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Background and Objectives Since 2001, all blood components in Germany must be leucocyte depleted. Recently, a new method for quality control of depletion was introduced. Our study aimed at the validation of the method for routine use in apheresis platelet concentrates. Materials and Methods We compared the new ADAM-rWBC device with manual counting in the Nageotte chamber and flow cytometry, two standard methods, by measuring residual leucocytes in 40 units of apheresis platelet concentrates and in six geometrical dilution series. Results Cell counts of residual leucocytes in the 40 units were below 106 cells per component with all methods, although mean cell counts were approximately 5 and 6 times higher in flow cytometry and ADAM-rWBC, respectively, compared to the Nageotte chamber. No unit with

In most cases, the amount of hematopoietic stem and progenitor cells (HSPCs) in a single cord blood (CB) unit is not sufficient for allogenic transplantation of adults. Therefore, two CB units are usually required. The ex vivo expansion of HSPCs from CB in coculture with mesenchymal stroma cells (MSCs) might be an alternative. It was investigated, whether bone marrow-derived MSCs, which have to be obtained in an invasive procedure, introduce a further donor and increases the risk of transmissible infectious diseases for the patient can be replaced by MSCs from amnion, chorion, Wharton's jelly, amniotic fluid, and CB, which can be isolated from placental tissue which is readily available when CB is sampled. In a two-step ex vivo coculture mononuclear cells from cryopreserved CB were cultured with different MSC-feederlayers in a medium supplemented with cytokines (stem cell factor, thrombopoietin [TPO], and granulocyte colony-stimulating factor). Expansion rates were analyzed as well, by long-term culture-initiating cell (LTC-IC) and colony-forming unit (CFU) assays, as by measuring CD34(+)- and CD45(+)-cells. Due to the comparably low number of 5×10(2) to 1×10(4) CD34(+)-cells per cm(2) MSC-monolayer, we observed comparably high expansion rates from 80 to 391,000 for CFU, 70 to 313,000 for CD34(+)-, and 200 to 352,000 for CD45(+)-cells. Expansion of LTC-IC was partly observed. Compared to the literature, we found a better expansion rate of CD34(+)-cells with MSCs from all different sources. This is probably due to the comparably low number of 5×10(2) to 1×10 CD34(+)-cells per cm(2) MSC-monolayer we used. Comparably, high expansion rates were observed from 80 to 391,000 for CFUs, 70 to 313,000 for CD34(+)-, and 200 to 352,000 for CD45(+)-cells. However, the expansion of CD34(+)-cells was significantly more effective with MSCs from bone marrow compared to MSCs from amnion, chorion, and Wharton's jelly. The comparison of MSCs from bone marrow with MSCs from CB and amniotic fluid showed no significant difference. We conclude that MSCs from placental tissues might be useful in the expansion of HSPCs, at least if low numbers of CD34(+)-cells per cm(2) MSC-monolayer and a high TPO concentration are implemented in the expansion culture.

BACKGROUND: The objective was to investigate potential risks for apheresis donors associated with a triple-plateletpheresis (TP) program. STUDY DESIGN AND METHODS: Eleven hemapheresis centers randomly assigned 411 repeat donors (ratio, 1:1.2) to either double plateletpheresis (DP; 185 donors) or TP (226 donors) with a platelet (PLT) target content of at least 5.0 × 1011 PLTs/DP and at least 7.5 × 1011 PLTs/TP. The primary endpoint was procedure-related postapheresis PLT count of at least 150 × 109/L (probability, ≥98%). Secondary endpoints were apheresis characteristics and donor adverse reactions. RESULTS: In 6 of 1133 DPs (0.5%) in 4 of 185 donors (2.2%) and in 20 of 1020 TPs (2.0%) in 14 of 226 donors (6.2%), postapheresis PLT counts were below 150 × 109/L. There were marginal but significant differences in collection efficiency (DP, 69.2 ± 9.1%; TP, 70.9 ± 9.0%; p ≤ 0.0001) and collection rate (DP, 10.4 × 109 ± 2.3 × 109 PLTs/min; TP, 10.8 × 109 ± 2.3 × 109 PLTs/min; p ≤ 0.005). The PLT yields were 5.9 × 1011 ± 0.8 × 1011 PLTs for DP and 8.3 × 1011 ± 0.9 × 1011 PLT for TP (p ≤ 0.0001) at processing times of 59 ± 13 minutes (DP) versus 80 ± 16 minutes (TP; p ≤ 0.0001). Significant PLT recruitment (1.10 ± 0.14 vs. 1.20 ± 0.23; p

BACKGROUND: The standard mononuclear cell (MNC) program of the COM.TEC device (Fresenius HemoCare GmbH) showed excellent collection efficiency of CD14+ monocytes. A major disadvantage was high content of residual cells in MNC harvests, which could influence dendritic cell (DC) culture. STUDY DESIGN AND METHODS: The autoMNC program (COM.TEC) was compared with the standard MNC program (n = 12). Additionally, two cycle volumes (300 mL vs. 450 mL, n = 19) were compared (standard MNC program). Samples were assayed for white blood cells (WBCs), red blood cells (RBCs), granulocytes (PMNs), hematocrit, and platelets (PLTs) on an automated blood cell counter (Sysmex K 4500, TAO Medical). CD14+ cells were analyzed by flow cytometry (FACSCalibur, BD). RESULTS: The autoMNC program produced 1.33 × 109 ± 0.36 × 109 CD14+ cells, 5.60 × 1011 ± 0.97 × 1011 PLTs, and 1.43 × 1011 ± 0.37 × 1011 RBCs. Compared to the standard MNC program, significantly higher PLT yields but lower RBC yields and product volume were harvested. Increasing the CV from 300 to 450 mL dropped the product volume, residual PLTs, and RBCs significantly, whereas WBC and monocyte yields did not change. The WBC predonation counts of donors correlated significantly with monocyte yields. CONCLUSIONS: The autoMNC program reduced the buffy coat (BC) volume and RBC yields in products compared to the standard MNC program. Increasing the CV (standard MNC program) reduced residual PLTs, RBCs, and the BC volume of MNC harvests. The donor WBC predonation count was a good predictor for the monocyte yield of products.

