionally stored blood after 21 days, normalized to the
hemoglobin concentration. In an aspect, the level of RANTES in
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is about 25% or less of the level of
RANTES present in conventionally stored blood after 40 days.
[0077] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of monocyte chemotactic protein-1 (MCP-1) compared
to conventionally stored whole blood. In an aspect, the level of
MCP-1 in oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) is about half the level of
MCP-1 present in conventionally stored blood after 21 days,
normalized to the hemoglobin concentration. In an aspect, the level
of MCP-1 in oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is about 25% or less of
the level of MCP-1 present in conventionally stored blood after 40
days.
[0078] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of angiogenin compared to conventionally stored
whole blood. In an aspect, the level of angiogenin in oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is about half the level of angiogenin
present in conventionally stored blood after 21 days, normalized to
the hemoglobin concentration. In an aspect, the level of angiogenin
in oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) is about 25% or less of the
level of angiogenin present in conventionally stored blood after 40
days.
[0079] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of tumor necrosis factor-alpha (TNF-.alpha.)
compared to conventionally stored whole blood. In an aspect, the
level of TNF-.alpha. in oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is about half the level
of TNF-.alpha. present in conventionally stored blood after 21
days, normalized to the hemoglobin concentration. In an aspect, the
level of TNF-.alpha. in oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is about 25% or less of
the level of TNF-.alpha. present in conventionally stored blood
after 40 days.
[0080] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of epidermal growth factor (EGF) compared to
conventionally stored whole blood. In an aspect, the level of EGF
in oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) is about half the level of EGF
present in conventionally stored blood after 21 days, normalized to
the hemoglobin concentration. In an aspect, the level of EGF in
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is about 25% or less of the level of EGF
present in conventionally stored blood after 40 days.
[0081] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of soluble CD40 ligand (sCD40L) compared to
conventionally stored whole blood. In an aspect, the level of
sCD40L in oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) is about half the level of
sCD40L present in conventionally stored blood after 21 days,
normalized to the hemoglobin concentration. In an aspect, the level
of sCD40L in oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is about 25% or less of
the level of sCD40L present in conventionally stored blood after 40
days.
[0082] In certain aspects, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
reduced levels of platelet-derived growth factor (PDGF) compared to
conventionally stored whole blood. In an aspect, the level of PDGF
in oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) is about half the level of PDGF
present in conventionally stored blood after 21 days, normalized to
the hemoglobin concentration. In an aspect, the level of PDGF in
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is about 25% or less of the level of PDGF
present in conventionally stored blood after 40 days.
[0083] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that provides a reduced inflammatory
response when transfused into a patient compared to conventionally
stored oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT).
[0084] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB) that provides blood
products having higher RBC deformability compared to conventionally
stored blood products. In certain aspects, the blood product is a
whole blood product. In another aspect, the blood product is
leukoreduced whole blood. In another aspect, the blood product is
leukoreduced and platelet reduced whole blood. In a further aspect,
the blood product is leukoreduced packed red blood cells or leuko-
and platelet-reduced packed red blood cells.
[0085] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB) that has coagulation
parameters that are at least 75% of the coagulation parameter of
conventionally stored whole blood as measured by
thromboelastography (TEG). In an aspect, the TEG coagulation
parameter of the oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is equivalent to
conventionally stored blood. In yet another aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a TEG coagulation parameter that is greater than
the TEG coagulation parameter of conventionally stored blood. In an
aspect, the TEG Angle is more than 40.degree.. In another aspect,
the TEG kinetics (K) is less than 5 mins. In another aspect the TEG
K is between 1 to 5 mins. In another aspect, the TEG maximum
amplitude (TEG MA) is more than 50 mm. In another aspect, the TEG
maximum amplitude (TEG MA) is less than 70 mm. In another aspect,
the TEG maximum amplitude (TEG MA) is between 30 to 65 mm. In
another aspect, the TEG reaction time (TEG R) is less than 10 mins.
In another aspect, the TEG reaction time (TEG R) is less than 8
mins. In another aspect, the TEG reaction time (TEG R) is at least
3 mins. In a further aspect, the TEG reaction time (TEG R) is
between 4 to 8 mins.
[0086] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB) that has coagulation
parameters that are at least 75% of the coagulation parameter of
conventionally stored whole blood as measured by prothrombin time
(PT). In an aspect, the PT of the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) is
equivalent to conventionally stored blood. In yet another aspect,
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) has a PT that is greater than the PT of
conventionally stored blood. In a further aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB) has a PT of less than 15
seconds. In another aspect, oxygen reduced leukoreduced whole blood
(OR-LRWB) has a PT of more than 5 seconds. In another aspect,
oxygen reduced leukoreduced whole blood (OR-LRWB) or has a PT of 10
to 15 seconds.
[0087] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has coagulation parameters that are
at least 75% of the coagulation parameter of conventionally stored
whole blood as measured by partial thromboplastin time (PTT). In an
aspect, the PTT of the oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is equivalent to
conventionally stored blood. In yet another aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a PTT that is greater than the PTT of
conventionally stored blood. In a further aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a PTT that is greater than 25 seconds. In
another aspect, oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has a PTT that is less
than 40 seconds. In another aspect, oxygen reduced leukoreduced
whole blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
a PTT that is between 32 to 42 seconds.
[0088] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that have whole blood (OR-WB) that has
coagulation parameters that are at least 75% of the coagulation
parameter of conventionally stored whole blood as measured by the
level of fibrinogen activity. In an aspect, the fibrinogen activity
of the oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) is equivalent to conventionally
stored blood. In yet another aspect, oxygen reduced leukoreduced
whole blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
a fibrinogen activity that is greater than the fibrinogen activity
of conventionally stored blood. In a further aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a fibrinogen level that is at least 200 mg/ml.
In another aspect, oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has a fibrinogen level
that is at most 400 mg/ml. In another aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a fibrinogen level that is from 250 to 350
mg/ml. In another aspect, oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has a fibrinogen level
that is from 250 to 300 mg/ml.
[0089] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that have whole blood (OR-WB) that has
coagulation parameters that are at least 75% of the coagulation
parameter of conventionally stored whole blood as measured by
D-dimer analysis. In an aspect, the D-dimer value of the oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) is equivalent to conventionally stored
blood. In yet another aspect, oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
a D-dimer value that is greater than the D-dimer value of
conventionally stored blood.
[0090] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that have whole blood (OR-WB) that has
coagulation parameters that are at least 75% of the coagulation
parameter of conventionally stored whole blood as measured by a
thrombin generation assay. In an aspect, the thrombin generation
value of the oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is equivalent to
conventionally stored blood. In yet another aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a thrombin generation value that is greater than
the thrombin generation value of conventionally stored blood.
