Effect of Platelet Concentrates (PCs) Leucodepletion on the Activation and Efficacy of Platelet Transfusion (original) (raw)
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Pan African Medical Journal
Introduction: For many years, platelet concentrates have been used for the prevention as well as treatment of bleeding disorders, especially in those patients with haematological problems involving platelet disorders as well as refractoriness, In addition, platelet concentrates (PCs) have been widely used to support patients undergoing bone marrow transplantation or who are receiving myelotoxic treatments. The aim of this study was to determine the quality of platelet concentrates by assessing platelet counts, volume, pH changes, swirling, residue of the red blood cells and white blood cell counts. Assess the in vivo viability of a transfused platelet product using the corrected count increment (CCI) and the percentage platelets response (PPR). This descriptive analysis study was done in Kenyatta National Hospital Blood Transfusion Unit between July 2016 and December 2016. Methods: The in vitro Platelets concentrates quality was accurately determined and assessed using certain parameters. Platelet concentrates in ethylene diamine tetra acetic acid (EDTA) was used for analysis using Cell-Dyn 3700 analyser. The volume of PCs used was an average of 2mls of PCs, the pH was measured using digitalised Hanna edge pH kit. Agitation was done using Helmer agitator and centrifugation was done using Roto silenta 630 RS centrifuge. The in vivo viability of a transfused product was determined using corrected count increment (CCI) and percentage recovery (PR) between 1 and 20-hour after transfusion. Pre and post-transfused whole blood in EDTA collected from the recipients was analysed to access the functional platelets in the circulation. Data analysis was done using SPSS. Results: A total of 384 platelet concentrates were analysed and used in transfusion. The majority 96, (40%) were O Rhesus D+ and the least being AB Rhesus D-at (1%).Centrifugation, separation and agitation was done according to standard procedure (n=384). Only (246 (65%) of the concentrates were found fit for use out of a total of (n=384) leaving 138 (35%) which did not meet the KNH/KNBTS criteria. The minimum specifications for platelet count are 5.5 x (10 9). The duration of 3 days of storage on average, the WBC count (10 9) was Mean ± SD 4.50 ± 3.50. Using the Hanna edge pH kit the pH Mean was ±SD 7.18 ± 8.82 and the used Volume (Mls) was at 55 ± 15. The concentrate was issued within 3 days of processing. After transfusion, the percentage platelet response (PPR) was 72% in male recipients at 1-hour and 30% at 20-hours while 69% in female recipients at 1-hour and 25% at 20-hours. The invivo viability of platelet product had a corrected count increment (CCI) of 75% ≥ 7500 at 1-hour and CCI of18% ≥ 30% at 20-hours in male recipients. In the same study, the female recipients had a CCI of 80% ≥ 7500 at 1-hour and a CCI of 25% ≥ 30% at 20-hours. Conclusion: The findings on platelets concentrates quality 65% met platelets transfusion criteria while 35% did not. On preparation of platelets concentrates there was high counts of white blood cells 4.5±3.5×10 9 than recommended counts by Kenya National Blood Transfusion Services < 0.83×10 9. Both percentage platelet response (PPR) and corrected count increment (CCI) were very low at 20 hours compared to British committee for standards haematology criteria for successful increment of platelet products (PPR ≥ 30% and CCI ≥ 7500). Apheresis platelets transfusion can be introduced at KNH and use of leukoreduction performed on the platelet concentrates which are prepared within the Hospital. With such rate of refractoriness, additional tests to confirm the real cause of unviability of platelets in the patients need to be performed. Recipients should be done evaluation of the pattern of refractoriness followed by HLA compatibility testing. In addition, if there is a high, compatible cross-matched, selected apheresis platelet concentrate pint should be transfused. This unviability was due to recipients with either immune-mediated refractoriness or non-immune mediated refractoriness.
Transfusion, 2008
BACKGROUND: One of the variables to determine the quality of platelets (PLTs) in vitro is measurement of CD62P expression. Different protocols are in use, however, making comparison of results virtually impossible. It was our aim to develop a uniform CD62P protocol that would yield comparable results in various laboratories. STUDY DESIGN AND METHODS: The effects of fixation, source and dilution of CD62P antibody, source of immunoglobulin G (IgG) isotypic antibody, and analysis of results were investigated. Once the optimal variables were defined, comparative studies were performed at five participating centers. In the final comparative study, eight split PLT concentrates were shipped to the centers, where samples were stained and fixed according to the uniform protocol. Analyses were performed using commercially available flow cytometers (BD Biosciences and Beckman Coulter). RESULTS: Uniformity between centers could be achieved by using a single clone for CD62P and IgG monoclonal antibody. A protocol was selected using fixation with 0.5 percent methanol-free formaldehyde. To increase conformity between flow cytometers, in the analysis of electronic data the thresholds of the isotypic control were set at 0.5 percent for the BD Biosciences and 2 percent for the Beckman Coulter flow cytometers. In the final comparative study, the 95 percent confidence intervals (CIs) for CD62P ranged between 8 and 21 percent in fresh and 20 to 40 percent in 8-day-old PLT concentrates. CONCLUSION: A uniform CD62P staining protocol and subsequent analysis can be used at multiple centers using different flow cytometers, yielding comparable results with acceptable 95 percent CIs.