Background The aim of this study was to investigate the effect of gamma irradiation with 30 Gy on the coagulation system in leukoreduced fresh-frozen plasma (FFP). Study design and methods In 74 FFP units that had been stored for 352 +/- 103 days below -30 degrees C, the following variables were determined in parallel in an irradiated and not irradiated half: prothrombin time (PT); activated partial thromboplastin time (APTT); thrombin time; antithrombin III; protein C; protein S; von Willebrand factor antigen; ristocetin cofactor; plasminogen-alpha(2)-antiplasmin; the coagulation factors fibrinogen, factor (F)II, FV, FVII, VIII, F IX, FX, FXI, FXII, FXIII, and activated factor XII (FXIIa); D-dimer; fibrin monomer; thrombin-antithrombin complex; prothrombin fragment 1 + 2 (F1+2); plasmin-alpha(2)-antiplasmin complexes (PAPs); and platelet factor 4. The FVII activity ratio was assayed to quantify activation of FVII. Results Irradiation with 30 Gy resulted in a reduction of APTT (35.0 +/- 4.1 sec vs. 34.4 +/- 4.1 sec; p = 0.00000006) and PT (89.8 +/- 8.2% vs. 90.7 +/- 8.0%; p = 0.002) and a significant increase of the activities of the coagulation factors FII, FV, FVII, F IX, FX, and FXII. FVIII activity decreased from 118 +/- 31 to 116 +/- 32 percent (p = 0.02). Activation of the coagulation system was shown by an increase in the FVII activity ratio (1.19 +/- 0.29 vs. 1.31 +/- 0.34; p = 0.0000001), FXIIa (0.81 +/- 0.50 ng/mL vs. 0.90 +/- 0.51 ng/mL; p = 0.006), and F1+2 (1.19 +/- 0.20 nmol/L vs. 1.24 +/- 0.20 nmol/L; p = 0.000005) after irradiation with 30 Gy, whereas an increase of PAP (16.2 +/- 11.5 ng/mL vs. 20.2 +/- 12.0 ng/mL; p = 0.0004) demonstrated activation of the fibrinolytic system. No negative influence of irradiation with 30 Gy on inhibitors of coagulation was observed. Conclusion Gamma irradiation of leukoreduced FFPs with 30 Gy results in a significant but very weak activation of the coagulation and fibrinolytic system in FFPs.

BACKGROUND: The Fresenius COM.TEC cell separator is a new device for producing white cell concentrates (WBCs) and leukoreduced single-donor platelet concentrates (SDPs) and performing therapeutic cytapheresis and plasmapheresis that might replace the Fresenius systems AS104 and AS.TEC 204. This novel system's performance was evaluated for producing leukoreduced SDPs. STUDY DESIGN AND METHODS: In an investigational phase, each of 200 donors underwent plateletpheresis with the AS.TEC 204 and the COM.TEC systems. The collection efficiency (CE) and WBC contamination of the different techniques were compared. After some hard- and software modifications, the system was evaluated in an additional 800 procedures in the confirmatory phase. RESULTS: In the investigational phase, the CE of the COM.TEC device was increased significantly in comparison to the AS.TEC 204 device's CE (by 45 ± 32% when collecting 1 unit of platelets [PLTs] and 1 unit of fresh-frozen plasma and by 43 ± 42% when collecting only 1 unit of PLTs). Although all AS.TEC products proved to be leukoreduced, 2 percent of the COM.TEC procedures led to PLT concentrates containing more than 1 × 106 WBCs. In the confirmatory phase, all 1300 products from 800 COM.TEC procedures proved to be leukoreduced. Furthermore, the CE increased significantly from 53.5 ± 4.6 percent in the investigational phase to 55.5 ± 4.9 percent (p

Recently, two prospective randomized multicenter studies on hypoproliferative thrombocytopenia compared prophylactic transfusion of platelet concentrates below a threshold of 10 000/μL with therapeutic platelet transfusions only, administered at onset of hemorrhages other than mucocutaneous bleeding (1, 2). In the patient group with only therapeutic platelet transfusion, a significant increase in the incidence of grade 2, 3 or 4 (World Health Organization [WHO] bleeding scale) hemorrhages was observed. This applied to both patients with acute myeloid leukemia (AML) and patients after autologous stem cell transplantation. However, since the most severe form of hemorrhage—intracerebral hemorrhage—occurred only in AML patients, Wandt et al. (1) regarded the available evidence as being sufficient to adopt a therapeutic-only platelet transfusion strategy in adult, clinically stable patients after autologous stem cell transplantation. However, this strategy requires monitoring at close intervals for signs of hemorrhage which can be realized during clinical studies, but not in a standard hospital setting. In addition, the immediate availability of platelet concentrates in the event of hemorrhage cannot be guaranteed at all times, even in tertiary care hospitals. Moreover, platelet concentrates with reduced platelet content and pathogen-inactivated platelet concentrates are commercially available in German-speaking countries. Prospective studies evaluating these types of platelet concentrates observed sporadic cases of intracranial hemorrhage even when platelet transfusions are applied prophylactically. To pursue a therapeutic-only platelet transfusion strategy using these types of platelet concentrates would expose patients to unpredictable risks. In the best interest of the safety of patients suffering from hypoproliferative thrombocytopenia, we firmly reject the position of Wandt et al. (3) and agree with Professor Slichter from Seattle who wrote in a 2013 editorial of the New England Journal of Medicine (4),′In my opinion, the reduction in the use of platelet transfusions does not justify subjecting patients to the increased bleeding risks associated with a therapeutic-only platelet-transfusion strategy in any category of patients with hypoproliferative thrombocytopenia′.

