SAGM Blood Research Articles

SAGM blood for neonatal large volume transfusion

SAGM blood for neonatal large volume transfusion

Bacterial antigen-induced release of white cell- and platelet-derived bioactive substances in vitro.

Poor prognosis after resection of primary colorectal cancer may be related to the combination of perioperative blood transfusion and subsequent development of infectious complications. Various white cell- and platelet-derived cancer-growth substances may be involved in this process. Therefore, we studied the in vitro release of substances from white cells and platelets stimulated by bacterial antigens and supernatants from stored red-cell components. Eight units of whole blood (WB) and 8 U of buffy-coat-depleted red-cell (SAGM) blood were donated by healthy blood donors. Subsequently, one-half of each unit was leucocyte-depleted by filtration, and all 32 half-units were stored under standard conditions for 35 d. Just after storage, and on d 7, 21, and 35 during storage, aliquots of the supernatants were removed from the units and frozen at -80 degrees C. WB from other healthy donors was stimulated for 2 h with sodium chloride (controls), with Escherichia coli (E. coli) lipopolysaccharide (LPS) alone, or with LPS plus supernatants from the WB units (diluted 1:10), or from the SAGM units (diluted 1:20) stored for 0, 7, 21, or 35 d, respectively. Similar assays were performed using Staphylococcus aureus-derived protein A as a stimulatory antigen. The concentration of eosinophil cationic protein (ECP), myeloperoxidase (MPO), histamine (HIS), and plasminogen-activator inhibitor-1 (PAI-1) were determined in supernatants from the stored blood and in assay supernatants by using enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) methods. The extracellular concentration of ECP, MPO, and HIS increased significantly in a storage-time-dependent manner in nonfiltered WB and SAGM blood, and the increase was abrogated by prestorage leukofiltration. Similarly, PA-1 increased significantly in nonfiltered WB, and the increase was abrogated by prestorage leukofiltration. The supernatant concentrations of the four substances were significantly increased in LPS-stimulated (0.5-4 fold) and in protein A-stimulated (0.5-13.5-fold) assays compared with controls. The addition of supernatants from stored nonfiltered WB or SAGM blood significantly increased the assay supernatant of ECP, MPO, HIS, and PAU-1 concentrations storage-time-dependently in LPS-stimulated assays. Prestorage leukofiltration abrogated the additional effect of supernatants from stored blood. Similar results were observed for ECP and HIS through the addition of supernatants from stored blood to protein A-stimulated assays. Protein A stimulation did not lead to increased PA-1 release in assays diluted by supernatants from stored blood. However, the MPO concentrations were significantly (p = 0.004), and independent of storage time and leukofiltration, increased in protein A-stimulated assays diluted by supernatants from stored blood compared with sodium chloride dilution. Extracellular ECP, MPO, HIS, and PA-1 accumulate during storage of nonfiltered red-cell components, but the accumulation can be prevented by prestorage leukofiltration. In addition, bacterial antigens appear to induce significant release of the substances from white cells and platelets. Addition of supernatants from stored, nonfiltered WB and SAGM blood may increase the substance levels in a storage-time-dependent manner, and prestorage leukofiltration may prevent further increase by supernatants, except for MPO.

A reply

We thank you for the opportunity to respond to Dr Horsey's comments on our case report and to clarify any ambiguity. The blood preparation given was indeed red cells in an SAGM preparation but Dr Horsey's calculations are somewhat misleading. Each SAGM unit contains around 220 ml red cells and 150 ml supernatant (100 ml SAGM anticoagulant plus 50 ml plasma). During storage, potassium leaks from the red cells (intracellular potassium concentration is approximately 100 mmol.l−1) down a concentration gradient into all components of the supernatant (plasma and SAGM solution not just the plasma component). Supernatant levels of around 30 mmol.l−1 can be expected after 3 weeks storage. We transfused 2 units (748 ml) of SAGM blood preparation; this amounts to a total potassium load of 9 mmol. In a 42-kg cachectic woman with a calculated blood volume of 2520 ml (plasma volume of around 1500 ml) this potassium load has the potential to raise the potassium level by 6 mmol.l−1. But this would assume that complete intravascular retention takes place, whereas it is known that potassium usually equilibrates rapidly between the plasma and the extracellular fluid. The laboratory potassium level of 9.5 mmol.l−1 and the ECG changes confirm that there was indeed significant intravascular retention. We appreciate that modest transfusions of this nature are usually safe and in our article tried to explore other avenues for such a marked effect. Some suggestions given in our paper were: endogenous potassium release (tumour manipulation), reduced circulating volume and cardiac output, rapid pressure bag infusion, etc. However, the timing of the cardiac arrest and clinical presentation would suggest the transfusion must be implicated. Anaesthetists should not be lulled into a false sense of security simply because modest volumes of blood preparations are being transfused.