Automated CD61 immunoplatelet analysis of thrombocytopenic samples
British Journal of Haematology, 2001
Revision of the current decision point for prophylactic platelet transfusion in thrombocytopenic patients requires the availability of a method that is able to provide accurate platelet counts to as low as 1 Â 10 9 /l. This study is the first to evaluate the immunoplatelet method (CD61-Imm) of the haematological analyser Cell-Dyn 4000 in direct comparison with the flow cytometric procedure. Additionally CD61-Imm results were compared with CD4000 optical (Plto) counts in the ranges 20±547 Â
Journal of thee Medical Sciences (Berkala Ilmu Kedokteran), 2017
Thrombocyte concentrate (TC) transfusion is an important supportive therapy in patients with thrombocytopenia. The risks in platelet transfusions may be related to the content of TC including the contaminant leukocytes. The aim of this study was to assess the risk of increased level of P-Selectin (CD62P) expression of non-leukodepleted TC transfusions. This was a quasi-experimental study. Subjects were children patients aged 1-18 years who received a non-leukodepleted or a leukodepleted TC transfusions. Comparison of the proportion of increased expression of CD62P in both groups expressed as relative risk. The subjects consisted of 51 patients who received non-leukodepleted and 52 patients who received leukodepleted TC transfusions. The risk of increased expression of CD62P in patients receiving non-leukodepleted TC transfusions were 2.38 (95%CI:1.60-3.53) times higher than those who received leukodepleted TC. Non-leukodepleted have significant higher risks of increased CD62P expression than leukodepleted TC transfusions.
Transfusion, 2010
Clinical effect of platelet (PLT) transfusion is monitored by measures of PLT viability (PLT recovery and survival) and functionality. In this study we evaluate and compare transfusion effect measures in patients with chemotherapy-induced thrombocytopenia due to treatment of acute leukemia. Forty transfusions (28 conventional gamma-irradiated and 12 pathogen-inactivated photochemical-treated PLT concentrates [PCs]) were investigated. PC quality was analyzed immediately before transfusion. Samples were collected from thrombocytopenic patients at 1 and 24 hours for PLT increments and thromboelastography (TEG) with assessments of bleeding score and intertransfusion interval (ITI). Data were analyzed by Spearman&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s correlation. Patient and PC variables influencing the effect of transfusion were analyzed by use of a mixed-effects model. PLT dose, storage time, and pathogen inactivation correlated with PLT recovery but not with PLT survival (including ITI), TEG, or clinical bleeding. Fever was negatively correlated with PLT survival but did not affect PLT recovery. After 1 and 24 hours, strong correlations were observed within measures of PLT viability and between PLT increment and the TEG value maximal amplitude (MA). Negative correlation was observed between late MA increment and clinical bleeding status after transfusion (r = -0.494, p = 0.008). PLT count increments did not correlate to clinical bleeding status. PLT dose and quality of PCs are important for optimal immediate transfusion response, whereas duration of transfusion effect is influenced mainly by patient variables. The TEG value MA correlates with PLT count increments and bleeding, thus reflecting both PLT viability and functionality.
Blood transfusion = Trasfusione del sangue, 2014
Accurate platelet counts (PC) are necessary in order to follow recommendations for prophylactic platelet transfusion. We carried out a study comparing the standard way of counting platelets using a routine analyser and compared it with PC determined by flow cytometry (FC) and haemostatic data obtained with thromboelastography (TEG). The study was carried out on 24 patients with haematological malignancies, all given one adult dose of platelets. The PC was determined before and after transfusion using an automated blood cell counter and FC. Citrated, "native" whole blood TEG was carried out before and after platelet transfusion to assess global haemostasis. No bleeding was observed in any of the subjects. Thirty-one assessments were performed in the 24 patients. The mean pre-transfusion PC were 9.8 and 13×10(9)/L with the automated counter and FC, respectively with a difference of 3.7 (p=0.0011). Excellent correlation was observed between the two counts (r=0.89; p<0.0001...
