Post-transfusion Recovery Research Articles

Studies with nonradioisotopic sodium chromate

A recently developed nonradioisotopic 52Cr technique was used to measure either red cell volume or posttransfusion recovery of stored red cells. The experimental method uses Zeeman electrothermal atomic absorption spectrophotometry to measure red cell chromium. Results from the 52Cr method were compared with those from 51Cr single-label and 125I-albumin/51Cr double-label procedures using 49-day AS-1 red cell concentrates drawn and prepared according to standard procedures. In the first group of five donors, red cell volume was estimated concurrently with both 52Cr-labeled fresh red cells and 125I-albumin. The latter measured plasma volume from which red cell volume was estimated on the basis of the hematocrit (125I red cell volume). 51Cr-labeled stored red cells were transfused to measure posttransfusion recoveries. The correlation between 52Cr and 125I red cell volumes was significant (r = 0.68, p less than 0.01), and, in this group, the differences were not significant (p less than 0.05). Twenty-four-hour posttransfusion recoveries of 51Cr-labeled stored red cells averaged 66 +/- 5 percent when measured with the 125I/51Cr technique and 69 +/- 8 percent when measured with the 52Cr/51Cr method. In the second group of five donors, red cell volume was estimated by the 125I-albumin technique, and the posttransfusion recovery of stored red cells was quantitated by 51Cr- and 52Cr-labeled stored cells simultaneously. In this group, posttransfusion recoveries with 125I/51Cr averaged 73 +/- 7 percent; with 125I/52Cr, they averaged 75 +/- 10 percent. Using the single-label method of calculation, recoveries averaged 76 +/- 7 and 75 +/- 10 percent for the 51Cr and 52Cr methods, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of 99mTechnetium/51Chromium Post-Transfusion Recovery of Red Cells Stored in Saline, Adenine, Glucose, Mannitol for 42 Days

An in vivo and in vitro study of 42-day saline, adenine, glucose, mannitol (SAGM) red cells (Terumo Corporation, Elkton, Md.) was conducted using 20 volunteers to document in vivo efficacy and analyze the validity of the 99mTc/51Cr technique. In vivo autologous post-transfusion recovery was measured by a double-label procedure involving the use of 99mTc-labeled freshly drawn red cells to quantify recipient blood volume and 51Cr to document both posttransfusion 24-hour recovery (PTR) and red cell survival. As an internal control, 5 of the 20 donors studied had red cell volumes estimated by the 125I-albumin (RISA) plasma volume technique on a separate occasion. For comparison, PTR was also calculated according to the single-label 51Cr protocol in which recipient blood volume was estimated by extrapolation from circulating 51Cr-labeled red cell activity 5-15 min after infusion. After 42 days of storage, PTR averaged 78±6% with the double-label 99mTc/51Cr technique and 81±5% with the single-label (51Cr alone), confirming a small difference between the two techniques. Correlation between the two techniques was high, r = 0.81, though the average 3% difference between them was significant (p<0.05). Red cell volumes of the 5 donors measured by both the 99mTc-red cell method and the 125I-albumin method exhibited excellent correlation, r = 0.96, and averaged 1,961 ±420 ml and 2,048±381 ml, respectively. Standard in vitro parameters of SAGM red cell concentrates were well preserved with hemolysis of 0.5±0.4% and red cell ATP 69±13% of the initial values after 42 days of storage. The double-label, 99mTc/51Cr, post-transfusion recoveries exhibited a slightly better correlation with post-storage ATP when compared to single-label, 51Cr, recoveries: r = 0.705 versus r = 0.644, respectively. The results confirmed acceptable 42-day storage parameters for modified SAGM red cells.

Recovery and survival in vivo of platelet concentrates prepared with prostaglandin E1.