BACKGROUND: The aim of this study was to investigate the occurrence of hypotension in the 24-hour period after preoperative autologous blood donation (PABD) in patients with and without hypertension. STUDY DESIGN AND METHODS: In 20 patients, 24-hour ambulatory blood pressure monitoring (ABPM) was performed before PABD was started and on every donation day in two repeated phlebotomies. RESULTS: Seven patients had no hypertension and 11 patients had hypertension. In 2 additional patients, hypertension was diagnosed during the study. Overall, the mean systolic BP (SBP) decreased from 131 ± 15 mmHg before donation to 128 ± 13 and 127 ± 10 mmHg after Donations 1 and 2; the corresponding values for the diastolic BP (DBP) were 77 ± 9, 75 ± 9, and 73 ± 7 mmHg, both without significant differences between the groups with and without hypertension. In single patients, substantial decreases of BP occurred, especially during the night. Two patients with and 2 without hypertension showed a nightly decrease in SBP and DBP of more than 10 percent (in 1 of these patients, more than 20%). Concerning diurnal BP variability, 1 patient with and 1 without hypertension, the latter showing a nightly decrease of SBP and DBP of more than 10 percent, also changed to the pattern of a nightly “extreme dipper” after PABD. CONCLUSION: In 25 percent of the patients, changes of BP were observed during the 24-hour period after PABD, especially during the night, which are known to be associated with an increased risk of cerebral or myocardial ischemia. Whether those changes of BP lead to major morbidity or mortality requires further investigation.

BACKGROUND: Only a few systematic studies have examined the effect of variable produced small platelet (PLT) aliquots on PLT function before transfusion to neonates. Although neonatal transfusion could be critical, no standardization of production or systematic quality controls have been introduced so far. STUDY DESIGN AND METHODS: PLT split products were prepared in three different ways (30- or 60-mL bag, syringe aliquot) at different times from the parent unit (Days 1-4) and stored for 2 or 4 hours. The measures of PLT function include pH, lactate, P-selectin expression, and cytokines (β-thromboglobulin [β-TG], PLT-derived growth factor AB [PDGF-AB]). Additionally, syringe passage (0.5 mL/min) was assessed. RESULTS: High product variability of PLT content was found (40% deviation of PLT content from programmed target, 13%-19% PLT loss by product distribution), which resulted in PLT concentrations of split units between 0.94 × 109 and 1.66 × 109 PLTs per mL. Different gas transfer rates (pCO2) of PLT containers caused different pH values of the product (Trima 7.47 ± 0.09 vs. COM.TEC 7.33 ± 0.08; p

Background and objectives Lowering the plasma content in single-donor platelet (PLT) concentrates well below 30% implies the need to collect platelets at very high concentrations. Trima Accel (TA) is validated for collection below 4000 x 10(3) PLTs/microl. We evaluated its performance at 5000 x 10(3) PLTs/microl. Materials and methods Twenty blood donors underwent apheresis with TA twice collecting either a hyperconcentrated or a standard single-donor platelet concentrate with a target platelet concentration of 5000 or 1200 x 10(3) PLTs/microl, respectively. We analysed the collection efficiency, the collection rate and the quality of the collected by-plasma. Results We collected 20 hyperconcentrated and 20 standard units containing 2.56 +/- 0.5 and 3.39 +/- 0.4 x 10(11) PLTs at a concentration of 4518 +/- 978 and 1374 +/- 166 x 10(3) PLTs/microl in 45 +/- 8 and 39 +/- 6 min resulting in a collection efficiency of 47.5 +/- 10.0 and 70.7 +/- 7.9% and a collection rate of 5.9 +/- 1.4 and 8.8 +/- 1.5 x 10(9) PLTs/min, respectively (all results expressed as mean +/- standard deviation). The collected by-plasma showed a very high grade of cell purity and a satisfactory recovery of the clotting factors. Conclusion Although TA is a suitable device for PLT collection at very high concentrations, improvements are desirable to further increase the productivity above its currently validated upper collect concentration limit.

BACKGROUND: Rare clinical conditions cause the need for washed platelet (PLT) concentrates (PCs). Saline-washed PCs can only be stored shortly, however, owing to lack of substrates for PLT metabolism. New PLT additive solutions (PASs) contain such substrates and might be used alternatively. The in vitro quality of apheresis PCs washed with Composol-PS or modified PAS-III (PAS-IIIM) stored up to 48 hours after wash was compared. STUDY DESIGN and METHODS: Twelve blood donors underwent two apheresis procedures (A and B) collecting 6.0 × 1011 PLTs in 500 mL of plasma with a least 2 weeks in between. The PCs collected by Apheresis A were stored for 3 days and then split in two equal units before washing with Composol-PS or PAS-IIIM. The PCs collected by Apheresis B were split after collection. One unit was released for transfusion and 1 unit was stored unwashed up to Day 6 and used as reference unit. In vitro testing was performed before and after washing as well as 24 and 48 hours after wash. RESULTS: After 48 hours of postwash storage, the units washed with either PAS showed acceptable results for hypotonic shock response (HSR), P-selectin expression, and pH, whereas PLT aggregability was significantly impaired. Throughout the storage, unwashed units showed better in vitro quality. HSR and P-selectin expression were similar before and immediately after the washing procedure. CONCLUSION: Based on these in vitro results, 48-hour postwash storage of washed PCs with the two PASs seems to be feasible. In vivo recovery studies, however, must confirm this finding in the future.

BACKGROUND: Standard plateletpheresis techniques and effects on platelet (PLT) donors were investigated to provide an informative basis for advancement of apheresis software. STUDY DESIGN AND METHODS: Three paired groups with 33 male and 22 female blood donors were prospectively investigated by analyzing blood counts of donors and products. Four apheresis platforms, the COBE Spectra LRS and the Trima v4 (Gambro BCT) and the AS.TEC204 and the COM.TEC (Fresenius Hemocare), were compared. Deviations of the collected from programmed PLT targets and donor PLT recruitment were calculated for single-unit PLT concentrates (SU-PCs; 3 × 1011 PLTs) and double-unit PLT concentrates (DU-PCs; 6 × 1011 PLTs). RESULTS: Regarding SU-PCs, the productivity of the COM.TEC machine was superior to the AS.TEC204 machine, because of shorter processing time (54 min vs. 67 min) and higher yields (2.90 × 1011 PLTs vs. 2.75 × 1011 PLTs). Compared to the Spectra machine, the Trima v4 machine showed higher collection efficiencies (CEs) and shorter processing time and complied better with the programmed target (SU-PCs, 3.24 × 1011 PLTs vs. 3.70 × 1011 PLTs; DU-PCs, 6.87 × 1011 PLTs vs. 7.56 × 1011 PLTs). Harvests of the Spectra machine (DU-PCs) exceeded the target by 40 percent, which resulted in high PLT loss for donors. A longer processing time resulted in some higher CEs (SU-PCs, 53%; DU-PCs, 58%), which could contribute to this result. PLT recruitment compensated PLT loss to some extent. CONCLUSION: The major finding was that the newer devices (COM.TEC and Trima) gave more predictable yields than the older devices (AS.TEC204 and Spectra) and resulted in lower PLT deficit. PLT software should be improved to minimize relevant variations of collected yields regarding the programmed target.