Prestorage Leucofiltration of Red Blood Cell Suspensions Modulates Storage Time-Dependent Suppression of in vitro Cytokine Release

Background: Leucocyte- and platelet-derived bioactive substances accumulated in blood preparations during procession and storage may play a role in side-effects, including immunosuppression, observed after transfusion. Leucodepletion by filtration may significantly reduce the amount of storage time-dependent accumulated substances. Therefore we studied the effect of pre-storage leucofiltration of red blood cell suspensions on stimulation of tumour necrosis factor-alpha (TNF-α) and interleukin-2 (IL-2) release in vitro. Materials and Methods: Nine units of CPDA-1 blood (whole blood suspended in citrate phosphate dextrose adenine-1), 9 units of pre-storage leucofiltered CPDA-1 blood, 9 units of SAGM blood (buffy coat-depleted red blood cells suspended in saline adenine glucose mannitol) and 9 units of pre-storage leucofiltered SAGM blood were stored for 35 days. Supernatants were collected from all units after storage for 1, 21 and 35 days. Assays of whole blood obtained from healthy volunteers with addition of supernatants from stored blood components were stimulated with lipopolysaccharide (LPS) or phytohemagglutinin A (PHA) and incubated for 24 h or 72 h. Control assays without addition of supernatants from stored blood components were run simultaneously. After incubation TNF-α and IL-2 concentrations were determined by ELISA methods in assay supernatants. Results: The stimulated TNF-α and IL-2 releases were significantly decreased by supernatants from CPDA-1 blood and SAGM blood in a storage time-dependent manner compared with release in control assays. Pre-storage leucofiltration of CPDA-1 blood improved TNF-α release significantly while leucofiltration of SAGM blood increased the release to concentrations higher than that observed in control assays. However, pre-storage leucofiltration had no effect on storage time-dependent decreased IL-2 release. Con-clusion: Pre-storage leucofiltration may improve certain in vitro immune responses impaired by supernatants from stored blood while others appear to be unaffected.

Leucocyte and platelet-derived bioactive substances in stored blood: effect of prestorage leucocyte filtration.

Adverse reactions to transfusion of allogeneic blood may depend on content of leucocytes and platelets and on storage-time of the erythrocyte suspensions. Therefore, we studied the efficacy of prestorage leucocyte reduction by filtration on total content and extracellular accumulation of histamine, eosinophil cationic protein (ECP), eosinophil protein X (EPX), myeloperoxidase (MPO), plasminogen activator inhibitor type-1 (PAI-1) and interleukin-6 (IL-6) in samples obtained from 5 units of SAGM blood, 7 units of plasma-reduced whole-blood and 6 units of whole-blood before and after filtration, respectively. In addition, we analysed supernatants from the same units after storage at +4 degrees C for 0, 21 and 35 d, respectively. The filtration was performed at room temperature within 2-4 h after donation. The substances were analysed by ELISA and RIA methods and we also analysed the donor plasma levels of the same bioactive substances. The total content of histamine, ECP, EPX, and MPO were 10-70-fold higher in all unfiltered erythrocyte products compared to donor plasma concentrations, while PAI-1 content was 15-20-fold higher only in plasma-reduced whole-blood and whole-blood. Prestorage leucocyte filtration significantly reduced the total histamine, ECP, EPX, MPO and PAI-1 content to levels similar to donor plasma levels in plasma-reduced whole-blood and whole-blood, while PAI-1 was still low in filtered SAGM blood. In addition, the levels of extracellular bioactive substances at d 0 after donation and filtration were within the range of concentrations in donor plasma, and there was no time-dependent accumulation during storage for 35 d at +4 degrees C. IL-6 was not detected in either plasma or samples obtained from the blood bags. These results suggest prestorage leucocyte filtration to deplete leucocyte contents to levels, which prevent the previously shown time-dependent accumulation of leucocyte derived bioactive substances in various erythrocyte suspensions. In addition, the PAI-1 results suggest leucocyte filters to reduce the obligatory platelet content in whole-blood products.

Bioactive substance accumulation and septic complications in a burn trauma patient: effect of perioperative blood transfusion?

