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PubMed2005

The impact of two whole blood inline filters on markers of coagulation, complement and cell activation.

BACKGROUND AND OBJECTIVES: There exists a current lack of information about the impact of different inline filters, used for the leucoreduction of whole blood (WB), on the levels of clotting factors and markers of coagulation, complement and cell activation in plasma. Only a few small comparisons of different types of WB inline filters have been published to date. MATERIALS AND METHODS: This study compared two plasma types of 200 units each. Both study groups were derived from WB, inline-filtered and held for 2 h at 20 degrees between donation and filtration. Then, 200 units (Group A) were filtered using a positively charged polyester filter (Baxter RZ2000) and the other 200 units (Group B) were filtered using an uncharged polyester filter (Fresenius). After filtration, both groups were analysed for fibrinogen, factors V and VIII:C (FV and FVIII:C, respectively), immunoglobulin G (IgG), residual leucocytes and platelets, and markers of coagulation, complement and cell activation. Predonation plasma samples from CPDA1-anticoagulated blood were obtained from 100 different individuals and served as controls. RESULTS: WB inline filtration did not influence fibrinogen, FV, FVIII:C or IgG levels. Neither filter induced thrombin or fibrin formation. The charged filter caused substantial complement activation and neutrophil elastase and platelet factor 4 release. In contrast, the plasma filtered through the uncharged filter showed markedly lower levels of C3a-desArg, C5a, neutrophil elastase and platelet factor 4, and moderately reduced levels of prothrombin fragments 1+2 and D-dimer, compared with controls. CONCLUSIONS: Filter type has a significant impact on the quality of plasma derived from WB filtered through inline filtration systems. Some filters produce substantial coagulation and complement activation and cell release, while others appear to reduce the plasma levels of activation markers. The clinical significance of these findings remains to be determined.

Adolescent↗
PubMed2005

Gamma irradiation does not affect 7-day storage of platelet concentrates.

BACKGROUND AND OBJECTIVES: Extension of storage of leucocyte-reduced platelet concentrates (LR-PCs) to 7 days has recently been introduced, but the effect of gamma irradiation on in vitro parameters was unknown. MATERIALS AND METHODS: A paired experiment was carried out, with gamma irradiation on day 1 or day 5 with 25 Gy, and no irradiation served as a reference. RESULTS: Measurements during storage up to 8 days after blood collection showed no significant differences of irradiated LR-PCs vs. the control. CONCLUSIONS: Irradiation, either on day 1 or day 5, had no effect on platelet quality during a 7-day storage period.

Blood Platelets↗
PubMed2005

In-vitro evaluation of the PALL Leukotrap Affinity Prion Reduction Filter as a secondary device following primary leucoreduction.

BACKGROUND AND OBJECTIVES: A filter (PRF1) designed to remove abnormal prion proteins from red-cell units has been developed. The purpose of this study was to evaluate the quality of red cells produced using this device. MATERIALS AND METHODS: Leucocyte-depleted red-cell units (CPD, CPD-A1 and CPD/SAGM) processed according to standard UK practices were filtered using PRF1. Filtered and control units were stored and sampled on day 1, day 7 and on the date of expiry and were tested using standard measures of red-cell quality. RESULTS: Filtered units were found to have significantly higher percentage haemolysis levels, lower haemoglobin levels and a smaller volume compared with controls. All results, however, were well within the permitted 0.8% haemolysis level at the end of storage and all units met the UK guidelines for haemoglobin and volume. The other test parameters measured showed no significant differences between the test and control units. CONCLUSIONS: The PRF1 filter was found to be easy to use and resulted in red-cell units that met all relevant UK and European Guidelines.

Erythrocyte Transfusion↗
PubMed2007

Portable microscopic cell counter for the determination of residual leucocytes in blood components.

