The purification of red cells for transfusion by freeze-preservation and washing. V. Red cell recovery and residual leukocytes after freeze- preservation with high concentrations of glycerol and washing in various systems

Red blood cells freeze-preserved with 40% W/V glycerol in an ionic or low ionic medium at −80 C were washed in one of four commercially available systems: the Haemonetics Blood Processor 15 using a continuous-flow centrifugation procedure, the Fenwal Elutramatic using a continuous-flow centrifugation procedure, the IBM Blood Processor using an automated serial centrifugation procedure, and the Huggins Cytoglomerator using a dilution/agglomeration procedure. Determinations of red blood cell recovery and leukocyte and platelet removal were made for each of these groups. The dilution/agglomeration procedure produced lower red blood cell recovery values and poorer leukocyte and platelet removal than did any of the three wash systems using sodium chloride solutions. The values obtained with the systems using sodium chloride solutions were slightly but significantly different.     

Paroxysmal nocturnal hemoglobinuria and the transfusion of washed red cells. A myth revisited

Paroxysmal nocturnal hemoglobinuria (PNH) is an uncommon, acquired clonal stem cell disorder primarily affecting red cells that have an abnormal sensitivity to complement lysis. Since 1948, the use of saline-washed red cells (WRBCs) has been advocated to minimize hemolysis after transfusion to patients with PNH. Thirty-eight years of experience (1950 through 1987) with patients who had PNH were reviewed. Twenty-three patients with a positive Ham's test had been transfused with 556 blood components, including 431 RBC products: 94 units of whole blood, 208 units of packed RBCs, 80 units of white cell-poor RBCs, 38 units of WRBCs, 5 units of frozen RBCs, and 6 units of intraoperatively salvaged RBCs. Only one documented episode of posttransfusion hemolysis related to the underlying diagnosis of PNH was found, and it was associated with the transfusion of a unit of type O whole blood to an AB-positive individual. This unit contained ABO-incompatible plasma; this case was similar to one in an earlier report from which originated the recommendation for using WRBCs. The posttransfusion increment in hemoglobin concentration in patients receiving ABO-identical packed RBCs was comparable to that in patients receiving frozen or washed RBCs. These findings indicate that the use of WRBCs is unnecessary and that patients with PNH should be transfused with group-specific blood and blood products.     

Transmission of human T-lymphotrophic virus type I infection to a neonatal infant by transfusion of washed and irradiated red cells

Human T-lymphotropic virus type I and/or II (HTLV-I/II) may be transmitted by the transfusion of blood and blood components. Several factors are critical to the efficiency of transmission. These include the number of contaminating white cells, the component volume, and the age of the component. After look-back notification, there was an investigation of the HTLV-I/II serostatus of three patients who had received blood from a donor now found to be positive for HTLV-I antibodies by enzyme immunoassay and Western blot. The donor red cell unit was group O negative and cytomegalovirus antibody negative; it was washed and irradiated at 2800 cGy and aliquoted into six small-volume transfusions for four neonatal infants. Three of the four patients were available for testing 3.5 years after their exposure. The fourth neonatal infant died on Day 11 of life. The three tested infants received 14, 14, and 44 mL of component, respectively. HTLV-I seroconversion was documented by enzyme immunoassay and Western blot (p19, p24) and occurred only in the patient receiving 44 mL. On the basis of quality control data, it is estimated that the affected infant received 8 × 10(7) white cells in the 44-mL aliquot. Washing and irradiation will not necessarily eliminate HTLV-I transmission.     

Application of frozen plasma to red cell consumption ratios as a possible indicator for audit of use of frozen plasma in hospital clinical transfusion practice

The consumption of frozen plasma expressed as a function of consumption of red cells has been used to compare frozen plasma transfusion practices in various countries. This principle has been applied to consumption of frozen plasma in individual hospitals as a potential indicator of the desirability of audit of transfusion practice at the individual hospital level.Wide variation in the relative consumption of frozen plasma has been found both between groups of hospitals determined by size and academic affiliation and within these hospital groupings. Further, there is a close correlation between consumption of frozen plasma in relation to red cell consumption and the annual consumption of frozen plasma in relation to active (acute) treatment beds.It is suggested that high frozen plasma:red cell transfusion ratios may serve as an indicator of a need for audit of frozen plasma transfusion.     