Evidence has emerged that suggests adverse effects to perioperative homologous blood transfusion are related to the age of the blood products. Recently, time-dependent accumulation of bioactive substances in red cell suspensions, standard platelet concentrates and fresh frozen plasma during storage have been shown. The potential adverse effects of these bioactive substances were analysed in a burn trauma patient. A patient with 40 per cent second and third degree burn trauma without other injuries underwent a two-step transplantation operation. Samples for analyses of histamine, eosinophil cationic protein (ECP), eosinophil protein X (EPX), neutrophil myeloperoxidase (MPO) and interleukin 6 (IL-6) were drawn frequently from the patient before, during and after the operations, and from all transfused red cell, platelet and fresh frozen plasma units. Urine was sampled every hour during the first operation for analyses of ECP and EPX excretion. All analyses were performed by ELISA and RIA methods, and results compared to patient outcome. The patient received a total of 48 and 8 SAGM blood, 6 and O platelet and 12 and 4 fresh frozen plasma units at the two operations, respectively. Transfused products contained a total of 64.54 nmol and 17.50 nmol histamine, 115518 ng and 25764 ng ECP, 174457 ng and 38770 ng EPX, 6950915 ng and 1505125 ng MPO, and 14740 pg and 5600 pg IL-6 at the two operations, respectively. The accumulation of the substances in patient plasma correlated to postoperative septic reactions, without any disclosure of bacteraemia after the first operation, while the accumulation at the second operation correlated to the septic reaction and Pseudomonas aeruginosa infection. Timedependent accumulation of bioactive substances in blood products during storage may be related to the development of post-transfusion adverse effects.

Time-dependent histamine release from stored human blood products

Perioperative transfusion of whole blood has been shown to amplify trauma-induced immunosuppression, which could be attenuated by perioperative administration of histamine2 receptor antagonists. Supernatants from different blood products were, therefore, analysed for histamine content during storage. Whole blood (six units), plasma-reduced whole blood (six units), and plasma- and buffy coat-reduced (saline-adenine-glucose-mannitol) (SAGM) blood (six units) from unpaid healthy donors were stored in the blood bank for 35 days at 4 degrees C. Plasma histamine and total cell-bound histamine content at donation, and histamine concentration in samples drawn from the units on days 0, 2, 5, 9, 14, 21, 28 and 35 were analysed with an enzyme-linked immunosorbent assay. Median plasma histamine concentration was 4.8 (range 1.9-14.3) nmol/l (n = 18). Median total cell-bound histamine content was 417.0 (range 176.0-910.0) nmol/l in whole blood and 475.0 (range 360.0-1560.0) nmol/l in plasma-reduced whole blood, while it was undetectable in SAGM blood. Spontaneous histamine release increased in a time-dependent manner from a median of 6.7 (range 2.2-17.4) nmol/l at the time of storage to 175.0 (range 33.0-485.0) nmol/l at day 35 in whole blood, from 18.8 (range 8.2-38.5) to 328.5 (range 224.0-1137.0) nmol/l in plasma-reduced whole blood, and from 0.5 (range 0.5-1.5) to 2.2 (range 1.4-6.9) nmol/l in SAGM blood. These results show spontaneous histamine release during storage of human blood products which contain full amount of leucocytes, and this may play a significant role in detrimental effects of blood transfusion.

Autotransfusion after open heart surgery: quality of shed mediastinal blood compared to banked blood

The need to conserve a patient's own blood and avoid homologous transfusion is now well recognized. Therefore, techniques designed to reduce requirements for homologous blood transfusions have been developed. One of the methods is autotransfusion of shed mediastinal blood after open-heart surgery. The objectives of the present study were to investigate osmotic fragility and oxygen transport capacity of shed mediastinal blood compared to patient blood and stored packed red blood cells (SAGM). Shed mediastinal blood from ten consecutive patients undergoing elective cardiac surgery (coronary bypass grafting) was studied and compared to patient blood, 10 units of 3 weeks old and 10 units of 5 weeks old stored packed red blood cells (SAGM). Oxygen transport capacity was investigated by calculation of p50 for oxygen by use of the oxygen status algorithm (OSA 2.0) programme and measurement of 2,3-diphosphoglycerate (2,3-DPG) concentrations. The osmotic fragility was determined using increasing concentrations of saline. 2,3-DPG concentrations in shed mediastinal blood (5.3 mikromol/ml erythrocyte) were within the range measured in patient blood, but significantly higher than SAGM blood (P < 0.001). P50 for oxygen (3.5 kPa) in shed mediastinal blood was not significantly different compared to patient blood, but significantly higher (P < 0.01) compared with stored SAGM blood. The osmotic fragility in shed mediastinal blood was not significantly different compared to patient blood, but significantly lower (P < 0.001) than the osmotic fragility in stored SAGM blood. This suggests that red cells saved from shed mediastinal blood have better oxygen transport capacity and may have longer survival compared to stored blood.

Concern over safety of SAGM blood

Concern over safety of SAGM blood

In vitro Haemolysis of SAG-M and CPD-A Blood Units

In vitro Haemolysis of SAG-M and CPD-A Blood Units

In vitro Haemolysis of SAG-M and CPD-A Blood Units

In vitro Haemolysis of SAG-M and CPD-A Blood Units