Background and objectives No data are currently available on the quality of washed prestorage leucocyte-depleted red blood cell concentrates (RCCs). Materials and methods Five groups of RCCs stored in additive solution (SAG-M) were washed. The groups differed in the age of RCCs (2-5 days or 11-15 days), the temperature during the washing procedure and a 6-h storage period (4 degrees C or room temperature) and the washing solution (saline, SAG-M or 5% albumin). We measured ATP, 2,3-diphosphoglycerate (2,3-DPG), haemolysis, blood cell count, Na(+), K(+), pH, pO(2), pCO(2) and lactate, before and after the washing procedure and hourly during the 6-h postwash storage period. Results The erythrocyte ATP content increased by 2-13%, relative to the baseline value, during the washing procedure. The 2,3-DPG level decreased by 15-35% in 2-6-day-old RCCs and by 30-40% in 11-15-day-old RCCs (relative to baseline values) during the washing procedure. In RCCs that were washed and stored at room temperature, and in 2-week-old RCCs, a further decrease in 2,3-DPG of up to 40%, relative to the baseline value, was observed during the 6-h postwash time-period. Conclusions Washing of RCCs stored in SAG-M results in a considerable, significant loss of erythrocyte 2,3-DPG, especially in older RCCs. This loss increases in during a 6-h storage period postwash, even at 4 degrees C. This loss of erythrocyte quality might well outweigh the benefits of washed SAG-M RCCs during massive transfusion in neonates.

BACKGROUND: In cancer and transplantation therapy apheresis devices and software of optimum standards are required for the collection of high cell yields with high purity of the desired cell fraction. STUDY DESIGN AND METHODS: In a paired study, 15 healthy blood donors underwent four 10-L leukapheresis procedures (197 ± 33 min) with an inlet blood flow rate of 60 mL per minute by use of two different MNC program settings of the COBE Spectra (Gambro BCT) and the AS.TEC 204 (Fresenius Hemocare) cell separators. RESULTS: Use of the standard MNC program of both apheresis devices resulted in significantly higher (p < 0.01) collection efficiencies of CD14+ monocytes, CD3+ cells, CD4+ cells, CD8+ T cells, CD16+ CD56+ natural killer (NK) cells, and residual PLTs (p < 0.001), owing to higher centrifuge speed. The mean MNC purity of all components was more than 90 percent. By use of stan-dard programs of either device, significant correlations (p < 0.01) between donor monocytes and preleukapheresis NK cell counts and the corresponding component cell yields were found. CONCLUSION: Compared to the program modifications with lower centrifuge velocities the standard MNC programs were significantly more efficient regarding CD14+, CD3+, and CD16+ CD56+ cells. Enhanced centrifuge speed and inlet blood flow rate in MNC programs resulted in higher, similar composed MNC concentrations of the products.

Background and Objectives Aside from new software the blood cell separator TRIMA (GambroBCT) also received a newly designed separation chamber offering a novel single stage separation technology, called Trima Accel. We evaluated this new system focusing on productivity and donor comfort by comparing it to the previous version (Trima version 4) in collecting single-donor platelet concentrates (SD-PCs) and plasma. Materials and Methods Each of 20 donors underwent platelet apheresis using both devices. We compared the collection efficiency (CE), the collections rate (CR), the volume of the collected plasma and the residual leukocytes. Furthermore we compared donor comfort in terms of duration of the donation, flow of citrate back to the donor and platelet and whit blood cell (WBC) loss. Results While the number of collected platelets and the platelet concentration did not differ significantly between both techniques the time of the procedure was reduced by 15.6% with Trima Accel. This results in an increase of the CR and CE of 25% and 15% respectively when using Trima Accel. Log normal probability plotting of WBC counts showed that both techniques complied with the European and the US leukoreduction guidelines. The mean flow of ACDA to the donor perminute and per litre blood volume was also reduced by 20%. Conclusion These data show that the Trima Accel represents a further improvement in apheresis platelet production with a better productivity and donor comfort, especially regarding the mean flow of ACDA to the donor.

The properties of red blood cell (RBC) concentrates stored in different additive solutions have been previously examined under laboratory conditions at the end of shelf-life. However, whether these data are representative for RBC units used in clinical practice has not been shown. Therefore, we examined 164 RBC units from six manufacturers outdated after clinical usage in a hospital-based transfusion service for cellular content, hemolysis, adenosin triphosphate, 2,3-DPG, pH, oxygen saturation and levels of beta-thromboglobulin and proinflammatory cytokines interleukin (IL) 1beta (IL-1), IL-6, IL-8 and tumor necrosis factor alpha (TNFalpha). Results were correlated with the number of interruptions of recommended storage conditions and with different manufacturers. TNFalpha and IL-8 levels in the supernatant of RBC concentrates showed a weak correlation with the number of interruptions of recommended storage conditions (TNFalpha: r = 0.25, P < 0.01; IL-8: r = 0.20, P < 0.01) for the whole series. We detected no significant correlation between hemolysis and interruptions of recommended storage conditions or any of the remaining studied parameters. However, we found significant differences between RBC concentrates supplied by different manufacturers with respect to cellular content and most of the studied parameters. RBC concentrates containing SAG-M from one single manufacturer had higher in vitro hemolysis at the end of shelf-life compared to all other manufacturers (P < 0.05). We conclude from our data that interruptions of optimal conditions for storage of red cell components during cross-match testing and transport in our setting play a minor role for in vitro properties of RBC units at the end of shelf-life. The influence of processes of production, storage and/or transport until entry of RBC units into our blood component depot seems to be much more important for final product quality at the end of shelf-life than subsequent events.