In follow-up to previous studies showing that stimulators of adenylate cyclase inhibit the activation of platelets in platelet concentrates (PC), the posttransfusion recovery and survival of autologous platelets prepared and stored after the addition of prostaglandin E1 (PGE1) to platelet-rich plasma at a concentration of 1.3 X 10(-8) M were investigated. Six normal subjects were studied on the two occasions, using PC stored with and without PGE1 and radiolabeling with 51Cr. The mean recovery of platelets prepared with PGE1 (35.2 +/- 8.1%) was significantly lower (p less than 0.013) than that of routinely prepared platelet concentrates (46.3 +/- 9.4%). The mean life-spans of platelets prepared with and without PGE1 were 7.1 +/- 0.4 and 6.2 +/- 1.0 days, respectively (p = NS). Despite its ability to inhibit the activation of platelets during concentration and storage, prostaglandin E1 appears to reduce posttransfusion recovery of platelets significantly in this experimental model and cannot be recommended at this time as an adjunct for PC preparation.

Evaluation of Four Methods for Platelet Compatibility Testing

Four platelet compatibility assays were performed on serum and platelet or lymphocyte samples from 38 closely HLA-matched donor/recipient pairs involved in 55 single-donor platelet transfusions. The 22 patients studied were refractory to transfusions of pooled random-donor platelets. Of the four assays (platelet suspension immunofluorescence, PSIFT; 51Cr release; microlymphocytotoxicity; and a monoclonal anti-IgG assay, MAIA), the MAIA was most predictive of platelet transfusion outcome (predictability, 74% for one-hour posttransfusion platelet recovery and 76% for 24-hour recovery). The only other assay to reach statistical significance was the PSIFT (63% predictability for one-hour posttransfusion recovery). The degree of HLA compatibility between donor and recipient (exact matches v those utilizing cross-reactive associations) was unrelated to the ability of the MAIA to predict transfusion results. The MAIA may be capable of differentiating HLA antibodies, ABO antibodies, and platelet-specific antibodies responsible for failure of HLA-matched and selectively mismatched single-donor platelet transfusions.

Evaluation of four methods for platelet compatibility testing

Four platelet compatibility assays were performed on serum and platelet or lymphocyte samples from 38 closely HLA-matched donor/recipient pairs involved in 55 single-donor platelet transfusions. The 22 patients studied were refractory to transfusions of pooled random-donor platelets. Of the four assays (platelet suspension immunofluorescence, PSIFT; 51Cr release; microlymphocytotoxicity; and a monoclonal anti-IgG assay, MAIA), the MAIA was most predictive of platelet transfusion outcome (predictability, 74% for one-hour posttransfusion platelet recovery and 76% for 24-hour recovery). The only other assay to reach statistical significance was the PSIFT (63% predictability for one-hour posttransfusion recovery). The degree of HLA compatibility between donor and recipient (exact matches v those utilizing cross-reactive associations) was unrelated to the ability of the MAIA to predict transfusion results. The MAIA may be capable of differentiating HLA antibodies, ABO antibodies, and platelet-specific antibodies responsible for failure of HLA-matched and selectively mismatched single-donor platelet transfusions.

Studies of the minimum temperature at which human platelets can be stored with full maintenance of viability.

Platelet concentrates from normal donors were stored for 3 days under identical conditions except for the temperature of storage, which was maintained at 21 +/- 0.5, 19.5 +/- 0.5, or 18 +/- 0.5 degrees C. Immediate posttransfusion recovery of the stored platelets determined by 51Cr labeling averaged 47, 47, and 48 percent after storage at 21, 19.5, and 18 degrees C, respectively (differences not significant). Mean life span of the transfused platelets, however, was 8.12, 5.21, and 1.85 days at 21, 19.5, and 18 degrees C, respectively. The difference between mean life span following storage at 21 degrees C was significantly different from that after storage at 18 degrees C (p less than 0.03). Reduction in viability after storage at the lower temperature correlated with the reduction in the number of discoid platelets. These findings indicate that platelet viability is compromised after storage for 3 days at 18 degrees C and, possibly, at 19.5 degrees C, and illustrate the need for quality control of temperature in short-term platelet storage.