[0091] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has platelet function parameters that
are at least 75% of the platelet function parameters of
conventionally stored whole blood as measured by a platelet
aggregometer. In an aspect, the platelet function parameters of the
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) are equivalent to conventionally stored
blood. In yet another aspect, oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
platelet function parameters that are greater than the platelet
function parameters of conventionally stored blood.
[0092] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has levels of clotting factor that
are at least 75% of the levels of clotting factor in conventionally
stored blood. In an aspect, the level of clotting factors is
equivalent to that of conventionally stored blood. In yet other
aspects, oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) has a platelet function
parameter that is greater than the platelet function parameter of
conventionally stored blood. Not to be limited by theory, it is
thought that oxidative degradation of clotting factors is prevented
or reduced in oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) and provides for higher
levels of clotting factor activity. Methods to evaluate the effect
of treatments on coagulability are known in the art, for example as
described by Pidcoke et al., "Primary hemostatic capacity of whole
blood: a comprehensive analysis of pathogen reduction and
refrigeration effects over time," Transfusion 53:137S-149S
(2013).
[0093] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has a level of Factor V that has a
specific activity that is at least 75% of the level of Factor V
activity present in conventionally stored blood. In an aspect, the
specific activity of Factor V is equivalent to that of
conventionally stored blood. In yet other aspects, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a specific activity of Factor V that is greater
than the platelet function parameter of conventionally stored
blood. Methods of measuring the specific activity of Factor V are
known in the art.
[0094] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has a level of Factor V that has a
specific activity that is at least 75% of the level of Factor VIII
activity present in conventionally stored blood. In an aspect, the
specific activity of Factor VIII is equivalent to that of
conventionally stored blood. In yet other aspects, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a specific activity of Factor VIII that is
greater than the platelet function parameter of conventionally
stored blood. Methods of measuring the specific activity of Factor
VIII are known in the art. In one aspect, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a specific activity of Factor V that is less
than 40% after 21 days of storage.
[0095] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has a level of antithrombin (AT) that
has a specific activity that is at least 75% of the level of AT
activity present in conventionally stored blood. In an aspect, the
specific activity of AT is equivalent to that of conventionally
stored blood. In yet other aspects, oxygen reduced leukoreduced
whole blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) has
a specific activity of AT that is greater than the platelet
function parameter of conventionally stored blood. Methods of
measuring the specific activity of AT are known in the art.
[0096] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has a level of Factor XIV
(autoprothrombin IIA or Protein C) that has a specific activity
that is at least 75% of the level of Factor XIV activity present in
conventionally stored blood. In an aspect, the specific activity of
Factor XIV is equivalent to that of conventionally stored blood. In
yet other aspects, oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has a specific activity
of Factor XIV that is greater than the platelet function parameter
of conventionally stored blood. Methods of measuring the specific
activity of Factor XIV are known in the art.
[0097] The present disclosure provides for, and includes, oxygen
reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) that has a level of Von Willebrand factor
(vWF) that has a specific activity that is at least 75% of the
level of vWF activity present in conventionally stored blood. In an
aspect, the specific activity of vWF is equivalent to that of
conventionally stored blood. In yet other aspects, oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) has a specific activity of vWF that is greater than
the platelet function parameter of conventionally stored blood. In
another aspect, oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has a specific activity
of vWF that is less than the platelet function parameter of
conventionally stored blood. Methods of measuring the specific
activity of vWF are known in the art.
[0098] The present disclosure provides for, and includes, methods
to extend the stored shelf life of whole blood from the current 2
weeks to 3 weeks and beyond. The oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) of
the present disclosure provides for patient outcomes at three weeks
that are equivalent to patient outcome provided by whole blood that
has been stored for two weeks under conventional conditions.
[0099] As provided herein, oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has decreased side
effects for transfusion recipients as compared to conventionally
stored blood. In an aspect, oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has a decreased
inflammatory response after two weeks storage as compared to
conventionally stored blood. In other aspects, the inflammatory
response is reduced relative to conventionally stored blood after
three weeks. In an aspect, the oxygen reduced leukoreduced whole
blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT) can
be stored for more than three weeks and retain the levels of
inflammatory response as compared to conventionally stored blood
after two weeks.
[0100] As provided herein, oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has decreased side
effects for transfusion recipients as compared to conventionally
stored blood. In an aspect, oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) has a decreased immune
modulation after two weeks storage as compared to conventionally
stored blood. In other aspects, the immune modulation is reduced
relative to conventionally stored blood after three weeks. In an
aspect, the oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) can be stored for more
than three weeks and retain the levels of immune modulation as
compared to conventionally stored blood after two weeks.
[0101] The methods and whole blood products of the present
disclosure provide improved patient outcomes when transfused. In
particular, oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) provides for improved
survival in cancer patients when provided in perioperative
transfusions. In a certain aspect, the oxygen reduced leukoreduced
whole blood (OR-LRWB), oxygen reduced leukoreduced whole blood with
platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB), or oxygen and carbon dioxide
reduced leukoreduced whole blood with platelets (OCR-LRWB+PLT)
provides for reduced mortality and improved survival when provided
perioperatively to pancreatic cancer patients. Not to be limited by
theory, the reduced mortality is the result of the combination of
reduced levels of cytokines and improved oxygen transport and
delivery that results from increased levels of 2,3-DPG and ATP.
[0102] In an aspect, the blood for transfusion to a cancer patient
in need thereof has a reduced level of the cytokine regulated on
activation, normal T cell expressed and secreted (RANTES). In an
aspect, the RANTES level is equivalent to the level of RANTES
present at the beginning of storage. In another aspect, the RANTES
level of oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT),
oxygen and carbon dioxide reduced leukoreduced whole blood
(OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT) is less than the level of
RANTES present in conventionally stored blood. In an aspect, the
level of RANTES is less than the level of RANTES present in
conventionally stored blood throughout the storage period. In other
aspects, RANTES does not increase during storage.
[0103] In an aspect, the blood for transfusion to a cancer patient
in need thereof has a reduced level of a CC chemokine that is an
eosinophil chemotactic protein, eotaxin. In an aspect, the eotaxin
having a reduced level in oxygen reduced leukoreduced whole blood
(OR-LRWB), oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT) is eotaxin-1, also known
as C--C motif chemokine 11. In an aspect the eotaxin level is
equivalent to the level of eotaxin present at the beginning of
storage. In another aspect, the eotaxin level of oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) is less than the level of eotaxin present in
conventionally stored blood. In an aspect, the level of eotaxin is
less than the level of eotaxin present in conventionally stored
blood throughout the storage period. In other aspects, eotaxin does
not increase during storage.