BACKGROUND: Monocytes collected by leukapheresis are increasingly used for dendritic cell (DC) culture in cell factories suitable for DC vaccination in cancer. STUDY DESIGN AND METHODS: Using modified MNC programs on two apheresis systems (Cobe Spectra and Fresenius AS.TEC204), leukapheresis components collected from 84 patients with metastatic malignant melanoma and from 31 healthy male donors were investigated. MNCs, monocytes, RBCs, and platelets (PLTs) in donors and components were analyzed by cell counters, WBC differential counts, and flow cytometry. RESULTS: In 5-L collections, Astec showed better results regarding monocyte collection rates (11.0 vs. 7.4 × 106/min, p = 0.04) and efficiencies (collection efficiency, 51.9 vs. 31.9%; p < 0.001). Both devices resulted in monocyte yields at an average of 1 × 109 (donors) and 2.5 × 109 (patients), whereas Astec components contained high residual RBCs. Compared to components with low residual PLTs, high PLT concentration resulted in higher monocyte loss (48 vs. 20%, p < 0.0001) before DC culture. CONCLUSION: The Astec is more efficient in 5-L MNC collections compared to the Spectra. Components with high residual PLTs result in high MNC loss by purification procedures. Thus, optimizing MNC programs is essential to obtain components with high MNC yields and low residual cells as prerequisite for high DC yields.

BACKGROUND: A new cell separator (COM.TEC, Fresenius) was recently developed aimed at efficient collection of WBC-reduced single-donor PLT concentrates (SDPs). STUDY DESIGN AND METHODS: Five German centers collected 554 WBC-reduced SDPs with help of the COM.TEC cell separator. Two multicenter cell counting studies were performed at the beginning and at the end of the study to document uniform counting results among the participating centers. RESULTS: A total of 441 (79.6%) PLT collections were included in the study according to the protocol. A total of 342 single-dose and 99 double-dose SDPs were collected. For single-dose SDPs, an average blood volume of 2826 ± 409 mL was processed in a donation time of 55 ± 11 minutes. Mean PLT yield of these products was 3.11 × 1011± 0.40 × 1011 and the WBC contamination was 0.11 × 106± 0.20 × 106. For double-dose SDPs (PLT count, 5.29 ± 0.93 × 1011), 3943 ± 639 mL was processed. The average difference between the target and the collected PLT concentration was −2.8 ± 12.0 percent for single-dose SDPs and −1.8 ± 9.5 for double-dose SDPs, respectively. The collection efficiency was 53.7 ± 5.8 percent for single-dose SDPs and 58.2 ± 6.2 percent for double-dose SDPs. If all results of each sample from the counting study were set to unity (to the mean over all centers), most PLT determinations were very similar to the mean, for example, near or 1 if set to unity. CONCLUSION: The COM.TEC machine makes it possible to obtain WBC-reduced SDPs that comply with current standards.

Background and objectives Regulatory T cells (Tregs) and other T-cell subsets are of importance in the setting of autologous and allogeneic stem cell transplantations. We conducted a study to assess the content of peripheral blood stem cell concentrates and related apheresis parameters in the autologous and allogeneic setting. Material and methods We characterized 53 donors, patients and peripheral blood stem cell concentrates (PBSC) regarding the content of CD45(+) cells, lymphocytes, CD3(+) cells, CD3(+) CD4(+) T cells, CD3(+) CD4(+) CD25(+) T cells, CD3(+) CD4(+) CD25(+) CD127(low/negative) Tregs and CD34(+) cells and calculated cell yields, recruitment factors and collection efficiency for all cell types. We compared allogeneic data with autologous data. Results Autologous PBSC show significantly lower concentrations of T-cell subsets compared to allogeneic PBSC (17,112/μl CD4(+), 14,858/μl CD4(+) CD25(+) and 1579/μl CD3(+) CD4(+) CD25(+) CD127(low/negative) Tregs in autologous compared to 65,539/μl CD4(+), 44,208(+) /μl CD4(+) CD25(+) and 5040/μl CD3(+) CD4(+) CD25(+) CD127(low/negative) Tregs in allogeneic PBSC, respectively), in contrast to CD34(+) concentrations (5342/μl CD34(+) in autologous compared to 2367/μl CD34(+) in allogeneic PBSC, respectively). Accordantly, all T-cell yields are lower in the autologous setting compared to allogeneic PBSC. However, recruitment factor and collection efficiency of all cell types are higher in autologous compared to allogeneic PBSC, but not all parameters differ significantly when groups are compared. Conclusion T-cell subsets and especially Tregs are a substantial part of PBSC transplantation, as considerable recruitment during apheresis occurs. In large volume apheresis, the collection efficiency of Treg is comparable to that of CD34(+) cells, while recruitment factors are even higher.

BACKGROUND: A cell separator (Spectra, Gambro BCT) with an integrated leukoreduction system (LRS) for producing WBC-reduced single-donor platelet concentrates has been shown to result in a slightly reduced collection efficiency as compared to the former Spectra system without LRS. A novel modified system for improved collection efficiencies (LRS Turbo, Gambro BCT) was evaluated. STUDY DESIGN AND METHODS: Each of 37 donors underwent plateletpheresis using the LRS Turbo (LRS-T) and the standard LRS (LRS) of the Spectra cell separator. The collection efficiency and WBC contamination of the different techniques were compared. Platelets were counted automatically and WBCs were counted by using one or two full grids of a Nageotte chamber. RESULTS: The preseparation and postseparation numbers of RBCs, WBCs, and platelets, as well as the number of collected platelets, did not differ for the two techniques. In the LRS-T separations, the collection efficiency was 112 percent of that in the LRS procedures. Median residual WBCs in the platelet components were 0.0256 × 106 per LRS-T procedure and 0.0253 × 106 per LRS procedure. The purity of the LRS-T components was not less than that of the standard LRS components, whereas the collection efficiency of the LRS-T was significantly greater, 44.9 percent versus 40.7 percent. CONCLUSIONS: The LRS-T procedures produced platelet concentrates with WBC-reduction capacity that is comparable to that obtained with the standard LRS procedures, which have previously been described as satisfying the most stringent criteria for WBC-reduced platelets. The new technique significantly improved the collection efficiency of the plateletpheresis procedure.