BACKGROUND AND OBJECTIVES: The accurate determination of residual white blood cell (WBC) in blood components is of considerable clinical importance, and a variety of methods have been devised for the counting of low levels of residual WBC. In this study, we evaluated the performance of microscopic cell counter with microchannel plastic chip (C-reader) with regard to its ability to quantify WBC in WBC-reduced red cell concentrates. MATERIALS AND METHODS: In order to quantify residual WBC with the microscopic cell counter, WBC-reduced red cell concentrate was stained using propidium iodide. Three studies were performed: linearity, precision and correlation compared to those of manual Nageotte chamber counting and automatic flow cytometric methods. RESULTS: Dilution experiments, conducted over a range of 0.7-712 WBC/microl, showed a linearity of r(2) > 0.999, with coefficient of variation values of < or = 15.6% and accuracy of 93.8% over all tested ranges. In comparison with the Nageotte chamber counting and flow cytometric methods, the correlation coefficients were r(2) > 0.995. The detection limit of this method was 0.24 WBC/microl. Total analysis time per sample was approximately 5 min. CONCLUSION: The microscopic cell counter for residual WBC counting was determined to be efficient at the level of currently defined standards, with acceptable precision and accuracy. This method may prove useful for the quality assurance and control of WBC-depleted blood products.

Blood Component Transfusion↗
PubMed2005

Partial depletion of macrophages by ED7 reduces renal injury in Adriamycin nephropathy.

BACKGROUND: Because macrophages are considered to be possible effectors of disease in Adriamycin (ADR) nephrosis, we hypothesized that depletion of macrophages might protect against the initiation of renal injury. In the present study, a monoclonal antibody (ED7) directed against CD11b/CD18 integrin, which is expressed by macrophages, was used to investigate the pathogenetic effects of macrophages in ADR nephropathy. METHODS: Male Wistar rats were treated with ED7 antibody, starting 1 day prior to ADR (7.5 mg/kg) treatment, or 7 days post-ADR when overt proteinuria was established. RESULTS: Circulating ED7-positive cells were reduced by approximately 30% in rats with ADR nephrosis by the ED7 antibody, while the number of macrophages in the renal cortex of ADR rats was reduced by nearly 50% with the ED7 treatment, whether administered before or after ADR. Creatinine clearance was significantly ameliorated by ED7 when commenced pre-ADR (P < 0.05), but not when commenced post-ADR (P = NS) in comparison to untreated ADR rats. However, proteinuria was not alleviated by either ED7 treatment. Morphometric analysis showed less glomerular sclerosis when ED7 was commenced pre-ADR compared with ADR alone (P < 0.01), but not when commenced post-ADR (P = NS). Tubular atrophy was reduced by ED7 when it was commenced pre-ADR (tubular cell height and tubular diameter: P < 0.01 and P < 0.001, respectively), as was interstitial expansion (P < 0.01) compared with ADR alone. Cortical macrophage infiltration was reduced by 50% compared with ADR alone by the ED7 commenced before or after ADR. The number of cortical CD4+ T cells fell with ED7 starting pre-ADR, but not with the ED7 treatment commencing after ADR. CONCLUSION: Partial macrophage depletion starting before but not after ADR protected both renal function and structure in this model of chronic proteinuric renal disease.

Animals↗
PubMed2006

Clinical efficacy and biocompatibility of three different leukocyte and fat removal filters during cardiac surgery.

Activated leukocytes and fat particles are associated with organ injury after a cardiac surgery. Filters are currently used to remove either leukocytes or fat particles. A novel approach with a filter that combines leukocyte and fat removal might be clinically useful. As it is not known which type of filter has a good and safe performance in both leukocyte and fat removal, we measured in this study the leukocyte and fat removal properties and the biocompatibility of three different filters. We used six Pall RS1 (Pall, Portsmouth, England) leukocyte removal filters, six Pall LipiGuard fat removal filters, and six Fresenius Biofil 02 (Fresenius, Emmer-Compascuum, The Netherlands) leukocyte removal filters and measured the passage times of 500 and 1000 mL of residual heart-lung machine blood. We determined the circulating leukocyte and platelet counts, and total hemoglobin, triglyceride, and free fatty acid concentration after the filters. In addition, we measured free hemoglobin, plasma elastase (Merck, Darmstadt, Germany), and complement C5-9 (Quidel, San Diego, CA, U.S.A.) to assess the biocompatibility of the filters. The circulating fat particles were calculated with an automated hematology analyzer. The passage time for the blood was shortest for the Biofil filter (P = 0.02, analysis of variance). The total leukocyte counts (P = 0.04) and fat particles (P = 0.02) were higher after the LipiGuard filter. This filter also had a higher increase in free hemoglobin concentration (P = 0.03). We conclude that the leukocyte removal filters were superior to the fat removal filter both in leukocyte and fat removal.