Single donor platelet transfusion.

Cytopheresis equipment from a number of manufacturers now permits the rapid, safe, and eflicient procurement of functionally normal platelets from Single donors. There has been an extensive proliferation of these blood cell separators and virtually all large blood collection centers and most large hospitals have at least one blood cell Separator in Operation. Transfusions from Single donors selected by HLA typing are a wellaccepted feature of the management of alloimmunized patients [1, 2]. In addition, occasional centers located at a distance from regional blood transfusion centers utilize nonmatched transfusions from Single donors as a major part of their “regular” platelet supply. Because most Single donor transfusions are not obtained from “closed systems” however, these collections cannot be stored for more than 24 h, making it more difficult to utilize such produets rationally as a form of long-term platelet inventory. Furthermore, the availability of newer plastic bags now allows storage of platelet concentrates at ambient temperatures with preservation of normal posttransfusion recovery after 7 days of storage [3]. Thus, there should be few such “geographic peculiarities” which necessitate the widespread use of Single donor platelets as “random donor platelets” in the future.

Use of Adsol preservation solution for prolonged storage of low viscosity AS-1 red blood cells.

A new red cell preservation solution is described in which the red cells may be stored for 49 d with greater than 75% mean post-transfusion recovery. Blood is drawn into Anticoagulant Citrate Phosphate Dextrose Solution, centrifuged, the plasma removed and the cells resuspended in 100 ml of a solution containing saline, adenine, dextrose and mannitol (Adsol Preservation Solution, Fenwal Laboratories, Deerfield, Illinois). The final product, AS-1 Red Blood Cells, has a haematocrit of approximately 0.60 and flow properties that are similar to those of whole blood. After storage, red cell haemolysis is minimal and erythrocyte adenosine triphosphate is well preserved. This study documents that red cells may be stored in a protein-poor electrolyte medium for periods of 49 d with good post-transfusion survival.

急性白血病における大量か粒球輸血の回収率を決定する因子に関する研究

The results of 68 leukocyte cross-matching procedure have been assessed in order to select compatible donors for 19 febrile granulocytopenic patients with acute leukemia.It was revealed that there was positive relationship between precounts of patients' own leukocytes and granulocyte recovery rates. This phenomenon suggested that leukocytes of marginal pool were markedly depleted and most part of transfused granulocytes might marginate along the vessel walls in leukopenic patients.This study confirms the clinical value of lymphocytotoxicity test (LCT) in predicting the post-transfusion recovery rate of single donor granulocytes. There was a good agreement between the LCT croos-match and granulocyte recovery. In general, compatibility in the granulocytotoxicity (GCT) has not any significant value in granulocyte recovery rates.

Effect of HLA Bw4/Bw6 compatibility on platelet transfusion responses of refractory thrombocytopenic patients

Platelet transfusions from donors matched for cross-reactive antigens have been shown to be effective in providing hemostasis in alloimmunized thrombocytopenic patients. A significant number of these transfusions, however, fail to provide posttransfusion platelet recoveries. We investigated incompatibility in the Bw4/bw6 system as a possible explanation for these failures. The Bw4/Bw6 system is a biallelic antigen system closely associated with HLA-B. HLA-B locus antigens that are cross-reactive frequently differ in their Bw4/Bw6 specificity. Posttransfusion platelet recoveries from 21 alloimmunized thrombocytopenic patients homozygous for Bw4 or Bw6 and transfused with both Bw4/Bw6 compatible and incompatible platelets were analyzed. The mean 1-hr posttransfusion recovery was 84% following Bw4/Bw6-compatible platelets versus 52% with Bw4/Bw6-incompatible platelets (p less than 0.02). Twenty-four hours following transfusion, mean recoveries were 44% and 24%, respectively, (p less than 0.01). A subgroup of 8 patients (38%) was identified who had markedly lower responses following Bw4/Bw6- incompatible transfusions as compared to Bw4/Bw6-compatible transfusions (mean recoveries: 1 hr--compatible 100%, incompatible 27%, p less than 0.001; 24 hr--compatible 45%, incompatible 7%, p less than 0.01). These data suggest that the Bw4/Bw6 antigen system has clinical significance for some patients requiring platelet transfusion therapy and, when appropriate, should be considered in the selection of donors.