White cell-reduced red cells prepared by filtration: a critical evaluation of current filters and methods for counting residual white cells

White cell (WBC) reduction, red cell (RBC) recovery, and filtration time were determined in 1-day-old standard and buffy coat-depleted RBCs filtered in the laboratory through six commercial filters for WBC reduction. Residual WBCs were counted with a Burker chamber (BC), with a Nageotte chamber (NC), and by flow cytometry (FC). Results show that BC counts were 0 in several cases in which WBCs were detected with NC and FC, which indicated that the traditional BC method is too insensitive in use with currently available filters. Calibration curves performed by FC and with NC with samples containing known concentrations of WBCs from 1000 to 1 per microL showed that both FC and NC detected, on average, 67 percent of WBCs present in the samples (efficiency). However, the efficiency of FC showed small variability (61-70%) at different WBC levels, whereas the variability with NC was large (39-91%). This greater variability prevented the correction of NC counts by using a single factor and indicated difficulty in NC standardization. Therefore, because our main aim was to compare different filters rather than to define absolute levels of WBC contamination, uncorrected FC and NC counts were chosen to be reported. True WBC counts per unit should not exceed values that can be obtained by dividing uncorrected counts by the lowest efficiencies (61% for FC and 39% for NC). Uncorrected NC and FC counts were below 2 × 10(6) per unit in all units processed through three of the filters and below 5 × 10(6) per unit in all units processed through the other three.(ABSTRACT TRUNCATED AT 250 WORDS)     

The use of a blood conservation device to reduce red blood cell transfusion requirements: a before and after study

Introduction  

Anaemia and the associated need for packed red blood cell (PRBC) transfusions are common in patients admitted to the intensive care unit (ICU). Among many causes, blood losses from repeated diagnostic tests are contributory.     

The preparation of leukocyte-poor red cells for transfusion by a simple cost-effective technique

A modification of the microaggregate filtration technique for the production of leukocyte-poor red cell concentrate is described. The modified technique, termed 'spin, cool, and filter', removed leukocytes more efficiently from relatively fresh blood than the original microaggregate filtration procedure. The residual leukocyte content of 4- to 9-day-old units processed by the modified procedure averaged less than 0.4 X 10(9) white cells per unit. The microaggregate filtration technique, supplemented by buffy-coat exclusion, removed more than 93 percent of the units' white cells. These red cell products were prepared easily and did not elicit febrile reactions in highly susceptible patients.     

The manual preparation of leukocyte-poor red cells for transfusion by a new filter

Standard and buffy-coat-depleted red cell concentrates were filtered through a new cellulose acetate filter with a manual procedure. No residual leukocytes could be detected in 19 of the 21 filtered units. Extracellular hemoglobin after red cell filtration and concentration never exceeded 10 mg per unit. No transfusion reaction occurred in 10 recipients with potent lymphocytotoxic antibodies in their serum and histories of febrile transfusion reactions. This filter allows the preparation of leukocyte-poor red cell units with a safe, simple, and effective manual procedure.     

In vitro measurement of P50--the pH correction, and use of frozen red blood cells as controls.

The Unicam Spectrophotometer, the IL 282 CO-Oximeter, and the Hemoscan were used to measure the in vitro P50 using 1, 20--45, and 50% hematocrit values of red blood cells suspended in autologous plasma or phosphate buffered saline at pH of 6.7--7.7. The relation between pH and P50 was a linear correlation for washed red blood cells by all three methods whereas the relation between pH and P50 was a log correlation for red blood cells in plasma. Multiple units of red blood cells with low 2,3 diphosphoglycerate (2,3 DPG) and increased affinity for oxygen, with normal 2,3 DPG and normal affinity for oxygen, and above normal 2,3 DPG and decreased affinity for oxygen, were frozen in 10 ml aliquots with 40% w/v glycerol and maintained at -80 degrees C. One tube of each level of 2.3 DPG was thawed, washed, and suspended in buffer daily to yield P50 values of 17.5 +/- 0.67, 29.7 +/- 0.92, or 41.2 +/- 0.72 torr at pH 7.2 in order to have a control of both instrumentation and personnel.     

