RESUMO
BACKGROUND: Cold (4°C)-stored platelets are currently under investigation for transfusion in bleeding patients. It is currently unknown how long cold-stored platelets can be stored for clinical applications. STUDY DESIGN AND METHODS: Twenty three subjects were recruited. Twenty-one subjects were available for in vivo assessment and received indium-111 radiolabeled, cold-stored platelets. We investigated 5- (n = 5), 10- (n = 6), 15- (n = 5), and 20-day-stored (n = 5) platelets and obtained samples for in vitro testing at baseline and after the designated storage time. Twenty three units were available for in vitro testing. Five- and 7-day (n = 5 each), room temperature (RT)-stored platelets served as the current clinical standard control. RESULTS: In vivo, we found a continuous decline in platelet recovery from 5 to 20 days. Platelet survival reached a low nadir after 10 days of storage. Ex vivo, we observed the maximum platelet αIIbß3 integrin response to collagen at 5 days of cold storage, and we saw a continuous decline thereafter. However, platelet integrin activation and mitochondrial membrane integrity were better preserved after 20 days at 4°C, compared to 5 days at RT. Platelet metabolic parameters suggest comparable results between 20-day cold-stored platelets and 5- or 7-day RT-stored platelets. CONCLUSION: In summary, we performed the first studies with extended, cold-stored, apheresis platelets in plasma for up to 20 days with a fresh comparator. Storing cold-stored platelets up to 20 days yields better results in vitro, but further studies in actively bleeding patients are needed to determine the best compromise between hemostatic efficacy and storage prolongation.
Assuntos
Plaquetas/fisiologia , Preservação de Sangue/métodos , Criopreservação , Humanos , Ativação Plaquetária/fisiologia , Transfusão de Plaquetas , Plaquetoferese/métodos , Fatores de TempoRESUMO
BACKGROUND AND OBJECTIVES: Pathogen reduction of donor platelets with amotosalen/UVA has been shown to effectively inactivate pathogens and also contaminating white blood cells (WBCs). We wanted to determine whether WBC inactivation could also decrease alloimmune refractoriness to donor platelets. MATERIALS AND METHODS: Platelets were prepared from a donor dog's whole blood, and the platelets were either transfused without modification [standard (STD) platelets] or treated with amotosalen/UVA under conditions modelling the amotosalen/UVA Blood System for human platelets (APR) using either 4 or 3 J/cm2 of UVA exposure. Platelets were transfused weekly from a single donor dog for 8 weeks or until the recipient dog became refractory to their donor's platelets. Antibody samples were drawn weekly and tested against the donor dog's platelets and WBCs (CD8 and B cells). RESULTS: Only 1/7 (14%) dogs that received STD platelets accepted 8 weeks of donor transfusions. Following APR 4 J/cm2 donor transfusions, 3/9 (33%) recipients accepted their donor's transfusions, but only one recipient remained antibody negative. Following APR 3 J/cm2 donor transfusions, the same dose as used for human platelet transfusions, 7/10 (70%) recipients accepted their donor's transfusions, but only two remained antibody negative. CONCLUSION: As a very high percentage of recipient dogs (70%) accepted APR 3 J/cm2 donor transfusions, these data suggest that preventing alloimmune platelet refractoriness may be another benefit of pathogen reduction using amotosalen/UVA.
Assuntos
Doadores de Sangue , Transfusão de Sangue , Furocumarinas/farmacologia , Raios Ultravioleta , Animais , Cães , Feminino , Furocumarinas/uso terapêutico , Masculino , Modelos Animais , Transfusão de PlaquetasRESUMO
BACKGROUND: Ordinarily, whole blood (WB) is separated into components before storage. We assessed the posttransfusion viability and function of platelets (PLTs) if they were stored within WB at 4°C. STUDY DESIGN AND METHODS: Whole blood was obtained from 30 normal subjects and stored at 4°C without agitation for 12 days and for 10, 15, or 22 days with agitation. After WB storage, a PLT concentrate was prepared, and a fresh PLT sample was obtained from each donor. The stored PLTs were labeled with 111 In and the fresh with 51 Cr, and both were simultaneously transfused into their donor. Blood samples were obtained after transfusion to determine PLT recoveries and survivals. PLT samples from WB before and after storage were also assayed for PLT function and biochemistry. RESULTS: After storage for 12 days without WB rotation, poststorage PLT counts averaged only 49 ± 12% of baseline values. After storage for 10, 15, or 22 days with end-over-end WB rotation, PLT counts averaged 76 ± 14% of baseline values. Fifteen-day poststorage radiolabeled PLT recoveries averaged 27 ± 11% (49 ± 16% of fresh), and survivals averaged 1.2 ± 0.4 days (16 ± 6% of fresh). in vitro assays demonstrated marked PLT activation after any storage time, and although PLT function decreased over time, stored PLTs were still considered acceptable. CONCLUSION: These data suggest that, during rotated WB storage at 4°C for up to 15 days, PLT yields, poststorage PLT recoveries and survivals, and PLT function should be sufficient to support the short-term hemostatic needs of traumatized patients.