Aged↗
PubMed2005

High-level long-term white blood cell microchimerism after transfusion of leukoreduced blood components to patients resuscitated after severe traumatic injury.

BACKGROUND: Long-term white blood cell (WBC) microchimerism (MC), of at least 2 years, has been reported in trauma patients receiving fresh nonleukoreduced (non-LR) blood. It is unknown, however, whether this occurs with LR blood products that are nearly devoid of WBCs. Twenty-seven patients transfused with LR and non-LR blood products were studied after severe traumatic injury. A secondary aim was to explore donor-recipient mixed lymphocyte reactivity in vitro. STUDY DESIGN AND METHODS: To quantify MC, allele-specific real-time polymerase chain reaction assays were developed targeting HLA Class II sequence polymorphisms. Extensive validation showed that these assays reliably detect a single copy of target sequence in a complex allogeneic background without false positivity. RESULTS: At a median follow-up of 26 months (range, 24-39 months), long-term MC was observed in 3 of 20 patients (15%) who received non-LR blood products and 2 of 7 (29%) who received LR blood products. The maximum MC ranged from 0.40 to 4.90 percent of circulating WBCs and appeared, by Class II genotype analysis, to be attributable to a single donor. CONCLUSION: It is concluded that robust levels of long-term MC, apparently traceable to a single donor, occur at similar frequency despite leukoreduction of transfused blood products. Exploratory analysis of donor-recipient mixed lymphocyte reactivity suggests that long-term MC may require a state of bidirectional tolerance before transfusion.

Blood Component Transfusion↗
PubMed2005

In vitro and in vivo evaluation of leukoreduced platelets stored for 7 days in CLX containers.

BACKGROUND: Extension of platelet (PLT) storage and concomitant use of a bacterial detection system would provide logistical advantages by reducing outdating and improving patient care through promotion of the use of sensitive detection systems. This study evaluated the in vitro characteristics and in vivo viability of leukoreduced PLT units derived from PLT-rich plasma stored for 5 days (control) versus 7 days (test) in CLX plastic containers. STUDY DESIGN AND METHODS: Two whole-blood units were collected from each subject into a leuko-reduction filtration system (Leukotrap RC-PL system, Pall Medical) in a paired design, the second 2 days after the first. These were leukoreduced (Leukotrap PL) and stored for 7 and 5 days. Poststorage samples from test and control units were randomly labeled with (51)Cr or (111)In and simultaneously infused autologously to determine recovery and survival. RESULTS: Small but significant (p < 0.05, paired t test) differences between 5 and 7 days of storage were seen in in vitro variables such as extent of shape change, hypotonic shock response, morphology, and P-selectin expression. In vivo recovery declined on average 11 percent with the two additional days of storage from 54.4 +/- 13.6 to 48.7 +/- 15.0 percent (p < 0.002); survival decreased on average 19 percent from 6.7 +/- 1.0 to 5.4 +/- 1.7 days (p < 0.002). CONCLUSION: Storage for 7 days was associated with reduced recovery and survival and in vitro variables, suggestive of extension of the storage lesion. These differences, however, were small in magnitude and unlikely to have significant clinical effects. Current collection and storage systems provide PLTs that are as functional at 7 days as those licensed for 7-day storage two decades ago.

Blood Platelets↗
PubMed2005

Evaluation of BacT/ALERT plastic culture bottles for use in testing pooled whole blood-derived leukoreduced platelet-rich plasma platelets with a single contaminated unit.

