Interlaboratory comparison of red-cell ATP, 2,3-diphosphoglycerate and haemolysis measurements.

BACKGROUND AND OBJECTIVES: Red blood cell (RBC) storage systems are licensed based on their ability to prevent haemolysis and maintain RBC 24-h in vivo recovery. Preclinical testing includes measurement of RBC ATP as a surrogate for recovery, 2,3-diphosphoglycerate (DPG) as a surrogate for oxygen affinity, and free haemoglobin, which is indicative of red cell lysis. The reproducibility of RBC ATP, DPG and haemolysis measurements between centres was investigated. MATERIALS AND METHODS: Five, 4-day-old leucoreduced AS-1 RBC units were pooled, aliquotted and shipped on ice to 14 laboratories in the USA and European Union (EU). Each laboratory was to sample the bag twice on day 7 and measure RBC ATP, DPG, haemoglobin and haemolysis levels in triplicate on each sample. The variability of results was assessed by using coefficients of variation (CV) and analysis of variance. RESULTS: Measurements were highly reproducible at the individual sites. Between sites, the CV was 16% for ATP, 35% for DPG, 2% for total haemoglobin and 54% for haemolysis. For ATP and total haemoglobin, 94 and 80% of the variance in measurements was contributed by differences between sites, and more than 80% of the variance for DPG and haemolysis measurements came from markedly discordant results from three sites and one site, respectively. In descending order, mathematical errors, unvalidated analytical methods, a lack of shared standards and fluid handling errors contributed to the variability in measurements from different sites. CONCLUSIONS: While the methods used by laboratories engaged in RBC storage system clinical trials demonstrated good precision, differences in results between laboratories may hinder comparative analysis. Efforts to improve performance should focus on developing robust methods, especially for measuring RBC ATP.

A prospective, randomized study of the use of platelet concentrates irradiated with ultraviolet-B light in patients with hematologic malignancy.

BACKGROUND: Irradiation of platelet concentrates (PCs) with ultraviolet-B (UVB) light inactivates the contaminating white cells and might be an alternative to filtration for the prevention of alloimmunization to HLA antigens and subsequent refractoriness to further platelet transfusions in multiply transfused patients with bone marrow failure. STUDY DESIGN AND METHODS: Patients with hematologic malignancy, mainly acute myeloid leukemia, were prospectively assigned in a random manner to receive either UVB-irradiated or control, nonirradiated PCs. All patients were given red cells that were white cell reduced by filtration. Transfusion efficacy and alloimmunization were assessed by means of corrected count increments, requirement for red cells and PCs, and measurement of lymphocyte-reactive antibodies. RESULTS: UVB-irradiated PCs had a clinical efficacy similar to controls as judged by corrected count increments at 1 to 6 and 12 to 24 hours and by the median requirement for red cell and platelet transfusions. Alloimmunization determined by measurements of lymphocyte-reactive antibodies using both conventional and antiglobulin-augmented lymphocytotoxicity techniques was not abolished in recipients of UVB-irradiated PCs (4/30, 13%) but was less than that in controls (5/20, 25%; p = NS). The mean number of platelet transfusion episodes prior to the occurrence of alloimmunization was greater in the control group (27 vs. 10; p = 0.017). CONCLUSION: In this trial, UVB irradiation did not diminish the clinical efficacy of platelet transfusions. There was a small but nonsignificant reduction alloimmunization, but no difference in refractoriness of the two groups was observed. Larger prospective randomized studies are required to confirm these findings and to compare UVB irradiation with white cell reduction.

Ultraviolet-B irradiation of platelets: a preliminary trial of efficacy.

Prior studies established that ultraviolet-B light (UVB) irradiation of platelet concentrates (PCs) at appropriate doses can eliminate the mixed lymphocyte culture-stimulating and -responding capacity of lymphocytes in the PCs without adversely affecting in vitro platelet function. The in vivo recovery and survival and in vitro characteristics of UVB-irradiated platelets were investigated in paired studies. PCs were stored for 1 day and then exposed to UVB. Platelet recovery, survival, and function were comparable to those of nonirradiated platelets. Recovery and survival of platelets stored for 5 days before UVB exposure were decreased relative to controls, although they were considered clinically acceptable. Paired transfusion studies were also performed in seven thrombocytopenic patients by using platelets obtained by apheresis. Comparable posttransfusion platelet increments and bleeding time corrections were obtained with both irradiated and control (nonirradiated) platelets. It can be concluded that platelets survive and function relatively normally in vivo after UVB irradiation sufficient to abolish lymphocyte reactivity in mixed lymphocyte culture. Long-term studies of UVB-irradiated PCs are needed to assess their potential in reducing recipient alloimmunization.

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.