[0104] The methods and whole blood products of the present
disclosure provide for reduced multiple organ dysfunction syndrome
and improved patient outcomes when transfused. In particular,
oxygen reduced leukoreduced whole blood (OR-LRWB), oxygen reduced
leukoreduced whole blood with platelets (OR-LRWB+PLT), oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB), or
oxygen and carbon dioxide reduced leukoreduced whole blood with
platelets (OCR-LRWB+PLT) provides for reduced multiple organ
dysfunction syndrome in trauma patients when provided in
perioperative transfusions. In a certain aspect, the oxygen reduced
leukoreduced whole blood (OR-LRWB), oxygen reduced leukoreduced
whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide
reduced leukoreduced whole blood (OCR-LRWB), or oxygen and carbon
dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT) provides for reduced multiple organ dysfunction
syndrome in trauma patients when provided during emergency
treatment.
[0105] The present disclosure provides for, and includes, a method
of preparing oxygen reduced leukoreduced whole blood comprising
obtaining a unit of whole blood comprising an anticoagulant,
filtering the whole blood to produce leukoreduced whole blood,
depleting the leukoreduced whole blood of oxygen, and storing the
oxygen reduced leukoreduced whole blood under anaerobic
conditions.
[0106] The present disclosure provides for, and includes, a method
of preparing oxygen reduced leukoreduced whole blood that has a
pre-storage oxygen saturation of (SO.sub.2) of 30% or less. Whole
blood obtained from a donor using venipuncture has an oxygen
saturation ranging from about 30% to about 70% saturated oxygen
(SO.sub.2). In certain aspects, the SO.sub.2 is reduced to 25% or
less. In certain aspects, the SO.sub.2 is reduced to 20% or less.
In certain aspects, the SO.sub.2 is reduced to 15% or less. In
other aspects, the SO.sub.2 is reduced to 10% or less. In yet other
aspects, the SO.sub.2 is reduced to 5% or less.
[0107] Also provided for and included in the present disclosure are
compositions and methods to prepare compositions of oxygen reduced
and carbon dioxide reduced leukoreduced whole blood. In certain
aspects, the SO.sub.2 value is 20% or less and the partial pressure
of carbon dioxide is less than 60 mmHg. In other aspects, the
partial pressure of carbon dioxide is between 10 and 60 mmHg. In
another aspect, the partial pressure of carbon dioxide is between
20 and 40 mmHg. Also included are whole blood compositions and
methods that provide for an SO.sub.2 of 15% or less and a partial
pressure of carbon dioxide of between 10 and 60 mmHg. In another
aspect, the methods and compositions include whole blood products
having an SO.sub.2 of 15% or less and a partial pressure of carbon
dioxide of between 20 and 40 mmHg. In yet another aspect, the blood
compositions and methods of the present disclosure have an SO.sub.2
of 10% or less and a partial pressure of carbon dioxide of between
10 and 60 mmHg. In other aspects, the blood compositions and
methods of the present disclosure have an SO.sub.2 of 10% or less
and a partial pressure of carbon dioxide of between 20 and 40 mmHg.
In yet further aspects, the blood compositions and methods of the
present disclosure have an SO.sub.2 of 5% or less and a partial
pressure of carbon dioxide of between 10 and 60 mmHg. In other
aspects, the blood compositions and methods of the present
disclosure have an SO.sub.2 of 5% or less and a partial pressure of
carbon dioxide of between 20 and 40 mmHg.
[0108] Also provided for and included in the present disclosure,
are compositions and methods to prepare compositions of oxygen
reduced and carbon dioxide reduced leukoreduced whole blood. In
certain aspects, the SO.sub.2 value is 20% or less and the partial
pressure of carbon dioxide is between 1 and 60 mmHg. In other
aspects, the partial pressure of carbon dioxide is between 10 and
60 mmHg. In another aspect, the partial pressure of carbon dioxide
is between 20 and 40 mmHg or 1 and 20 mmHg. Also included are whole
blood compositions and methods that provide for an SO.sub.2 of 15%
or less and a partial pressure of carbon dioxide of between 10 and
60 mmHg. In certain aspects, the SO.sub.2 value is 15% or less and
the partial pressure of carbon dioxide is between 1 and 60 mmHg. In
another aspect, the methods and compositions include whole blood
products having an SO.sub.2 of 15% or less and a partial pressure
of carbon dioxide of between 20 and 40 mmHg or 1 and 20 mmHg. In
yet another aspect, the blood compositions and methods of the
present disclosure have an SO.sub.2 of 10% or less and a partial
pressure of carbon dioxide of between 1 and 60 mmHg or 10 and 60
mmHg. In other aspects, the blood compositions and methods of the
present disclosure have an SO.sub.2 of 10% or less and a partial
pressure of carbon dioxide of between 20 and 40 mmHg or 1 and 20
mmHg. In yet further aspects, the blood compositions and methods of
the present disclosure have an SO.sub.2 of 5% or less and a partial
pressure of carbon dioxide of between 1 and 60 mmHg or 10 and 60
mmHg. In other aspects, the blood compositions and methods of the
present disclosure have an SO.sub.2 of 5% or less and a partial
pressure of carbon dioxide of between 20 and 40 mmHg or 1 and 20
mmHg.
[0109] Notably, and as evidenced in FIGS. 2A, 2B, 3A and 3B, the
ATP level in stored oxygen reduced blood depends on the partial
pressure of CO.sub.2. Specifically, depletion of oxygen to about
10% SO.sub.2 and carbon dioxide to about 25 mmHg results in
increased 2,3-DPG levels that persist beyond 21 days, while ATP
decreases to a level of about one half of the initial value. See
FIGS. 2G and 3D. Accordingly, the present disclosure provides for,
and includes the depletion of oxygen to SO.sub.2 levels of about 5%
and the depletion of carbon dioxide to a partial pressure of about
30 to 40 mmHg to yield oxygen and carbon dioxide reduced whole
blood that has increased levels of 2,3-DPG and that retains at
least 50% of the initial concentration of ATP through day 20. In
other aspects, the partial pressure of CO.sub.2 can be adjusted to
retain ATP levels that are at least 75% of the initial ATP value.
Adjustment of the level of CO.sub.2 can be experimentally
determined by one of ordinary skill in the art in view of the
present disclosure.
[0110] Prolonged hypothermic storage under conventional conditions
is known to impair deformability of stored RBCs, potentially
compromising their ability to perfuse microvascular networks and
deliver oxygen to tissues and vital organs upon transfusion. It is
thought that oxidative damage may be a primary contributor to the
loss of RBC biomechanical function; therefore, storing RBCs under
oxygen reduced (OR) and oxygen and carbon dioxide reduced (OCR)
conditions ameliorates oxidative damage, thereby preserving native
rheological properties better than conventional (aerobic) storage.