BACKGROUND: In certain countries, whole blood-derived platelet (PLT)-rich plasma PLTs can only be pooled within 4 hours of transfusion. One prerequisite for prestorage pooling is the ability to detect low levels of bacteria from a single unit (approx. 10 colony-forming units [CFUs]/mL) once pooled (10/6 approx. 2 CFUs/mL). This study evaluated the BacT/ALERT (bioMérieux) for detection of bacteria in 1 unit of a 6-unit pool. STUDY DESIGN AND METHODS: Bacillus cereus, Clostridium perfringens, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Serratia marcescens, Streptococcus viridans, and Propionibacterium acnes were inoculated into single PLT units (target, 10 and 100 CFUs/mL; mean recovered, 5 and 92 CFUs/mL) and then pooled with 5 sterile units. Four milliliters was inoculated into both plastic aerobic and anaerobic bottles, and 0.5 mL was plated (10 sets). RESULTS: All cases were detected when the single unit had at least 6 CFUs per mL. With B. cereus (< or =2 CFUs/mL), all bottles were reactive. With K. pneumoniae and S. viridans (< or =3 CFUs/mL), all samples were detected with a two-bottle set, but not all bottles were reactive. With S. marcescens (< 2 CFUs/mL), only 7 of the 10 sets were reactive. With C. perfringens (0.2 CFUs/mL), only 3 of 10 samples were detected in the anaerobic bottles. CONCLUSIONS: This study evaluates the use of the BacT/ALERT system for detection of bacteria in PLT pools. Overall, the BacT/ALERT detected all contaminated pooled PLTs when the single inoculated unit had a calculated or recovered concentration at least 3 CFUs per mL with 10 different species of bacteria. Low recovered concentrations (< or =2 CFUs/mL) were, in some cases, only detected with a two-bottle set.

Blood Platelets↗
PubMed2005

Decreased sepsis related to indwelling venous access devices coincident with implementation of universal leukoreduction of blood transfusions.

BACKGROUND: Randomized trials and animal models demonstrate that leukoreduction of transfusions can reduce the risk of post-operative infections. We performed a retrospective study of sepsis related to indwelling venous access devices (line related infections) before and after the July 2000 implementation of universal leukoreduction. METHODS: Line related infection data were collected from hospital infection control records for an 18-month period before and after July 2000. Transfusion histories were obtained from transfusion service records. RESULTS: Line related infections decreased in number from 150 to 98 (-35%) in transfused patients after implementation of universal leukoreduction, whereas line related infections increased from 41 to 45 (+10%) in nontransfused patients (p = 0.04). This corresponded to a decrease from 5.3 to 3.3 infections/10,000 patient days in transfused patients (p = 0.002). The infection rate remained stable in nontransfused patients at 1.5 infections/10,000 patient days both pre- and postimplementation of universal leukoreduction. Quantitatively similar decreases (33-45%) were seen in transfused surgical, medical and pediatric patients. CONCLUSIONS: A substantial and statistically significant decrease in line related infections occurred coincident with implementation of universal leukoreduction. These improved outcomes were observed in transfused but not nontransfused patients, across all clinical services, suggesting a causal relationship with universal leukoreduction.

Adolescent↗
PubMed2005

Removal of exogenous (spiked) and endogenous prion infectivity from red cells with a new prototype of leukoreduction filter.