Effect of HLA Bw4/Bw6 Compatibility on Platelet Transfusion Responses of Refractory Thrombocytopenic Patients

Platelet transfusions from donors matched for cross-reactive antigens have been shown to be effective in providing hemostasis in alloimmunized thrombocytopenic patients. A significant number of these transfusions, however, fail to provide posttransfusion platelet recoveries. We investigated incompatibility in the Bw4/bw6 system as a possible explanation for these failures. The Bw4/Bw6 system is a biallelic antigen system closely associated with HLA-B. HLA-B locus antigens that are cross-reactive frequently differ in their Bw4/Bw6 specificity. Posttransfusion platelet recoveries from 21 alloimmunized thrombocytopenic patients homozygous for Bw4 or Bw6 and transfused with both Bw4/Bw6 compatible and incompatible platelets were analyzed. The mean 1-hr posttransfusion recovery was 84% following Bw4/Bw6-compatible platelets versus 52% with Bw4/Bw6-incompatible platelets (p less than 0.02). Twenty-four hours following transfusion, mean recoveries were 44% and 24%, respectively, (p less than 0.01). A subgroup of 8 patients (38%) was identified who had markedly lower responses following Bw4/Bw6-incompatible transfusions as compared to Bw4/Bw6-compatible transfusions (mean recoveries: 1 hr--compatible 100%, incompatible 27%, p less than 0.001; 24 hr--compatible 45%, incompatible 7%, p less than 0.01). These data suggest that the Bw4/Bw6 antigen system has clinical significance for some patients requiring platelet transfusion therapy and, when appropriate, should be considered in the selection of donors.

Human sickle erythrocytes: survival in chimpanzees.

The survival characteristics of human sickle (SS) erythrocytes (RBCs) transfused to intact chimpanzees were determined. The mean post-transfusion recovery of 51Cr-labelled SS RBCs in four chimpanzees was 30.5% +/- 15.2 SD, and the half-life survival was 4.2 h +/- 0.8 SD. The recovery of control (hemoglobin AA) human red cells in five chimpanzees was complete and their mean intravascular T 1/2 was 22.3 h. Shorter survival of sickle erythrocytes was also shown by transfusing chimpanzees with mixtures of human cells such as 51Cr AA RBCs and 59Fe SS RBCs, or 51Cr SS RBCs and non-labelled fetal (cord blood) erythrocytes. The difference in survival of AA and SS RBCs resembles that in human recipients and was probably caused by sickling of SS cells in the chimpanzee circulation. These primate animals could, therefore, be used as a model for in vivo studies of sickle cell disease.

Viability and Function of Stored Sickle Erythrocytes

Functional and metabolic characteristics of fresh and three-week stored erythrocytes from patients with sickle cell anemia were compared. The storage-related changes in ATP, 2,3-DPG, and P50 in sickle erythrocytes were similar to those in control (HbA) red blood cells. After storage in CPD, sickle erythrocytes maintained significantly higher levels of 2,3-DPG (mean 2.20 +/- 0.73 mM/ml RBC) than did control cells (mean 0.36 +/- 0.13 mM/ml RBC). The posttransfusion recovery and survival of stored SS erythrocytes in autologous recipients and in an animal test system were at least as good as those before storage. Tolerance of the storage lesion by sickle erythrocytes is probably related to their young mean cell age. These results also suggest that the option of autotransfusion should be explored for selected patients with sickle cell disease in special clinical settings.