Assuntos
Plaquetas/citologia , Plaquetas/fisiologia , Preservação de Sangue/métodos , Hemostasia/fisiologia , Transfusão de Plaquetas/métodos , Plaquetoferese , Refrigeração , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Sobrevivência Celular , Temperatura Baixa , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Plaquetoferese/métodos , Refrigeração/métodos , Adulto JovemRESUMO
BACKGROUND: The current 5-day storage time of room temperature (22°C)-stored platelets (RSPs) severely limits platelet (PLT) availability. Extended cold (4°C)-stored PLTs (CSPs) are currently being investigated for actively bleeding patients. However, we currently do not know how to best store PLTs in the cold for extended periods of time. In this study, we investigate how storage in plasma and PLT additive solutions (PASs) affects PLT viability in vivo. STUDY DESIGN AND METHODS: Twenty normal subjects had a 2-unit hyperconcentrated apheresis PLT collection. One unit was stored at 4°C in plasma for 3 days ("control unit"), and the CSP "test" unit was stored for 10 or 15 days in plasma or 10 days in 35% plasma with either 65% Intersol or Isoplate. After storage, all units were radiolabeled and transfused into their donors. RESULTS: For 10-day storage, both the plasma and the Intersol units had significantly better PLT recoveries than the Isoplate units (24% ± 8% vs. 11% ± 3% [55% ± 11% vs. 21% ± 8% as percentage of control data], p = 0.002; and 18% ± 4% vs. 11% ± 3% [43% ± 6% vs. 21% ± 8% as percentage of control data], p = 0.004, respectively). There was a trend for lower PLT recoveries with Intersol compared to plasma (p = 0.056). PLT survivals and most in vitro measurements did not differ significantly among the units. CONCLUSIONS: While the in vitro variables suggest largely comparable results between plasma and PASs, in vivo recoveries were higher with plasma compared with both Intersol and Isoplate (p = 0.057 and p = 0.002, respectively). Whether this difference leads to clinically relevant differences in hemostatic efficacy remains to be determined.
Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Temperatura Baixa , Plaquetoferese/métodos , Transplante Autólogo/métodos , Sobrevivência Celular , Humanos , Transfusão de Plaquetas/métodos , Soluções , Fatores de TempoRESUMO
Human lymphocyte antigen alloimmunization to filter leukoreduced (F-LR) platelets occurs in about 18% of immunosuppressed thrombocytopenic hematology/oncology patients and represents a significant challenge for effective chemotherapy. In a dog platelet transfusion model, we have evaluated other methods of preventing alloimmune platelet refractoriness and demonstrated that successful methods in our dog model are transferable to man. In the present study, donor/recipient pairs were dog lymphocyte antigen DR-B incompatible (88% of the pairs), and recipient dogs received up to 8 weekly treated transfusions from a single donor (a highly immunogenic stimulus), or until platelet refractoriness. Continued acceptance of F-LR platelets occurred in 6 of 13 recipients (46%), but neither γ-irradiation (γ-I; 0 of 5) nor Mirasol pathogen reduction (MPR; 1 of 7) treatment of donor platelets prevented alloimmune platelet refractoriness. Combining γ-I with F-LR was associated with only 2 of 10 (20%) recipients accepting the transfused platelets. Surprisingly, F-LR platelets that then underwent MPR were accepted by 21 of 22 (95%) recipients (P < .001 vs F-LR + γ-I recipients). Furthermore, 7 of 21 (33%) of these accepting recipients demonstrated specific tolerance to 8 more weekly donor transfusions that had not been treated. In addition, platelet concentrates prepared from F-LR + MPR whole blood were also nonimmunogenic; that is, 10 of 10 (100%) recipients accepted donor platelets. Overall, 31 of 32 (97%) recipients accepted F-LR + MPR platelets; none developed antibodies to donor lymphocytes. These data are the highest rate of acceptance for platelet transfusions reported in either animals or man. This approach to platelet transfusion may be particularly important when supporting patients with intact immune systems, such as in myelodysplastic syndromes.