For this study, we utilized an in vitro microfluidic system that
recapitulates an in vivo microvascular capillary bed to demonstrate
the effects of reduced oxygen on stored cells.
[0111] The present disclosure provides for, and includes, methods
for managing a blood bank that improves the availability of blood
products for trauma victims and patients that require multiple
transfusions and provides conserving the overall blood resources.
The component blood products can be prepared from stored whole
blood of the present application and used for transfusions or
incorporated into massive transfusion kits. In addition to the
improved blood chemistries (low hemolysis, improved 2,3-DPG etc.),
the methods provide for improved hemostasis and improved
deformability.
[0112] In aspects according the present specification, the method
provides for maintaining an inventory of blood units comprising
oxygen reduced whole blood and an anticoagulant as described above,
providing one or more of the blood units from the inventory for
treatment of a patient and recycling the blood units from the
inventory to prepare component separated oxygen reduced blood units
including oxygen reduced plasma and oxygen reduced leukoreduced
packed red blood cells with platelets (OR-LRpRBC+PLT). In aspects,
the anticoagulant comprises citrate-phosphate-dextrose (CPD),
citrate-phosphate-dextrose with adenine (CPDA-1), or CP2D.
[0113] In an aspect, the specification provides for a method for
maintaining an inventory of blood units comprising oxygen and
carbon dioxide reduced leukoreduced whole blood and an
anticoagulant as described above, providing one or more of the
blood units from the inventory for treatment of a patient and
recycling the blood units from the inventory to prepare component
separated oxygen reduced blood units including oxygen and carbon
dioxide reduced plasma and oxygen and carbon dioxide reduced
leukoreduced packed red blood cells with platelets
(OCR-LRpRBC+PLT). In aspects, the anticoagulant comprises
citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose with
adenine (CPDA-1), or Anticoagulant Citrate Phosphate Double
Dextrose (CP2D).
[0114] The specification further provides for preparing one or more
massive transfusion kits as described below that include the oxygen
reduced plasma, oxygen reduced leukoreduced packed red blood cells
with platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide reduced
plasma and oxygen and carbon dioxide reduced leukoreduced packed
red blood cells with platelets (OCR-LRpRBC+PLT).
[0115] In aspects according to the invention, the unused blood in
the inventory is recycled after a time period. In certain aspects
where the anticoagulant is CPD, the blood units are recycled prior
to three weeks of storage. In other aspects where the anticoagulant
is CPDA-1, the blood units are recycled prior to five weeks of
storage. In yet other aspects, the blood units are recycled after 2
weeks, or less. In an aspect, blood unit recycling occurs between 2
days and 1 week. In another aspect, recycling occurs between 2 days
and two weeks. In some aspects, recycling occurs between 1 week and
2 weeks. The timing of recycling can be varied consistent with the
turnover and needs of the blood facility.
[0116] While the recycling process is preferably performed under
anaerobic conditions, the process may also be performed under
aerobic conditions. Aerobic conditions may provide a cost savings,
but may also be indicated in facilities with higher turnover. In
high turnover facilities, the recovered blood components may be
used soon after the recycling process and further storage of the
blood under anaerobic conditions may provide little additional
benefit.
[0117] The method for managing a blood bank further provide for the
preparation of a massive transfusion kit as described in detail
below.
[0118] The present disclosure provides for, and includes, methods
to provide a supply of blood products for transfusion medicine
comprising depleting oxygen from leukoreduced whole blood to
prepare oxygen reduced leukoreduced whole blood (OR-LRWB+PLT),
storing the oxygen reduced leukoreduced whole blood (OR-LRWB+PLT)
for a time period and providing said stored blood to a patient in
need thereof. In certain aspects, the leukoreduction step includes
platelet reduction to produce oxygen reduced leukoreduced whole
blood (OR-LRWB).
[0119] The present disclosure provides for, and includes, methods
to provide a supply of blood products for transfusion medicine
comprising depleting oxygen and carbon dioxide from leukoreduced
whole blood to prepare oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB+PLT), storing the oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB+PLT) for
a time period and providing said stored blood to a patient in need
thereof. In certain aspects, the leukoreduction step includes
platelet reduction to produce oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB).
[0120] The present disclosure provides for, and includes, methods
to provide a supply of blood products for transfusion medicine
comprising depleting oxygen from leukoreduced whole blood to
prepare oxygen reduced leukoreduced whole blood (OR-LRWB+PLT),
storing the oxygen reduced leukoreduced whole blood (OR-LRWB+PLT)
for a time period and preparing oxygen reduced leukoreduced packed
red blood cells with platelets (OR-LRpRBC+PLT). In certain aspects,
the leukoreduction step includes platelet reduction to produce
oxygen reduced leukoreduced packed red blood cells (OR-LRpRBC).
[0121] The present disclosure provides for, and includes, methods
to provide a supply of blood products for transfusion medicine
comprising depleting oxygen and carbon dioxide from leukoreduced
whole blood to prepare oxygen and carbon dioxide reduced
leukoreduced whole blood (OCR-LRWB+PLT), storing the oxygen and
carbon dioxide reduced leukoreduced whole blood (OCR-LRWB+PLT) for
a time period and preparing oxygen and carbon dioxide reduced
leukoreduced packed red blood cells with platelets
(OCR-LRpRBC+PLT). In certain aspects, the leukoreduction step
includes platelet reduction to produce oxygen and carbon dioxide
reduced leukoreduced packed red blood cells (OCR-LRpRBC).
[0122] As provided herein, the OR-LRpRBC+PLT, OR-LRpRBC,
OCR-LRpRBC+PLT, and OCR-LRpRBC may be returned to the store of
blood products for supply and stored for a time period until
required for use by a patient. In aspects of the present
disclosure, the total time for storage, either as a whole blood
product, or as a packed RBC product may be up to six weeks. In some
aspects, the second storage period is between 2 to 4 weeks.