BACKGROUND: Compensatory RBC production during repeated preoperative autologous blood donation (PABD) shows marked interindividual variability. This study was performed to reveal variables that might be useful to predict the amount of the erythropoietic response to PABD in an individual patient who was not iron deficient. STUDY DESIGN AND METHODS: In a retrospective study, 104 adult patients, 48 women and 56 men (mean age, 59.9 years; range, 18-82 years) who donated 3 units (450 mL) of autologous blood at weekly intervals for major surgery were investigated. Blood counts, ferritin, and net preoperative RBC production (net RBC production) were determined in all patients, and soluble transferrin receptor and endogenous levels of EPO, SCF, and IL-1β were measured in 63 patients. Multiple linear regression analysis was used to determine whether the variance of net RBC production was attributable to baseline values of these variables. RESULTS: Net RBC production was not different in patients who received oral iron and patients who did not (384 ± 222 mL vs. 356 ± 158 mL). In both groups, the same two variables consistently showed a significant relationship to net RBC production: the length of the period between the third donation and the last visit was positively related (p = 0.00001 vs. p = 0.0002) and the Hct at baseline was negatively related (p = 0.0002 vs. p = 0.02) with net RBC production. The proportion of variance in net RBC production that was attributable to these two variables was 48.1 percent (r2 = 0.481) and 34.9 percent (r2 = 0.349), respectively. CONCLUSION: RBC production after PABD increases with increasing interval from last donation to surgery. This suggests that the interval from last donation to surgery should be maximized. This can be achieved by organizational measures in combination with the preparation of RBC concentrates in additive solution with a maximum shelf life.

BACKGROUND: A new software version of a cell separator (AS TEC 204, Fresenius) providing WBC-reduced single-donor plateletpheresis concentrates was tested. STUDY DESIGN AND METHODS: Dual-needle apheresis procedures (n = 621) were performed in three centers, using either fixed interface positioning (FIP) or periodic alternating interface positioning (PAIP). The other separation parameters (e.g., anticoagulant:whole-blood ratio, and blood flow) were set individually. All platelet concentrates were evaluated for platelet yields and contaminating WBCs. RESULTS: The introduction of the PAIP resulted in a significant (p

Background This study was performed to evaluate the capacity of oral and intravenous (i.v.) iron administration during autologous blood donation (ABD) to improve the efficacy of ABD and to prevent the need for allogeneic blood transfusion in patients without iron deficiency who are undergoing major elective surgery for which a minimum of 3 autologous units have been ordered. Study design and methods One hundred twenty-three patients were enrolled in an open-labeled, randomized, controlled trial and assigned to three treatment groups: patients in Group 1 received 3 x 100 mg of Fe2+ per day given orally for 5 weeks before operation; patients in Group 2 received 200 mg of Fe3+ given intravenously after each donation combined with initial i.v. iron supplementation in patients with hemoglobin under 15 g per dL; and patients in Group 3 were in the control group that received no iron medication. A modest ABD program involving weekly phlebotomy and threshold hemoglobin values for donation of 11.5 g per dL in women and 12.0 g per dL in men was performed. Results Ninety patients, 15 women and 15 men in each of the three groups, completed the study. The mean net red cell production during ABD was no higher (p>0.2) in the iron-treated groups (Group 1: 473 +/- 178 mL; Group 2: 436 +/- 170 mL; Group 3 (controls): 397 +/- 174 mL). The mean number of autologous units donated per patient did not differ (p>0.7) among the groups (Group 1: 3.1 +/- 0.6; Group 2: 2.9 +/- 0.7; Group 3: 3.0 +/- 0.7). The proportion of patients who needed allogeneic blood transfusion showed no significant (p>0.4) advantage for iron treatment, (Group 1: 7%; Group 2: 20%; Group 3: 10%). Conclusion In non-iron-deficient patients undergoing modest ABD without erythropoietin therapy, neither oral nor i.v. application of iron during the preoperative period enhances the success of preoperative ABD.

BACKGROUND: Quality assurance of blood transfusion covers institutions, personnel, and procedures involved in preparing, issuing, and using blood components. The accuracy of data related to blood component transfusions is a tool for quality control in the transfusion service. STUDY DESIGN AND METHODS: A study of the accuracy of data records of the transfusion service at the University Hospital of Erlangen, Germany, between June 1994 and May 1996 was carried out. All returned blood component transfusion report forms were examined for discrepancies between primary data records and clinical transfusion reports. RESULSTS: Blood components (n = 49,224) from allogeneic and autologous donations, packed red cells, fresh-frozen plasma, and platelet components that had been issued for transfusion were included in this evaluation. For 27.3 percent of all components issued, no transfusion report was returned to the blood bank. For the remaining 35,786 units, errors were found in 3.8 percent of the records. For 1.24 percent of all components, discrepant information related to the recipient's identity or the component's status was found; this affected the feasibility of lookback or traceback searches. CONCLUSION: A remarkably high frequency of discrepancies exists between computerized blood bank records and the information recorded on returned blood transfusion forms. The processes of data acquisition and entry must be included in quality assurance efforts in transfusion medicine.

Background: The consequences of the new edition of the ‘German Guidelines for Blood Grouping and Blood Transfusion (Haemotherapy)’ for aptitude tests of haemapheresis donors were studied. Material and Methods:During a 19-month period the reasons for exclusion and deferral of first-time donors were retrospectively evaluated, and the resulting exclusion and deferral rates in accordance with the old and the new edition of the German guidelines were calculated. Results:From January 1995 to July 1996 979 first-time donors for haemapheresis donations were tested. Based on the old edition of the guidelines, 4.6% of the first-time donors were excluded and 16.8% were temporarily deferred after taking the health history. From 770 persons blood and urine samples and ECGs were taken. Due to pathological results 5% of the first-time donors were excluded and 13.5% were temporarily deferred. Using the new edititon of the guidelines, 12.5% of the first-time donors would have been deferred after taking the health history and 7.9% because of pathological laboratory tests. The new edition of the guidelines did not change the exclusion rate of first-time donors. Conclusion: The new edition of the ‘German Guidelines for Blood Grouping and Blood Transfusion (Haemotherapy)’ reduces the deferral rate of voluntary first-time donors. Nevertheless, the deferral rate of first-time donors in Germany exceeds the deferral rates reported from other countries.