BACKGROUND: Two recent probable cases of transmission of a variant of human Creutzfeldt-Jakob disease (vCJD) through blood transfusion suggest that the disease can be transmitted through transfusion of blood components from presymptomatic blood donors. In the absence of a preclinical screening test, removal of the infectious agent by processing is the only means by which risk to recipients of blood from donors with inapparent vCJD infections can be eliminated. STUDY DESIGN AND METHODS: In the endogenous infectivity study, a pool of 500 mL of whole blood was collected into CP2D anticoagulant from 263K-strain scrapie-infected hamsters, processed into 300 mL of red cells (RBCs), and then passed through a prion removal filter. Pre- and postfiltration samples were tested for PrP(sc) by Western blot and for infectivity by inoculation of healthy hamsters. In the exogenous (spiking) infectivity study, 30 mL of 10 percent (wt/vol) scrapie-infected brain homogenates was added to 270 mL of human RBCs and then filtered. Levels of PrP(sc) and infectivity were determined by Western blot and bioassay. RESULTS: In the endogenous infectivity study, the prefiltered RBCs transmitted disease to 6 of 43 animals, whereas the postfiltered RBCs did not transmit disease to any of 35 animals, and a barely visible prefiltration PrP(sc) Western blot signal was reduced below the level of detection in the postfiltration sample. In the exogenous (spike) study, infectivity was reduced by 3.7 log LD50 per mL, from 9.2 to 5.5 log LD50 per mL. CONCLUSION: The new filter was effective in removing both infectivity and PrP(sc) from RBCs. The use of this type of filter should reduce the risk of vCJD transmission through blood transfusion.

Animals↗
PubMed2006

Quality control of blood irradiation: determination T cells radiosensitivity to cobalt-60 gamma rays.

BACKGROUND: To identify the most appropriate dose for the prevention of transfusion-associated graft-versus-host disease, the radiosensitivity of T cells has been determined in blood bags irradiated with X-rays produced by a linear accelerator and gamma rays derived from the cesium-137 source of a specific irradiator. In this study, the influence of doses ranging from 500 to 2500 cGy was investigated on T cells isolated from red blood cell (RBC) units preserved with ADSOL and irradiated with a cobalt teletherapy unit. STUDY DESIGN AND METHODS: A thermal device consisting of acrylic and foam was constructed to store the blood bags during irradiation. Blood temperature was monitored with an automated data acquisition system. Dose distribution in the blood bags was analyzed based on isodose curves obtained with a polystyrene phantom constructed for this purpose. The influence of cobalt-60 gamma radiation on T cells was determined by limiting-dilution analysis, which measures clonable T cells. T-cell content of the mononuclear cell population plated was assessed by flow cytometry with a monoclonal antibody specific for CD3. RESULTS: Blood temperature ranged from 2 to 4.5 degrees C during irradiation. Dosimetry performed on the phantom showed a homogenous dose distribution when the phantom was irradiated with a parallel-opposite field. A radiation dose of 1500 cGy led to the inactivation of T cells by 4 log, but T-cell growth was observed in all experiments. At 2500 cGy, no T-cell growth was detected in any of the experiments and a greater than 5 log reduction in functional T cells was noted. CONCLUSION: The results showed that a dose of 2500 cGy completely inactivates T cells in RBC units irradiated with cobalt-60 source.

Adenine↗
PubMed2005

Prestorage leukoreduction and low-temperature filtration reduce hemolysis of stored red cell concentrates.

BACKGROUND: Universal prestorage leukoreduction in Canada created the perception that stored red cells (RBCs) are more hemolyzed than their unfiltered predecessors. A pool-split design tested the effects of leukoreduction on hemolysis of stored RBCs. STUDY DESIGN AND METHODS: Two ABO-matched units were pooled, divided, and then processed into leukoreduced (LR) and nonleukoreduced (NLR) units with the Pall LT-WB or RC-PL systems and sampled during standard processing and storage for testing of sterility, counts, hemolysis, and osmotic fragility. RESULTS: Room temperature (RT) filtration of 10 pairs of LT-WB-LR and -NLR units showed significantly different percentage of hemolysis (0.39%) and osmotic fragility (0.643%) at 42 days. Cold-stored and -filtered units (2 days at 4 degrees C before processing) were less hemolyzed, but showed a similar proportional decrease of hemolysis in LR units (0.13% vs. 0.25% at 42 days). RBCs from RC-PL systems showed the lowest hemolysis although there was a filtration effect (0.05% vs. 0.12%, 42 days). Osmotic fragility paralleled hemolysis. Segment samples gave inaccurate results. Two-day prefiltration cold storage reduced hemolysis from 0.36 to 0.07 percent (42 days, p < 0.001). RT-LR hemolysis became significantly higher by Day 10 and 4 degrees C LR by Day 12. NLR units showed hemolysis by Day 7. LR units filtered cold were less hemolyzed (p < 0.05) than RT-LR but osmotic fragility was unchanged. CONCLUSIONS: LR-RBCs prepared by any of three methods (LT-WB, RT or cold; RC-PL), filtered at 4 degrees C, were less hemolyzed during storage than nonfiltered concentrates: 4 degrees C leukoreduction is beneficial for RBCs and does not cause hemolysis or enhance fragility.