[0123] The methods for providing a supply of blood products
include, and provide, for depleting either oxygen or oxygen and
carbon dioxide. The oxygen levels for the methods to provide a
supply of blood products are discussed in detail above. In certain
aspects, the SO.sub.2 value is reduced to 20% or less and the
partial pressure of carbon dioxide is less than 60 mmHg. In other
aspects, the partial pressure of carbon dioxide is between 10 and
60 mmHg. In another aspect, the partial pressure of carbon dioxide
is between 20 and 40 mmHg. Also included are methods that provide
for an SO.sub.2 of 15% or less and a partial pressure of carbon
dioxide of between 10 and 60 mmHg. In another aspect, the methods
provide blood products having an SO.sub.2 of 15% or less and a
partial pressure of carbon dioxide of between 20 and 40 mmHg. In
yet another aspect, the methods of the present disclosure provide
blood products having an SO.sub.2 of 10% or less and a partial
pressure of carbon dioxide of between 10 and 60 mmHg. In other
aspects, the methods for providing a supply of blood products
provide for an SO.sub.2 of 10% or less and a partial pressure of
carbon dioxide of between 20 and 40 mmHg. In yet further aspects,
the methods provide an SO.sub.2 of 5% or less and a partial
pressure of carbon dioxide of between 10 and 60 mmHg. In other
aspects, the methods provide an SO.sub.2 of 5% or less and a
partial pressure of carbon dioxide of between 20 and 40 mmHg.
[0124] The present disclosure provides for, and includes, a new
blood composition obtained during the blood component recovery
process of OR-LRWB+PLT and OCR-LRWB+PLT. As provided above, while
conventional whole blood products have an FDA approved shelf life
(3 weeks for WB in CPD and 5 weeks in CPDA1), clinicians who use WB
limit its shelf life from between 2 and 14 days. In conventional
storage, the blood is often discarded. In the present disclosure,
the OR-LRWB+PLT and OCR-LRWB+PLT can be processed using
conventional component separation methods modified for retaining
the blood in the OR or OCR depleted state. In general, methods are
modified to incorporate oxygen and oxygen and carbon dioxide
impermeable barriers to the components and incorporating features
to prevent oxygen ingress. Suitable approaches can be found, for
example in International Patent Application No. PCT/US2016/021794,
filed Mar. 10, 2016, and International Patent Application No.
PCT/US2016/029069, filed Apr. 22, 2016, both of which are hereby
incorporated by reference in their entireties.
[0125] In aspects according the present disclosure, a blood
composition is provided that comprises oxygen reduced packed red
blood cells and platelets having less than 1.times.10.sup.5/L white
blood cells. Such compositions are obtainable from OR-LRWB+PLT and
OCR-LRWB+PLT. In an aspect, the level of white blood cells is less
than 1.times.10.sup.4/L white blood cells. In aspects according to
the present disclosure, the oxygen saturation of the oxygen reduced
leukoreduced packed red blood cells with platelets (OR-LRpRBC+PLT)
is less than 30%. In an aspect, the oxygen saturation of the oxygen
reduced leukoreduced packed red blood cells with platelets
(OR-LRpRBC+PLT) is less than 20%. In an aspect, the oxygen
saturation of the oxygen reduced leukoreduced packed red blood
cells with platelets (OR-LRpRBC+PLT) is less than 10%. In a further
aspect, the oxygen saturation of the oxygen reduced leukoreduced
packed red blood cells with platelets (OR-LRpRBC+PLT) is less than
5%.
[0126] The present disclosure provides for, and includes, oxygen
and carbon dioxide reduced leukoreduced packed red blood cells with
platelets (OCR-LRpRBC+PLT) having less than 30% SO.sub.2 and a
storage partial pressure of carbon dioxide of less than 60 mmHg. In
an aspect, the OCR-LRpRBC+PLTs have an oxygen saturation of less
than 30% and a storage partial pressure of carbon dioxide between
20 and 40 mmHg. In an aspect, the OCR-LRpRBC+PLTs have an oxygen
saturation of less than 30% and a storage partial pressure of
carbon dioxide between 0 and 20 mmHg. In an aspect, the
OCR-LRpRBC+PLTs have an oxygen saturation of less than 20% and a
storage partial pressure of carbon dioxide of less than 60 mmHg. In
an aspect, the OCR-LRpRBC+PLTs have an oxygen saturation of less
than 20% and a storage partial pressure of carbon dioxide between
20 and 40 mmHg. In an aspect, the OCR-LRpRBC+PLTs have an oxygen
saturation of less than 20% and a storage partial pressure of
carbon dioxide between 0 and 20 mmHg. In another aspect, the
OCR-LRpRBC+PLTs have an oxygen saturation of less than 15% and a
storage partial pressure of carbon dioxide of less than 60 mmHg. In
an aspect, the OCR-LRpRBC+PLTs have an oxygen saturation of less
than 15% and a storage partial pressure of carbon dioxide between
20 and 40 mmHg. In an aspect, the OCR-LRpRBC+PLTs have an oxygen
saturation of less than 15% and a storage partial pressure of
carbon dioxide between 0 and 20 mmHg. In another aspect, the
OCR-LRpRBC+PLTs have an oxygen saturation of less than 10% and a
storage partial pressure of carbon dioxide of less than 60 mmHg. In
an aspect, the OCR-LRpRBC+PLTs have an oxygen saturation of less
than 10% and a storage partial pressure of carbon dioxide between
20 and 40 mmHg. In an aspect, the OCR-LRpRBC+PLTs have an oxygen
saturation of less than 10% and a storage partial pressure of
carbon dioxide between 0 and 20 mmHg. In other aspects, the
OCR-LRpRBC+PLTs has an oxygen saturation of less than 5% and a
storage partial pressure of carbon dioxide of less than 60 mmHg. In
an aspect, the OCR-LRpRBC+PLTs have an oxygen saturation of less
than 5% and a storage partial pressure of carbon dioxide between 20
and 40 mmHg. In an aspect, the OCR-LRpRBC+PLTs have an oxygen
saturation of less than 5% and a storage partial pressure of carbon
dioxide between 0 and 20 mmHg.
[0127] The oxygen reduced leukoreduced packed red blood cells with
platelets (OR-LRpRBC+PLT) and oxygen and carbon dioxide reduced
leukoreduced whole blood with platelets (OCR-LRWB+PLT) usually
further comprise an additive solution. Suitable additive solutions
according to the present disclosure include AS-1, AS-3
(Nutricel.RTM.), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5,
EAS61, OFAS1, OFAS3, and combinations thereof. In an aspect, the
additive solution is added at the time of component separation. In
an aspect, the additive solution is AS-1. In another aspect, the
additive solution is AS-3. In other aspects, the additive solution
is SAGM.
[0128] The methods and compositions of the present disclosure
provide for and include the preparation of `massive transfusion
kits` (MTKs) having improved properties to kits prepared from
conventional components. The massive transfusion kits of the
present disclosure can be prepared in various configurations
depending on the clinical needs. The MTKs of the present disclosure
are stored under oxygen free or oxygen and carbon dioxide free
conditions until being prepared for use. The OR and OCR conditions
can be maintained by sealing in an impermeable enclosure either
with, or without, an appropriate sorbent material. The MTKs of the
present disclosure may be re-oxygenated prior to use, or used
directly. In general, the specification provides for massive
transfusion kits optimized to deliver RBCs having improved 2,3-DPG
levels. Such kits are prepared from component blood products
obtained from oxygen and carbon dioxide reduced leukoreduced whole
blood with platelets (OCR-LRWB+PLT). Alternatively, a kit may be
prepared using component blood products obtained from oxygen
reduced leukoreduced whole blood with platelets (OR-LRWB+PLT) to
produce kits having higher levels of ATP. Kits prepared using the
methods of the present specification provide platelets suitable for
hemostasis together with the oxygen reduced stored red blood cells.