In Vitro Function and Post-Transfusion Survival of Granulocytes Collected by Continuous-Flow Centrifugation and by Filtration Leukapheresis

The function of granulocytes collected by continuous-flow centrifugation (CFC) and by filtration leukapheresis (FL) was studied in vitro, and the post-transfusion recovery and intravascular survival of these cells was studied by autologous transfusion in normal donors. Granulocytes collected by both FL and CFC leukapheresis (CFCL) functioned normally in the quantitative nitroblue tetrazolium, oxygen consumption, and chemotaxis assays. Bacterial killing was slightly but consistently decreased in FL but not CFCL granulocytes. The post-transfusion recovery of control granulocytes collected by ordinary phlebotomy averaged 52% in eight transfusions, compared with 34% for six CFCL granulocyte concentrates and 16% for six FL concentrates. The intravascular half-times were 3.8 hr for phlebotomy and 3.0 hr for CFCL granulocytes. FL granulocytes had survival curves which were nonlinear and a single half-life could not be calculated. The average half-time 30 min after transfusion was 1.3 hr, and 3 hr after transfusion it was 2.6 hr. Granulocytes collected by FL had a mild impairment of bacterial killing, decreased post-transfusion recovery, and altered intravascular kinetics. None of these abnormalities was found in granulocytes collected by CFCL.

In vitro function and post-transfusion survival of granulocytes collected by continuous-flow centrifugation and by filtration leukapheresis

The function of granulocytes collected by continuous-flow centrifugation (CFC) and by filtration leukapheresis (FL) was studied in vitro, and the post-transfusion recovery and intravascular survival of these cells was studied by autologous transfusion in normal donors. Granulocytes collected by both FL and CFC leukapheresis (CFCL) functioned normally in the quantitative nitroblue tetrazolium, oxygen consumption, and chemotaxis assays. Bacterial killing was slightly but consistently decreased in FL but not CFCL granulocytes. The post- transfusion recovery of control granulocytes collected by ordinary phlebotomy averaged 52% in eight transfusions, compared with 34% for six CFCL granulocyte concentrates and 16% for six FL concentrates. The intravascular half-times were 3.8 hr for phlebotomy and 3.0 hr for CFCL granulocytes. FL granulocytes had survival curves which were nonlinear and a single half-life could not be calculated. The average half-time 30 min after transfusion was 1.3 hr, and 3 hr after transfusion it was 2.6 hr. Granulocytes collected by FL had a mild impairment of bacterial killing, decreased post-transfusion recovery, and altered intravascular kinetics. None of these abnormalities was found in granulocytes collected by CFCL.

The Increased Effectiveness of Platelet Concentrates Prepared in Acidified Plasma

AbstractPlatelet concentrates prepared in acidified plasma (pH 6.5-6.7) are superior to concentrates prepared by standard methods, and are 80-90 per cent as effective as platelet rich plasma (PRP). The use of excess citric acid to acidify plasma promotes resuspension of the concentrate by eliminating clumping, which is a major factor in the decreased effectiveness of standard concentrates. Analysis of posttransfusion recovery and survival of platelets reveals no evidence of platelet injury in an acid medium.Acidification of PRP inhibits the aggregation of platelets by adenosine diphosphate (ADP). The presence of endogenous ADP may be an important factor in clumping during standard concentrate preparation.A method of acidification of PRP using citric acid is described which allows preparation of an effective concentrate from fresh whole blood without subjecting the red cells to acid pH. Reconstitution of the acidified platelet poor plasma and its native red cells increases the citrate molarity by less than 6 per cent and results in minimal decrease in pH of the whole blood.

Resistance to bacteria in hemorrhagic shock. II. Effect of transient vascular collapse on sensitivity to endotoxin.

Summary and ConclusionRabbits transfused after one to 2 hours in hemorrhagic shock nearly always recover. During the immediate post transfusion recovery phase the sensitivity to a purified E. colt toxin is increased 100,000 times normal. Return to normal tolerance to the endotoxin requires about 48 hours.