Prevention of adverse reactions to blood transfusion by the administration of saline-washed red blood cells

We prospectively compared the incidence of complications following saline-washed versus packed red blood cell transfusions, to determine whether routine use of washed red blood cells could reduce significantly the incidence of transfusion reactions. Clinical reports of reactions were evaluated carefully to confirm whether the reaction was caused by transfusion. In 3,799 washed red blood cell transfusions, there were eight confirmed reactions (0.21%). Of 6,359 packed red blood cell transfusions, 31 reactions occurred (0.49%). The difference in incidence of confirmed complications was statistically significant (p less than 0.03). Administration of washed red blood cells to all patients requiring transfusions can thus be seen to reduce significantly the incidence of adverse reactions. This is likely the result of the removal of leukocytes and plasma achieved by the washing process. The increased safety of washed red blood cells must be weighed against their extra expense to determine their cost-effectiveness in transfusion therapy.     

Analysis of a linear peristaltic infusion device for the transfusion of red cells to pediatric patients

A linear peristaltic infusion device was evaluated for red cell (RBC) transfusion in the pediatric and neonatal setting. CPDA-1 RBC units (n = 24) divided into six groups of 4 units each underwent simulated transfusion. Blood was infused by using manufacturer-provided administration sets with either a 21-gauge needle or a 24-gauge catheter. Filters were used in two groups to evaluate the effect of negative pressure on filter function. Two groups of RBCs less than 1 week old were washed, irradiated, and infused at 5 mL per hour, by using a standard administration set, or at 10 mL per hour, by using a syringe set. Four-week-old RBCs (washed and irradiated, irradiated and filtered, filtered only, or unmanipulated) were infused at 100 mL per hour. Paired samples from 0 and 2 hours before and after infusion were analyzed for hemoglobin, hematocrit, RBC count, plasma hemoglobin, lactate dehydrogenase, potassium, alanine aminotransferase, and aspartate aminotransferase. Hausser and Nageotte hemocytometers were used to perform white cell (WBC) counts when a filter was used. By analysis of variance and percentage of change, data from 0 and 2 hours before and after infusion were compared. No clinically or statistically significant differences were seen for hemoglobin, hematocrit, or RBC count. The difference in preinfusion and postinfusion plasma hemoglobin levels in washed RBCs at 2 hours was statistically but not clinically significant (14.5 +/− 6.8 vs. 19.3 +/− 7.1 mg/dL). No clinically significant differences were noted for the remaining analytes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Therapeutic effectiveness and safety of outdated human red blood cells rejuvenated to improve oxygen transport function, frozen for about 1.5 years at 80 C, washed, and stored at 4 C for 24 hours prior to rapid infusion

After storage at 4 C for 20 to 28 days, red blood cells were biochemically modified to improve their oxygen transport function which had deteriorated during liquid storage. The solution used for rejuvenation contained pyruvate, inosine, glucose, phosphate, and adenine (PIGPA Solution B). The rejuvenated red blood cells were frozen with 40% W/V glycerol in a polyolefin plastic bag and were stored in the frozen state for about 1.5 years at −80 C. After thawing and washing the red blood cells were stored at 4 C in a sodium chloride- glucose-phosphate solution for 24 hours before transfusion. A pool of four to ten units was rapidly transfused to each of 14 elderly anemic recipients, 11 of whom had cardiopulmonary insufficiency. Recovery of the red blood cells after the freeze-thaw process was about 97 per cent, and after the freeze-thaw-wash process about 90 per cent. The 24− hour posttransfusion survival values were about 75 per cent, and the long-term survival values were about 85 days depending on the disease state of the recipient. The red blood cells had 1.5 times normal 2.3- DPG levels and a decreased affinity for oxygen at the time of transfusion and were able to delivery oxygen at high oxygen tension immediately after the rapid infusion of pools of from four to ten units through a 40-or 170-micron filter. Plasma hemoglobin levels were consistent with extravascular sequestration of nonviable red blood cells, and uric acid levels were not increased during the immediate 24− hour posttransfusion period.     