Thus, massive transfusion kits of the present specification can
increase the availability of platelets without additional dilution
while further providing RBCs of higher quality (e.g., more
deformable, more 2,3-DPG, fewer storage lesions). Importantly, the
recovery of blood components from the oxygen reduced whole blood of
the present disclosure increases the availability of transfusion
products for trauma victims and saves and conserves a valuable and
limited resource. As discussed above, conventional massive
transfusion kits include a volume of plasma, a volume of pRBCs, and
volume of platelets in a 1:1:1 ratio wherein the amounts of the
three components correspond to a unit `reconstituted blood` when
transfused serially or in parallel to a patient in need.
Reconstituted blood does not directly correspond to whole blood
which does not include an additive solution and has higher levels
of anti-coagulant. Further reconstituted blood typically includes a
larger volume than a typical unit of whole blood. The reconstituted
blood of the present disclosure is improved over the conventional
reconstituted blood as it provides additional platelets in the pRBC
fraction (e.g., either oxygen and carbon dioxide reduced
leukoreduced packed red blood cells with platelets (OCR-LRpRBC+PLT)
and oxygen reduced leukoreduced packed red blood cells with
platelets (OR-LRpRBC+PLT)). Such cold stored platelets undergo
modification commonly known as platelet storage lesions (PSLs) and
cold stored platelets are quickly removed from circulation in the
body. Importantly, cold stored platelets retain the ability to
aggregate and have been reported to have increased aggregations and
resistance to disaggregation. Accordingly, blood components
obtained from the oxygen reduced whole blood of the present
specification provide additional benefits during trauma
transfusion, either alone or in combination with conventional
platelets.
[0129] The present disclosure provides for, and includes, a massive
transfusion kit comprising a volume of oxygen reduced leukoreduced
packed red blood cells with platelets (OR-LRpRBC+PLT) or oxygen and
carbon dioxide reduced leukoreduced whole blood with platelets
(OCR-LRWB+PLT), or combinations thereof. In an aspect, a massive
transfusion kit provides a volume of plasma and a volume of
LRpRBC+PLT. In an aspect, the volume of plasma and a volume of
LRpRBC+PLT is 1:1. In other aspects, the ratio of plasma to
LRpRBC+PLT is between 1:1 and 1:2 by volume. In an aspect, the
ratio of plasma to LRpRBC+PLT is about 1:2 by volume.
[0130] The present disclosure provides for, and includes, massive
transfusion kits that include additional platelets together with
the plasma and oxygen reduced leukoreduced packed red blood cells
with platelets (OR-LRpRBC+PLT) or oxygen and carbon dioxide reduced
leukoreduced whole blood with platelets (OCR-LRWB+PLT).
[0131] The massive transfusion kits of the present disclosure
provide for a volume of plasma. The plasma of the MTKs can be
either fresh plasma or thawed fresh frozen plasma (FFP). The
specification provides for obtaining the plasma for MTKs from
either conventional sources (e.g., non-oxygen reduced) or from
oxygen reduced or oxygen and carbon dioxide reduces sources. In an
aspect, the plasma for an MTK of the present disclosure may be
obtained from oxygen reduced leukoreduced whole blood (OR-LRWB),
oxygen reduced leukoreduced whole blood with platelets
(OR-LRWB+PLT), oxygen and carbon dioxide reduced leukoreduced whole
blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukoreduced
whole blood with platelets (OCR-LRWB+PLT). Not to be limited by
theory, the plasma obtained from the oxygen reduced sources will
have lower levels of storage lesions, including for example, lower
levels of cytokines, isoprostane, and microparticles. As provided
herein, MTKs having plasma, platelets and pRBCs according to the
present specification are provided at a ratio of between 1:1:1 or
1:1:2 by volume. It will be understood to one of ordinary skill in
the art that the MTKs of the present disclosure, like conventional
MTKs, are designed to provide an equivalent of a unit of blood. It
will be recognized that any arbitrary total volume may be selected
while maintaining the recited ratios necessary to be equivalent to
reconstituted blood.
EXAMPLES
Example 1: Cytokine, Cell-Free Hemoglobin, and Isoprostane
Accumulations in Packed Red Blood Cells During Anaerobic
Storage
[0132] Fifteen pRBC units are collected from normal healthy donors.
Each unit is split and stored as follows: one in standard blood
bank conditions (control), the other anaerobically (test) according
to methods described in Yoshida el al., "Anaerobic Storage of Red
Blood Cells in a Novel Additive Solution Improves In vivo
Recovery," Transfusion 49:458-64 (2008). At weeks 0, 1, 2, 3, and
6, samples are removed using a sterile connecting device from the
PRBC units. Plasma samples are frozen for the following assays:
single batch testing for 22 cytokines using the Procarta
Immunoassay Magnetic Bead kit, 8-isoprostane F.sub.2.alpha. via
mass spectrometric assay, and cell free hemoglobin via HemoCue
plasma/photometer (HemoCue AB, Angelholm, Sweden).
[0133] As shown in FIG. 1A, eotaxin reaches a statistically
significant difference at week 2 (86.6 pg/ml-control (c), 64.9-test
(t), p-value--0.00213, with statistical significance of p<0.05;
day 42 (292-c, 112-t; p=0.000). As shown in FIG. 1B, RANTES is
different at all time points, starting at day 3 (374.6-c, 55.1-t),
p-=0.00000; a very large difference is observed on day 42
(3371.6-c, 88.4-t; p<0.002). As shown in FIG. 1C, differences in
cell-free hemoglobin are seen at week 2 (96.0 mg/dl-c, 41.7-t),
p-=0.00001; day 42 (170-c, 63-t, p=0.0002). As shown in FIG. 1D,
storage day 3 shows differences in isoprostane (45.5 pg/ml-c,
32.1-t), p=0.00689; day 42 (101.9-c, 64.7-t, p=0.0048).