Assuntos
Plaquetas/imunologia , Transfusão de Sangue , Filtração , Imunização , Isoantígenos/imunologia , Leucócitos/citologia , Viabilidade Microbiana , Animais , Anticorpos/metabolismo , Cães , Tolerância Imunológica , Modelos Animais , Plasma Rico em Plaquetas/metabolismo , Análise de SobrevidaRESUMO
BACKGROUND: Alloimmune platelet (PLT) refractoriness remains a significant problem for chronically transfused patients with thrombocytopenia. STUDY DESIGN AND METHODS: In a dog PLT transfusion model, we evaluated ultraviolet B irradiation (UV-B) of donor PLTs-either alone or in combination with centrifuge leukoreduction (C-LR) or filtration leukoreduction (F-LR)-to prevent refractoriness to donor PLTs and to induce tolerance to standard (STD) PLTs from the same donor or to tertiary donors. RESULTS: Recipient acceptance rates for C-LR donor PLT transfusions were 14%, F-LR were 33%, and UV-B irradiated were 45% with no significant differences among the treatments given to the donor's PLTs. Adding UV-B irradiation to C-LR or F-LR PLTs increased acceptance rates to 50 and 68% (p = 0.02 and p = 0.05), respectively, comparing single treatments to the combined treatments. After a recipient had accepted any type of UV-B-treated donor PLTs, specific tolerance to subsequent transfusions of the same donor's STD PLTs averaged 65%. Nonspecific tolerance to third-party donor's STD PLTs averaged 36% if they had accepted their initial donor's treated PLTs but was only 4% (p < 0.001) if they had rejected these PLTs. CONCLUSION: Combining UV-B irradiation with a method of leukoreduction produces additive effects on prevention of alloimmune PLT refractoriness.
Assuntos
Tolerância Imunológica , Isoanticorpos/imunologia , Transfusão de Plaquetas/métodos , Raios Ultravioleta , Animais , Plaquetas/imunologia , Cães , Tolerância Imunológica/efeitos da radiação , Procedimentos de Redução de Leucócitos , Modelos AnimaisRESUMO
A literature review was conducted to assess the efficacy and safety of dimethyl sulfoxide (DMSO) cryopreserved platelets for potential military use. In vivo DMSO cryopreserved platelet studies published between 1972 and June of 2013 were reviewed. Assessed were the methods of cryopreservation, posttransfusion platelet responses, prevention or control of bleeding, and adverse events. Using the Department of Defense's preferred 6% DMSO cryopreservation method with centrifugation to remove the DMSO plasma before freezing at -65°C and no postthaw wash, mean radiolabeled platelet recoveries in 32 normal subjects were 33% ± 10% (52% ± 12% of the same subject's fresh platelet recoveries), and survivals were 7.5 ± 1.2 days (89% ± 15% of fresh platelet survivals). Using a variety of methods to freeze autologous platelets from 178 normal subjects, mean radiolabeled platelet recoveries were consistently 39% ± 9%, and survivals, 7.4 ± 1.4 days. More than 3000 cryopreserved platelet transfusions were given to 1334 patients. There were 19 hematology/oncology patient studies, and, in 9, mean 1-hour corrected count increments were 11 100 ± 3600 (range, 5700-15 800) after cryopreserved autologous platelet transfusions. In 5 studies, bleeding times improved after transfusion; in 3, there was either no improvement or a variable response. In 4 studies, there was immediate cessation of bleeding after transfusion; in 3 studies, patients being supported only with cryopreserved platelets had no bleeding. In 1 cardiopulmonary bypass study, cryopreserved platelets resulted in significantly less bleeding vs standard platelets. In 3 trauma studies, cryopreserved platelets were hemostatically effective. No significant adverse events were reported in any study. In summary, cryopreserved platelets have platelet recoveries that are about half of fresh platelets, but survivals are only minimally reduced. The platelets appear hemostatically effective and have no significant adverse events.