In vitro assessment of platelet storage lesion in leucoreduced random donor platelet concentrates
PubMed, 2010
Background: Currently platelet concentrates (PC) are collected using different synthetic materials and different centrifugation/leucocyte-removal processes. Upon exposure to artificial surfaces and high centrifugation forces, blood cells can undergo various levels of stress-induced, cellular activation/fragmentation and release reactions which may not only influence the extent of the platelet storage lesion but may also contribute to poor clinical effectiveness of the PC and transfusion reactions. Materials and methods: An array of assays, used for quality control of PC, was performed in two different groups of PC prepared from random donor plasma on days 1, 3 and 5 of storage. The group 1 PC were not leucoreduced while the group 2 PC underwent prestorage leucoreduction using a PL50E filter. As current recommendations for the evaluation of PC include the measurement of platelet activation, in this study CD62P on platelet membrane was measured. Furthermore, in vitro studies indicate that sHLA antigens may modulate immune competent cell function so, the presence of sHLA-1 in blood components is considered a marker of immunological reactivity and this, too, was measured. Results: The levels of CD62P and sHLA-1 were significantly lower in leucoreduced PC than in non-leucoreduced ones. However, the overall rate of increase of sHLA-1 during storage was faster in the leucoreduced group of PC. No significant differences were detected regarding other assays of quality. Conclusion: Based on our findings, leucoreduced PC differ from non-leucoreduced ones in terms of some specific markers such as CD62P as a marker of platelet activation and sHLA-1 as a marker of immunological reactivity. Pre-storage leucofiltration, followed by storage in currently used plastic bags is a safe procedure for PC for up to 5 days. The available leucoreduction technologies are not, however, sufficiently robust to completely abrogate transfusions reactions, and improvements are required to reach the goal of optimised yield and minimal transfusion reactions with platelet therapy.
Blood Reviews, 2011
Platelet transfusions are mainly used for patients with thrombocytopenia due to bone marrow failure, especially cancer patients developing severe chemotherapy-induced thrombocytopenia (e.g. patients with acute leukemia or other hematologic malignancies). A prophylactic transfusion strategy is now generally accepted in developed countries. Some clinical data, however, support the use of a therapeutic transfusion strategy at least for certain subsets of these patients. Several methodological approaches can then be used to evaluate the outcome of platelet transfusions, including peripheral blood platelet increments and bleeding assessments. Several factors will influence the efficiency of platelet transfusions; fever and ongoing hemorrhage are among the most important patient-dependent factors, but the number and quality of the transfused platelets are also important. The quality of transfused platelets can be evaluated by analyzing platelet activation, metabolism or senescence/apoptosis. Only evaluation of metabolism is included in international guidelines, but high-throughput methods for evaluation of activation and senescence/apoptosis are available and should be incorporated into routine clinical practice if future studies demonstrate that they reflect clinically relevant platelet characteristics. Finally, platelet transfusions have additional biological effects that may cause immunomodulation or altered angioregulation; at present it is not known whether these effects will influence the long-time prognosis of cancer patients. Thus, several questions with regard to the optimal use of platelet transfusions in cancer patients still need to be answered.
Evaluation of platelet turnover by flow cytometry
Platelets, 2006
The number of circulating newly produced platelets depends on the thrombopoietic capacity of bone marrow as well as platelet removal from the bloodstream. Flow cytometric analysis with thiazole orange (TO), a fluorescent dye that crosses platelet membranes and binds intracellular RNA, has been used to measure circulating reticulated platelets (RPs) with high RNA content as an index of platelet turnover. We first assessed the specificity of TO flow cytometry and then applied this method in the diagnosis of thrombocytopenia caused by impaired platelet production or increased destruction. We also explored the utility of TO flow cytometry to predict thrombocytopoiesis after chemotherapy-induced bone marrow aplasia. Venous blood, anticoagulated with K 2 EDTA, was incubated with 0.6 mg/ml TO plus an anti-GPIIIa monoclonal antibody. The mean percentage of RPs in control subjects (n ¼ 23) was 6.13 AE 3.09%. RPs were 10.41 AE 9.02% in patients (n ¼ 10) with hematological malignancies during aplasia induced by chemotherapy and a significant increase in RPs (35.45 AE 6.11%) was seen in the recovery phase. In 10 patients with idiopathic thrombocytopenic purpura, the percentage of TO positive platelets was 67.81 AE 18.79 (P < 0.001 vs. controls). In patients with thrombocytopenia associated with hepatic cirrhosis (n ¼ 21; 21.04 AE 16.21%, P < 0.001 vs. controls) or systemic lupus erythematosus (n ¼ 6, 29.08 AE 15.57%; P < 0.001 vs. controls) increases in TO-stained platelets were also observed. Measurement of TO positive platelets may be a reliable tool for the laboratory identification of platelet disorders, with a higher sensitivity than measurement of platelet volume. Measurement of RPs may also prove useful to recognize the underlying pathogenetic mechanisms in thrombocytopenia.