Example 2: Collection, Leukoreduction and Gas Depletion of Whole
Blood
[0134] A unit of blood is collected from a donor patient into
anticoagulant solution comprising either CPDA1 or CPDA according to
standard protocols, including collection of heparin tubs. The
collected blood containing anticoagulant is leukoreduced according
to manufacturer's instructions less than or equal to four hours
after the initial blood draw. Baseline ABL90 blood gas and
metabolic parameters are determined from the donor heparin tube and
the whole blood product according to standard procedures. See BSL
Handbook Procedure BSL-P024: Procedure Manual and Radiometer ABL90
FLEX Gas Analyzer instructions.
[0135] An anaerobic control is prepared from each unit of
leukoreduced blood by transferring 120 ml of LRWB/CPDA-1 or
LRWB/CPD into a 150 mL transfer bag, labeled as appropriate and
placed at room temperature (15.degree.-30.degree. C.).
[0136] The remainder of the LRWB LRWB/CPDA-1 or LRWB/CPD is
processed for oxygen or oxygen and carbon dioxide depletion by
transferring to a blood processing bag connected to a Sorin D100
and processed for 5 minutes at a flow rated of 700 ml/minute
without gas to generate a BOF processing control. 120 g of the
resulting BOF processed blood is transferred to a 300 ml transfer
bag that has been stored under anaerobic conditions and labeled BOF
processing control. The remainder of the LRWB/CPDA-1 or LRWB/CPD is
processed on the Sorin D100 at a peak flow rate of 700 ml/minute
with a gas flow rate of 3 L/min of a gas composition comprising 5%
CO.sub.2/95% N.sub.2 until the blood reaches .about.5% SO.sub.2
measuring blood gas values on a Radiometer ABL90 FLEX Gas Analyzer
at 3 to 5 minute intervals. To reduce carbon dioxide levels, the
gas mixture is switched to 100% N.sub.2 for 1 to 4 minutes until
SO.sub.2 reaches 5.+-.1% and pCO.sub.2 reaches 30.+-.3 mmHg,
monitoring blood gas values every 15-30 seconds to monitor
deoxygenation rate. 120 g of the resulting oxygen and carbon
dioxide reduced LRWB/CPDA-1 or LRWB/CPD is transferred to a 300 ml
transfer bag previously stored under anaerobic conditions as
described above and labeled ("C"). Further processing of the
LRWB/CPDA-1 or LRWB/CPD is performed on the Sorin D100 at a flow
rate of 700 ml/min with 99% N.sub.2 and 1% O.sub.2 until the
LRWB/CPDA-1 or LRWB/CPD reaches an SO.sub.2 of 5.+-.1% and
pCO.sub.2 reaches 7.+-.3 mmHg. 120 g of the resulting oxygen and
carbon dioxide reduced LRWB/CPDA-1 or LRWB/CPD is transferred to a
300 ml transfer bag previously stored under anaerobic conditions as
described above and labeled ("D"). Additional samples are processed
as described above using a new Sorin D100. Immediately following
the preparation of each sample, ABL90 blood gas levels are
determined according to manufacturer's instructions to establish
baseline SO.sub.2 and pCO.sub.2 levels (e.g., T.sub.0). See BSL
Handbook Procedures. Samples for cytokine analysis are collected
and stored at -80.degree. C. for later analysis.
[0137] All samples are analyzed as provided below at Example 6.
Example 3: Storage of Anaerobic Test Products
[0138] Oxygen reduced and oxygen and carbon dioxide reduced blood
in transfer bags are wrapped in mesh, secured with elastic and
placed in anaerobic canisters with 4 sorbent sachets (Mitsubishi,
SS-300). Canisters are sealed and the canister purged of air using
an Alicat Gas Processing System. See BSL Handbook Procedure
BSL-P040: Procedure for Placing Blood Products in Anaerobic Storage
in Canisters. Anaerobic and aerobic blood is placed in a Blood Bank
refrigerator at 1 to 6.degree. C. Canister gauges are monitored
daily to ensure that they read 5.+-.1 psi. Canisters that fall
below 2 psi are adjusted to standard procedures. See BSL Handbook
Procedure BSL-P040: Procedure for Placing Blood Products in
Anaerobic Storage in Canisters.
Example 4: Sample Testing
[0139] Samples are tested at indicated time points: day zero
(T.sub.0) post processing, day 1, week 1, week 2, and week 3.
Samples may be tested fresh or frozen for later testing as
appropriate for a given test. The testing includes a complete blood
count (CBC), Thromboelastography (TEG).
[0140] Prepare platelet rich plasma (PRP) for platelet aggregation
immediately per manufacturer's instructions.
[0141] Perform coagulation screening and additional assays per
manufacturer's instructions.
[0142] Prepare samples for cytokines immediately per manufacturer's
instructions.
Example 5: ATP Sampling and Measurement
[0143] Samples are processed for ATP measurement by
deproteinization and precipitation. 1 ml of sample (e.g., LRWB/CPD
or LRWB/CPDA-1 or samples described above) is precipitated with 1.0
ml ice cold trichloroacetic acid (TCA) (12% w/v) and vortexed for
15 to 30 seconds and incubated on ice for 5 minutes. Tubes
containing the TCA/Sample mixture are centrifuged at 3600 g for 5
minutes at 4.degree. C. Samples are immediately processed to
minimize exposure to TCA. The clarified supernate is transferred to
a pre-cooled tube and snap frozen on a dry ice alcohol bath and
stored at -70.degree. C.
Example 6: Improved Deformability in Stored RBCs that have been
Stored Under Oxygen Reduced Conditions
[0144] Nine (9) individual units of whole blood are obtained from
healthy consenting volunteers via a standard 500 mL blood donation.
Donated whole blood is processed into leukoreduced red blood cell
(LR-RBC) units according to standard AABB/FDA guidelines; the
resulting units are then split into two halves. One half of the
units are O.sub.2 and CO.sub.2 reduced as described in Examples 2
and 3.
[0145] The resulting samples are placed in anaerobic, hypothermic
storage, while the second half is placed in conventional, aerobic
hypothermic storage. Paired RBC units are stored in a blood bank
refrigerator and evaluated weekly for the entire duration of 6-week
storage. Prior to testing, the hematocrit of all RBC samples was
adjusted to 40% using normal saline (0.9% NaCl; RBC-S). The
deformability of the RBC-S at the beginning and during the study is
determined as described in International Patent Publication No. WO
2013/177339, published Nov. 28, 2013. High speed image sequences
(.about.150FPS) of the blood samples traversing artificial
microvascular network (AMVN) chip are recorded. The occlusion time,
the amount of time flow through the network that is obstructed by
non-deformable cells and the frequency at which blood profusion
through the network is interrupted (occlusion frequency) are
determined.
[0146] Overall bulk perfusion rates through the AMVN system are
consistently higher for O.sub.2 and CO.sub.2 controlled blood
compared to aerobically stored units and the total occlusion time
is consistently lower for oxygen reduced RBCs (Table 1). These
results suggest that reduction of oxygen levels in LR-RBC units
mitigates the deterioration of the biomechanical properties of the
red blood cell during hypothermic storage.