Assuntos
Plaquetas/citologia , Criopreservação/métodos , Dimetil Sulfóxido/química , Preservação de Sangue/métodos , Sobrevivência Celular , Raios gama , Hemostasia , Humanos , Transfusão de Plaquetas/métodos , TemperaturaRESUMO
BACKGROUND: Platelet (PLT) concentrates (PCs) prepared from whole blood in the United States are made using the PLT-rich plasma method. The PCs must be made within 8 hours of blood collection and stored for only 5 days. In Europe and Canada, PCs are made using the buffy coat (BC) method from whole blood held overnight at 22 °C and storage times may be up to 7 days. Our studies were designed to determine how long BC PLTs can be stored in plasma or Plasmalyte while meeting the FDA's poststorage viability criteria. STUDY DESIGN AND METHODS: Normal subjects donated whole blood that was stored at 22 °C for 22 ± 2 hours before preparation of BC PLTs. PLTs were stored for 5 to 8 days in either plasma or Plasmalyte concentrations of 65 or 80%. Radiolabeled autologous stored versus fresh PLT recoveries and survivals were assessed as well as poststorage in vitro assays. RESULTS: BC PLTs stored in either plasma or 65% Plasmalyte met FDA poststorage PLT recovery criteria for 7 days but survivals for only 6 days, while storage in 80% Plasmalyte gave very poor results. Both stored PLT recoveries and survivals correlated with the same donor's fresh results, but the correlation was much stronger between recoveries than survivals. In vitro measures of extent of shape change, morphology score, and pH best predicted poststorage PLT recoveries, while annexin V binding best predicted PLT survivals. CONCLUSION: BC PLTs stored in either plasma or 65% Plasmalyte meet FDA's poststorage viability criteria for 6 days.
Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Sobrevivência Celular/fisiologia , Humanos , Plasma Rico em Plaquetas/citologia , Fatores de TempoRESUMO
To evaluate the poststorage viability of apheresis platelets stored for up to 18 days in 80% platelet additive solution (PAS)/20% plasma, 117 healthy subjects donated platelets using the Haemonetics MCS+, COBE Spectra (Spectra), or Trima Accel (Trima) systems. Control platelets from the same subjects were compared with their stored test PAS platelets by radiolabeling their stored and control platelets with either (51)chromium or (111)indium. Trima platelets met Food and Drug Administration poststorage platelet viability criteria for only 7 days vs almost 13 days for Haemonetics platelets; ie, platelet recoveries after these storage times averaged 44 ± 3% vs 49 ± 3% and survivals were 5.4 ± 0.3 vs 4.6 ± 0.3 days, respectively. The differences in storage duration are likely related to both the collection system and the storage bag. The Spectra and Trima platelets were hyperconcentrated during collection, and PAS was added, whereas the Haemonetics platelets were elutriated with PAS, which may have resulted in less collection injury. When Spectra and Trima platelets were stored in Haemonetics' bags, poststorage viability was significantly improved. Platelet viability is better maintained in vitro than in vivo, allowing substantial increases in platelet storage times. However, implementation will require resolution of potential bacterial overgrowth during storage.