Blood Preservation↗
PubMed2005

Risk for postoperative infection after transfusion of white blood cell-filtered allogeneic or autologous blood components in orthopedic patients undergoing primary arthroplasty.

BACKGROUND: This study was designed to obtain data on the incidence of postoperative infection in patients undergoing elective orthopedic surgery and receiving white blood cell (WBC)-filtered blood components prepared according to current standards. STUDY DESIGN AND METHODS: A total of 308 consecutive orthopedic patients who opted for preoperative autologous blood donation (PAD) for primary unilateral hip and knee replacement surgery were enrolled in a prospective observational study of the incidence of postoperative infection. Patients with contraindications for PAD or with any infectious disease were not included in the study. To identify probably confounding factors, differences between patient groups were analyzed first. Identified factors, which differed between groups, and variables describing blood supply were further tested in uni- and multivariate logistic regression analysis for their independent influence on development of postoperative infection. Infection rates were compared on the basis of actual transfusion groups. RESULTS: Of the 308 study patients, 101 were not transfused, 85 received their PAD, 100 received allogeneic WBC-filtered red blood cells (RBCs), and 22 were given autologous RBCs and additionally allogeneic WBC-filtered RBCs. Overall the infection rate was 6.82 percent (21/308). Infection rates varied significantly between transfusion groups (no transfusion, 6.9%; autologous RBCs, 1.2%; allogeneic WBC-filtered RBCs, 12.0%; both transfusion types, 4.6%; p = 0.03). Allogeneic recipients showed significantly more infections compared to autologous recipients (p = 0.0053). Multivariate regression analysis confirmed transfusion of allogeneic WBC-filtered RBCs as an independent variable predicting postoperative infection (odds ratio, 23.65; confidence interval, 1.3-422.1; p = 0.01). CONCLUSION: Differences in postoperative infection rates between allogeneic and autologous recipients are still observable, although universal WBC filtration has been introduced into clinical practice.

Aged↗
PubMed2005

Blood donor white blood cell reduction filters as a source of human peripheral blood-derived endothelial progenitor cells.

BACKGROUND: Neovascularization in tumors, wounds, and sites of ischemic injury occurs by both angiogenesis (proliferation from existing vessels) and by vasculogenesis (differentiation into endothelial cells from circulating endothelial progenitor cells [EPCs]). EPCs can be obtained from marrow, from cord blood, or by ex vivo expansion of human peripheral blood (PB). The ease of obtaining human PB EPCs has led many recent studies to utilize PB EPCs. The ability to obtain large numbers of PB EPCs would greatly facilitate characterization to further our understanding of EPC biology and their application in cellular gene therapy. STUDY DESIGN AND METHODS: Peripheral blood mononuclear cells (PBMNCs) from whole blood or from the material obtained from white blood cell (WBC) reduction filters were isolated. The cells from both sources were then cultured separately under defined conditions to quantify EPC yield from each source. RESULTS: The yield of EPCs per million PBMNCs plated was approximately 3.5-fold higher from fresh PB. Because greater numbers of PBMNCs were obtained from each filter, however, the mean yield of EPCs from one filter versus fresh blood was 5.4 million versus 0.4 million, respectively (approx. 14-fold increased yield). CONCLUSION: The use of WBC reduction filters provides a safe, inexpensive, and readily available source for large numbers of PBMNCs from which culture-expanded EPCs can be generated for further study.

Blood Donors↗