The use of immunoabsorbents for the purification of mycobacterial antigens.

Affinity chromatography using immunoabsorbents prepared from goat antisera has been applied to the purificaton of antigens from culture filtrates of Mycobacterium tuberculosis. The optimum conditions for the preparation and use of these immunoabsorbents have been determined. Urea has been studied as an eluant, and it has been found that antigen can be eluted with 4M urea at pH 9 with satisfactory yields, minimal loss of antigenicity, and minimal disruption of the immunoabsorbent.     

Therapeutic effectiveness and safety of outdated human red blood cells rejuvenated to restore oxygen transport function to normal, frozen for 3 to 4 years at −80 C, washed, and stored at 4 C for 24 hours prior to rapid infusion

Human red blood cell concentrates with hematocrit values of 75 V% were prepared from citrate-phosphate-dextrose (CPD) blood, stored at 4 C for 20 to 28 days, and biochemically modified with a solution containing pyruvate, inosine, glucose, phosphate, and adenine (PIGPA Solution A). The rejuvenated red blood cells were frozen with 40% W/V glycerol in a polyolefin plastic bag and were stored at ?80 C. After three to four years of frozen storage, the units were thawed, washed, and stored at 4 C in a sodium chloride-glucose-phosphate solution for 24 hours prior to transfusion. Red blood cell recovery was 97 per cent after thawing and 90 per cent after washing. An automated differential agglutination procedure (ADA) showed 24-hour survival values of about 80 per cent, and long-term survival values of about 85 days depending on the disease state of the recipient. The red blood cells had normal affinity for oxygen on the day of transfusion. Plasma hemoglobin levels measured immediately after transfusion indicated extravascular removal of nonviable donor red blood cells. There was no increase in the uric acid level during the 24-hour posttransfusion period. A pool of three to ten units of rejuvenated washed previously frozen red blood cells was transfused rapidly to each of 19 anemic elderly patients. The red blood cells which had normal oxygen delivery capacity immediately upon transfusion increased the recipient's red blood cell mass and produced no untoward effects.     

The 24-hour posttransfusion survival, oxygen transport function, and residual hemolysis of human outdated-rejuvenated red cell concentrates after washing and storage at 4 degrees C for 24 to 72 hours

Red cell concentrates with hematocrit values of 80 +/- 5 percent were stored at 4 degrees C either in citrate-phosphate-dextrose for 22 to 28 days or in citrate-phosphate-dextrose-adenine-one for 35 to 39 days. After storage, the red cells had reduced 2,3-diphosphoglycerate and adenosine triphosphate levels, and biochemical treatment with a solution called PIPA was used to restore these levels. The red cells were not preserved further, but instead were washed after rejuvenation with an unbuffered sodium chloride-glucose solution, pH 5.0, for in vivo studies, and with a buffered sodium chloride-glucose-phosphate solution, pH 6.8, for comparative in vitro studies. The red cells were stored in the wash solution at 4 degrees C for 72 hours after washing. Red cell recovery after washing was about 95 percent. Twenty-four-hour posttransfusion survival value was about 80 percent, and the index of therapeutic effectiveness was greater than 75 percent. These biochemically modified washed red cells exhibited higher than normal P50 values, even after 3 days of postwash storage at 4 degrees C. The units that were washed with the sodium chloride-glucose solution with a pH of 5.0 exhibited a greater degree of hemolysis after 3 days of postwash storage at 4 degrees C than did the units that were washed with the sodium chloride-glucose-phosphate solution with a pH of 6.8.(ABSTRACT TRUNCATED AT 250 WORDS)