Assuntos
Plaquetas , Preservação de Sangue , Plaquetoferese , Soluções , Plaquetas/fisiologia , Preservação de Sangue/métodos , Dióxido de Carbono/metabolismo , Sobrevivência Celular , Glucose/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Consumo de Oxigênio , Plasma/química , Controle de Qualidade , Soluções/química , Fatores de TempoRESUMO
BACKGROUND: The Food and Drug Administration (FDA) requires that red blood cells must be refrigerated within 8 hours of whole blood collection. Longer storage of whole blood at 22°C before component preparation would have many advantages. STUDY DESIGN AND METHODS: Two methods of holding whole blood for 20 to 24 hours at room temperature were evaluated, refrigerated plates or a 23°C incubator. After extended whole blood storage, platelet (PLT) concentrates were prepared from PLT-rich plasma on Day 1 postdonation, and the PLTs were stored for 6 more days. On Day 7 of PLT storage, blood was drawn from each subject to prepare fresh PLTs. The stored and fresh PLTs were radiolabeled and transfused into their donor. RESULTS: Eleven subjects' whole blood was stored using refrigerated butanediol plates (Compocool, Fresenius), and 10 using an incubator. Poststorage PLT recoveries averaged 47 ± 13% versus 53 ± 11% and survivals averaged 4.6 ± 1.7 days versus 4.7 ± 0.9 days for Compocool versus incubator storage, respectively (p = NS). With all results, poststorage PLT recoveries averaged 75 ± 10% of fresh and survivals 57 ± 13% of fresh; PLT recoveries met FDA guidelines for poststorage PLT viability but not survivals. CONCLUSION: Seven-day poststorage PLT viability is comparable when whole blood is stored for 22 ± 2 hours at 22°C using either refrigerated plates or an incubator to maintain temperature before preparing PLT concentrates.
Assuntos
Preservação de Sangue/métodos , Temperatura Baixa , Plasma Rico em Plaquetas/fisiologia , Plaquetoferese/métodos , Plaquetas/citologia , Plaquetas/fisiologia , Coleta de Amostras Sanguíneas/métodos , Temperatura Corporal/fisiologia , Sobrevivência Celular , Humanos , Contagem de Plaquetas , Plasma Rico em Plaquetas/citologia , Refrigeração , Fatores de TempoRESUMO
BACKGROUND: A recent review concluded that there was inadequate evidence to show a difference between buffy coat (BC) and platelet (PLT)-rich plasma (PRP) PLT concentrates prepared from whole blood. We hypothesized that 7-day-stored BC-PLTs would have superior autologous recoveries and survivals compared to PRP-PLTs and that both would meet the Food and Drug Administration (FDA) criteria for poststorage viability. STUDY DESIGN AND METHODS: This was a randomized, crossover study design in healthy subjects who provided informed consent. Each participant donated a unit of whole blood on two occasions. In random order, either BC-PLTs or PC-PLTs were prepared after a 20 ± 2 °C overnight hold of the whole blood. PLTs were stored under standard conditions. On Day 7, fresh PLTs were prepared from 43 mL of autologous whole blood. The fresh PLTs paired with either BC-PLTs or PRP-PLTs were alternately labeled with (111) In or (51) Cr and simultaneously reinfused to determine recoveries and survivals. In vitro assays were performed on Days 1 and 7. RESULTS: Fourteen subjects completed the study at two sites. No differences in poststorage PLT viabilities were observed between BC-PLTs and PRP-PLTs; recovery differences averaged 3.7 ± 2.4% (± SE, p = 0.15) and survival differences averaged 0.48 ± 0.56 days (p = 0.41). Neither type of PLTs met the current FDA criteria for either poststorage PLT recoveries or survivals. CONCLUSION: We were unable to demonstrate that single-unit BC-PLTs stored for 7 days have superior poststorage viability compared to PRP-PLTs. Failure to meet the minimum FDA criteria for poststorage PLT viability raises questions regarding the acceptance thresholds of these metrics.
Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Plasma Rico em Plaquetas/citologia , Preservação de Sangue/efeitos adversos , HumanosRESUMO
BACKGROUND: Using bacterial detection or pathogen reduction, extended platelet (PLT) storage may be licensed if PLT viability is maintained. The Food and Drug Administration (FDA)'s poststorage PLT acceptance guidelines are that autologous stored PLT recoveries and survivals should be 66 and 58% or greater, respectively, of each donor's fresh PLT data. STUDY DESIGN AND METHODS: Nonleukoreduced PLT concentrates were prepared from whole blood donations. Autologous PLT concentrates from 62 subjects were stored in 100% plasma (n=44) or 20% plasma/80% Plasmalyte (n=18), an acetate-based, non-glucose-containing crystalloid solution previously used for PLT storage. Fresh PLTs were obtained on the day the donor's stored PLTs were to be transfused. The fresh and stored PLTs were alternately radiolabeled with either (51) chromium or (111) indium, and in vitro measurements were performed on the stored PLTs. RESULTS: The FDA's PLT recovery criteria were met for 7 days of plasma storage, but PLT survivals maintained viability for only 6 days. Plasmalyte-stored PLTs did not meet either acceptance criteria after 6 days of storage. After 7 days of storage, PLT recoveries averaged 43±4 and 30±4% and survivals 4.1±0.4 and 2.0±0.2 days for plasma- and Plasmalyte-stored PLTs, respectively (p=0.03 for recoveries and p<0.001 for survivals). Poststorage PLT recoveries correlated with the commonly used in vitro PLT quality measurements of hypotonic shock response and annexin V binding, while survivals correlated with extent of shape change, morphology score, and pH. CONCLUSION: There is a progressive decrease in recoveries and survivals of plasma-stored PLTs over time. PLT viability is better maintained in plasma than Plasmalyte.