[0147] The oxygen depletion and storage process significantly
reduces the rate at which the rheological properties of RBCs
deteriorate during hypothermic storage, and is capable of
preserving more physiologically relevant biomechanical properties
of the red cells during storage. The improved deformability of the
RBCs combined with the benefits of whole blood transfusion
indicates that the preserved RBC function will improve retention of
RBCs post transfusion, and increased capability of transfused RBCs
to profuse the microvasculature.
TABLE-US-00001 TABLE 1 Perfusion rates of Blood Cells After Oxygen
Reduced Storage Week 0 Week 1 Week 2 Week 3 Week 4 Week 5 Week 6
MMCN O.sub.2 Reduced 203.8 .+-. 7.4 182.3 .+-. 13.0 167.1 .+-. 18.2
155.2 .+-. 12.3 133.2 .+-. 17.0 131.7 .+-. 14.1 122.7 .+-. 10.7
perfusion Conventional 200.4 .+-. 8.5 173.8 .+-. 10.6 160.0 .+-.
18.1 142.9 .+-. 8.8 122.3 .+-. 12.2 117.0 .+-. 12.5 106.2 .+-. 14.2
rate (pL/s) MMCN O.sub.2 Reduced 18.1 .+-. 8.5 36.6 .+-. 7.8 35.4
.+-. 10.3 45.9 .+-. 5.1 59.1 .+-. 8.5 60.1 .+-. 7.5 63.4 .+-. 2.1
plugging Conventional 21.3 .+-. 9.6 41.0 .+-. 10.0 42.1 .+-. 13.8
56.5 .+-. 2.9 67.3 .+-. 8.8 68.1 .+-. 7.6 71.3 .+-. 6.1 event time
(%) AMVN O.sub.2 Reduced 222.8 .+-. 12.5 206.7 .+-. 6.3 200.6 .+-.
3.5 186.4 .+-. 6.7 187.0 .+-. 12.7 176.5 .+-. 13.5 171.4 .+-. 13.7
perfusion Conventional 220.0 .+-. 14.9 200.8 .+-. 7.4 194.3 .+-.
8.8 182.3 .+-. 3.5 181.8 .+-. 12.0 171.9 .+-. 11.2 162.0 .+-. 11.7
rate (pL/s)
Example 7: Deoxygenation of Platelets does not Impede Hemostatic
Performance
[0148] Eight (8) units of whole blood (WB) are obtained from
healthy consenting volunteers via a standard 500 mL blood donation.
Donated whole blood is collected in CPDA-1 anticoagulant as
described in Example 2 (Research Blood Components, Inc.) and
leukoreduced with a platelet sparing filter (Imuflex.RTM. WB-SP)
(LRWB: Terumo Medical Corporation). The resulting filtered units
are then split into two halves. One half of the units are placed in
conventional, aerobic hypothermic storage, while the second half is
further divided and placed in anaerobic, hypothermic storage. The
anaerobically stored units are oxygen reduced (OR-LRWB) or oxygen
and carbon dioxide reduced (OCR-LRWB). The anaerobically stored
units are processed with the Sorin D100 membrane oxygenator to
yield anaerobic units with about 5% SO.sub.2 and about 35
mmHgpCO.sub.2. The resulting anaerobic units are placed in standard
PVC bags and stored in anaerobic canisters comprising oxygen
sorbent and Nitrogen gas. Paired leukoreduced platelet units are
evaluated weekly for the entire duration of 21 day storage as
described below.
[0149] The units are evaluated for metabolic parameters including
percent hemolysis (Plasma Low, Angelholm Sweden), ATP (DiaSys,
Flacht, Germany), and 2,3-DPG (Sigma-Aldrich, St. Louis, Mo.)
according to manufacturer's instructions. As shown in FIG. 4A,
reduced levels of ATP are maintained in stored OCR-LRWB but
increase in stored OR-LRWB compared to conventionally stored LRWB
(solid line). As shown in FIG. 4B, increased levels of 2,3-DPG were
maintained in stored OCR-LRWB and OR-LRWB compared to
conventionally stored LRWB, for up to 21 days. Further, as shown in
FIG. 4C, hemoloysis is not significantly changed when comparing
stored OR-LRWB and stored OCR-LRWB to conventionally stored LRWB
(solid line).
[0150] The conventionally stored LRWB and OCR-LRWB are assessed for
plasma coagulation parameters by evaluating Prothrombin Time (PT),
activated Partial Prothrombin time (aPTT), and the levels of
Fibrinogen and D-dimer. As shown in FIG. 5, the aPPT and PT were
slightly, but not significantly prolonged in conventionally stored
LRWB (solid line). Further, no evidence of coagulation activation
was observed as evidence by similar fibrinogen and D-dimer
levels.
[0151] The conventionally stored LRWB and OCR-LRWB are further
assessed for plasma clotting factors by determining the activity
levels for factors V, VIII, protein C activity, protein S activity
and von Willebrand Factor (vWF). Protein C and protein S analysis
are performed using the ACL TOP.RTM. (Instrumentation Laboratory)
and the STA-R Evolution coagulation Analyzer.RTM. (Diagnostica
Stago, Inc.), respectively, according to the manufacturers
instructions. As shown in FIG. 6, the levels of Factor V, Factor
VIII, Protein C activity, protein S activity, and vWF were not
significantly changed in anaerobic, hypothermic stored OCR-LRWB
(broken line) compared to conventionally stored WB (solid
line).
[0152] The conventionally stored LRWB and OCR-LRWB are further
assessed for coagulation using thromboelastography (TEG) with the
Haemoscope Thromboelastograph.RTM. analyzer (Haemonetics) according
to the manufacturer's instructions. As shown in FIGS. 7A to 7D, no
significant difference was observed for propagation (TEG Angle),
amplification (TEG K), maximum amplitude (TEG MA), or reaction time
(TEG R) in OCR-LRWB (broken line) compared to conventionally stored
LRWB (solid line).
[0153] While the present disclosure has been described with
reference to particular embodiments, it will be understood by those
skilled in the art that various changes may be made and equivalents
may be substituted for elements thereof without departing from the
scope of the present disclosure. In addition, many modifications
may be made to adapt a particular situation or material to the
teachings of the present disclosure without departing from the
scope of the present disclosure.
[0154] Therefore, it is intended that the present disclosure not be
limited to the particular embodiments disclosed as the best mode
contemplated for carrying out the present disclosure, but that the
present disclosure will include all embodiments falling within the
scope and spirit of the appended claims.
* * * * *