BACKGROUND AND OBJECTIVES Because of widespread use of leukocyte reduction in platelet concentrates (PCs) and the need to store such concentrates, we investigated the effects of leukocyte depletion on the quality of stored PCs. MATERIALS AND METHODS Ten double-sized PCs were divided into 2 equal units which were tested simultaneously. One half was stored for 5 days after filtration through a polyester filter, the other one was stored unfiltered. RESULTS The volume of the 10 "oversized' PCs was 483 +/- 40 ml (mean +/- standard deviation) and they contained 5.9 +/- 1.5 x 10(11) platelets and 80 +/- 23 x 10(6) leukocytes. Filtration significantly reduced the leukocyte concentration (168 +/- 56/microliter before, 6 +/- 4 /microliter after filtration) and leukocyte count (39.9 +/- 11.3 x 10(6) vs. 1.3 +/- 0.9 x 10(6); p < 0.0005). Filtration caused a platelet loss of 16%, the platelet count decreasing not significantly from 2.91 +/- 0.75 x 10(11) to 2.40 +/- 0.94 x 10(11) (p = 0.26). After 5 days of storage all parameters of platelet function (platelet aggregation to several stimuli, hypotonic shock reaction [HSR] and platelet retraction), mean platelet volume, and pH and pCO2 showed no advantage for PCs filtered prior to storage compared to PCs stored unfiltered. Moreover, platelet aggregation on day 5 using 4 agonists at 10 concentrations showed worse results in 4 assays in prestorage filtered PCs (collagen [4 micrograms/ml: p < 0.05, ADP [0.2 mM]: p < 0.05, ADP [0.3 mM]: p < 0.05, thrombin [0.6 E/ml]: p < 0.05). But there is no convincing trend in all aggregation tests, and HSR, presumably the most useful parameter, was not different or day 5. CONCLUSIONS There is no advantage in terms of improved quality for prestorage leuko-depletion of PCs. Taking into account the obvious disadvantages of filtration, such as platelet loss and increasing costs per transfusion, we conclude that pre- or post-storage filtration of single-donor PCs should be done only for patients who have a clear indication for the transfusion of leukocyte-poor blood products.

One hundred and nine patients suffering from various malignancies underwent 285 apheresis procedures for PBPC collection. A median of two leukaphereses (range: 2-5) resulted in median numbers of 4.6 x 10(8) MNC/kg, 14.1 x 10(4) CFU-GM/kg, and 6.0 x 10(6) CD34+ cells/kg. Preleukapheresis peripheral blood CD34+ cells correlated significantly with collected CD34+ cells/kg (r = 0.94; p0.0001) and with CFU-GM/kg (r = 0.52; p0.0001). A value4 x 10(4) CD34+ cells/ml was highly predictive for a collection yield2.5 x 10(6) CD34+ cells/kg harvested by a single leukapheresis. Sixty patients were evaluated for hematologic reconstitution and engrafted in a median time of 10 days for WBC1.0 x 10(9)/l (range: 7-21 days), 10 days for ANC0.5 x 10(9)/l (7-20) and 11 days for PLT20 x 10(9)/l (7-62). Reinfused CD34+ cells/kg correlated significantly with hematologic engraftment (r = 0.44-0.52 and p0.006-0.001) as well as CFU-GM/kg (r = 0.36-0.44 and p0.007-0.001). A progenitor cell dose2.5 x 10(6) CD34+ cells/kg or8.0 x 10(4) CFU-GM/kg led to a significantly faster recovery for WBC, ANC, and PLT when compared with patients receiving2.5 x 10(6) CD34+ cells/kg or8.0 x 10(4) CFU-GM/kg. We conclude that rapid hematopoietic engraftment after high-dose therapy and PBPC reinfusion correlates well with a progenitor cell dose2.5 x 10(6) CD34+ cells/kg or8.0 x 10(4) CFU-GM/kg, and that above a preleukapheresis threshold of 4 x 10(4) CD34+ cells/ml a PBPC autograft containing2.5 x 10(6) CD34+ cells/kg can be collected by a single leukapheresis. We suggest that patients recovering from myelosuppression should be monitored for CD34+ cells in serial blood samples to determine the course of circulating hematopoietic progenitor cells. This issue will help to define the optimal time point to start apheresis and to predict a PBPC autograft harvested by a single leukapheresis, which will lead to rapid and stable hematopoietic reconstitution following transplantation.

PURPOSE To compare autologous bone marrow (BM) with peripheral-blood progenitor cells (PBPC) as hematopoietic rescue after high-dose chemotherapy (HDCT). PATIENTS AND METHODS From January 1991 until April 1993, 47 consecutive patients with relapsed or refractory germ cell tumors were randomized to either BM harvest or collection of PBPC mobilized by chemotherapy plus granulocyte colony-stimulating factor (G-CSF). After additional conventional-dose salvage treatment, all patients received HDCT with carboplatin 1,500 mg/m2, etoposide 2,400 mg/m2, and ifosfamide 10 g/m2 with either BM or PBPC rescue. RESULTS Forty-six patients were assessable for hematologic reconstitution, and one patient died on day +4 before engraftment. Rescue using PBPC resulted in a significantly shorter recovery time to neutrophil counts more than 500/microL (10.0 v 11.0 days, P < .01), neutrophil counts more than 1,000/microL (10.0 v 12.0 days, P = .001), and platelet counts more than 20,000/microL (10.0 v 17.0 days, P < .01), as well as in fewer days to transfusion independence from RBCs (8.0 v 12.0, P < .05) and platelets (9.0 v 12.0, P < .01) and fewer days of intravenous (IV) antibiotics (9.0 v 11.0, P < .05). However, no statistical differences in transfusion requirements or in other clinical outcome variables were observed. Overall survival and event-free survival also were not different in the two study arms. CONCLUSION We conclude that the use of PBPC mobilized by chemotherapy plus G-CSF results in sustained trilineage reconstitution after HDCT, which occurs more rapidly as compared with BM. The earlier hematologic reconstitution in patients with PBPC rescue significantly reduces the time to transfusion independence.