Assuntos
Plaquetas/citologia , Plaquetas/metabolismo , Preservação de Sangue/métodos , Plasma Rico em Plaquetas , Sobrevivência Celular/fisiologia , Humanos , Fatores de TempoRESUMO
BACKGROUND: Three of four prior studies suggested that warming platelets (PLTs) to 37 degrees C before transfusion into patients with thrombocytopenia gave improved corrected PLT count increments. STUDY DESIGN AND METHODS: Eighteen normal subjects had apheresis PLTs collected that were stored at 22 degrees C for 5 days in two storage bags. One bag of PLTs was warmed to 35 degrees C before infusion, and one remained at 22 degrees C. Three different methods of warming the donor's autologous PLTs before reinfusion were evaluated: warming PLTs to 35 degrees C for 10 or 60 minutes followed by radiolabeling or radiolabeling the PLTs followed by warming to 35 degrees C for 60 minutes. In the first two methods, the warmed PLTs would have returned to 22 degrees C before infusion, and in the third, the PLTs would still be warm when injected. The paired test and control PLTs were radiolabeled with either (111)In or (51)Cr to determine posttransfusion PLT recoveries and survivals. PLT morphology score, pH, hypotonic shock response, extent of shape change, and annexin V binding were determined just before transfusion. RESULTS: There were no differences in posttransfusion autologous radiolabeled PLT recoveries and survivals or in the in vitro measurements for the PLTs maintained at 22 degrees C versus those warmed to 35 degrees C using any of the three methods of PLT warming before infusion. CONCLUSION: Based on these 5-day-stored autologous radiolabeled PLT recovery and survival measurements, there is no evidence that warming PLTs to 35 degrees C before infusion improves postinfusion PLT viability.
Assuntos
Plaquetas/citologia , Preservação de Sangue/efeitos adversos , Sobrevivência Celular/fisiologia , Transfusão de Plaquetas , Humanos , TemperaturaRESUMO
BACKGROUND: Methods of bacterial detection and pathogen inactivation of platelets (PLTs) may allow extended storage of PLTs as long as PLT quality is maintained. STUDY DESIGN AND METHODS: Twenty normal volunteers had their PLTs collected with an apheresis machine (Haemonetics Corp.). A variety of in vitro PLT function and metabolic assays were performed both on Day 0 and after 8 days of storage. On Day 8, a small blood sample was drawn from each donor to obtain fresh PLTs. The fresh and stored autologous PLTs were labeled with either (51)Cr or (111)In, and the radiolabeled PLTs were transfused. Posttransfusion serial blood samples were drawn to determine the relative posttransfusion recoveries and survivals of the fresh versus the stored PLTs. RESULTS: Although the in vitro assays showed some differences between the two trial sites, the results were generally within the ranges expected for fresh and stored PLTs. Overall, PLT recoveries averaged 66 +/- 16 percent versus 53 +/- 20 percent and survivals averaged 8.5 +/- 1.6 days versus 5.6 +/- 1.6 days, respectively, for fresh compared to 8-day-stored PLTs. There were no significant differences in the in vivo PLT data between the trial sites or based on the radiolabel used for the measurements. CONCLUSION: After 8 days of storage, the in vivo posttransfusion recovery and survival of autologous Haemonetics apheresis PLTs meet the proposed standards for poststorage PLT quality.