HIGH-DOSE CHEMOTHERAPY with stem cell support is a treatment option increasingly used for solid tumors in childhood and adolescence (1). However, after repeated cycles of myelosuppressive chemotherapy with or without radiotherapy, poor stem cell mobilization has been reported in up to 20% of patients (2,3). Risk factors have been identified as preceding cumulative high doses of alkylating agents and radiation therapy (4,5). In these cases, mobilization with granulocyte colony-stimulating factor (G-CSF) from steady state, mobilization following high-dose cyclophosphamide plus G-CSF, or a bone marrow harvest have been proposed with variable success. With regard to alternative stem cell mobilization protocols using new hematopoietic growth factors the combination of stem cell factor (SCF) plus G-CSF seems to have strong mobilization potential for first-line mobilization in patients with breast cancer, multiple myeloma and non-Hodgkin lymphoma (NHL) (6–10). It is unclear, however, whether SCF plus G-CSF would reliably mobilize stem cells in clinical situations where a conventional mobilization regime has already failed. Additionally, little information exists regarding the use, safety, and efficacy of the SCF plus G-CSF approach in a pediatric or adolescent setting. Here we report 3 patients with solid tumors who were successfully mobilized by SCF and G-CSF stimulation after failing mobilization with chemotherapy and G-CSF alone. The first patient, aged 17 years at initial diagnosis, suffered from relapsed stage IV B Hodgkin’s disease, which was treated initially with radiochemotherapy according to the GPOH-HD-95 protocol. Following relapse, she received additional IEP-ABVD-IEP-ABVD chemotherapy. Mobilization of CD34 progenitor cells with G-CSF after the second IEP chemotherapy cycle was not successful (Table 1). As an alternative, mobilization with SCF plus G-CSF was undertaken after the second ABVD cycle. Mobilization consisted of subcutaneous SCF (Ancestim, StemgenTM, Amgen Canada Inc., Mississauga, Ontario, Canada) (10 mg/kg s.c.) for 3 days alone, followed by 6 days of the combination of SCF (10 mg/kg s.c.) and GCSF (Filgrastim, Neupogen®, Amgen, Munich, Germany) (10–12 mg/kg s.c.) as described (7,8) under appropriate antiallergic premedication, as recommended by the drug manufacturer. Leukapheresis was performed on the 5th, 6th, and 7th day of the G-CSF stimulation with a total yield of 5.13 106 CD34pos cells/kg b.w. At this time, the cumulative chemotherapy dose had reached 9 g/m2 procarbazin, 21 mg/m2 vincristin, 260 mg/m2 adriamycin, 4 g/m2 cyclophosphamide, 20 g/m2 ifosfamide, 1.25 g/m2 etoposide, 40 mg/m2 bleomycin, 24 mg/m2 vinblastin, and 1.5 g/m2 dacarbazin. The second patient, aged 14 years at diagnosis, suffered from relapsed Ewing’s sarcoma of the right humerus and the brain. Firstline therapy consisted of 14 EVAIA cycles plus local radiation with 44.8 Gy according to the EICESS 92 protocol; second-line treatment was 4 Cyc-ETO cycles

The objective was to investigate potential risks for apheresis donors associated with a triple-plateletpheresis (TP) program.Eleven hemapheresis centers randomly assigned 411 repeat donors (ratio, 1:1.2) to either double plateletpheresis (DP; 185 donors) or TP (226 donors) with a platelet (PLT) target content of at least 5.0×10(11) PLTs/DP and at least 7.5×10(11) PLTs/TP. The primary endpoint was procedure-related postapheresis PLT count of at least 150×10(9) /L (probability, ≥98%). Secondary endpoints were apheresis characteristics and donor adverse reactions.In 6 of 1133 DPs (0.5%) in 4 of 185 donors (2.2%) and in 20 of 1020 TPs (2.0%) in 14 of 226 donors (6.2%), postapheresis PLT counts were below 150×10(9) /L. There were marginal but significant differences in collection efficiency (DP, 69.2±9.1%; TP, 70.9±9.0%; p≤0.0001) and collection rate (DP, 10.4×10(9) ±2.3×10(9) PLTs/min; TP, 10.8×10(9) ±2.3×10(9) PLTs/min; p≤0.005). The PLT yields were 5.9×10(11) ±0.8×10(11) PLTs for DP and 8.3×10(11) ±0.9×10(11) PLT for TP (p≤0.0001) at processing times of 59±13 minutes (DP) versus 80±16 minutes (TP; p≤0.0001). Significant PLT recruitment (1.10±0.14 vs. 1.20±0.23; p0.0001) was seen for both DP and TP. DP and TP did not differ with regard to venous access problems (VAPs) without discontinuation (3.8% for both), but DP induced fewer VAPs with discontinuation (1.1% vs. 3.0%; p0.01). Mild citrate toxicity (1.7% vs. 3.9%; p0.01) and circulatory reactions (0.4% vs. 2.2%; p0.01) were more often noticed in TP, but caused no increase in discontinuations.TP results in an increase in mild donor reactions but does not significantly impair donor safety or product quality.

In the context of a controlled multicenter study on intravenous immunoglobulin (IVIG) treatment of patients with a history of unexplained recurrent spontaneous abortions (RSA), a number of controversial immunological parameters were evaluated prior to and during pregnancy with respect to their diagnostic and/or prognostic significance. A total of 390 serum samples from 52 patients were investigated. Sharing of 2 or more HLA (A, B, DR, DQ) antigens was significantly more frequent in RSA couples than in controls. The rate of cytotoxic or Fc-receptor (FcR)-blocking antibodies was not significantly lower in RSA patients than in individuals with normal pregnancies. Both tumor necrosis factor-alpha (TNF-alpha) levels and IgG anticardiolipin antibodies (IgG-ACA) were significantly increased in the patient group. While the occurrence of HLA sharing, cytotoxic/FcR-blocking antibodies and IgG-ACA did not correlate with the outcome of pregnancy, TNF-alpha levels were found to be significantly higher in patients with subsequent miscarriage than in those with successful pregnancy. IgG-ACA, if present, significantly decreased during the course of successful pregnancy but remained high in patients with subsequent abortion. It is concluded that the diagnostic and/or prognostic value of HLA sharing and cytotoxic/FcR-blocking antibodies has been overestimated while TNF-alpha and ACA levels are potential diagnostic markers and/or exhibit prognostic significance in subgroups of RSA patients.

Background: Antibodies to HLA determinants may decrease the increment after platelet transfusion and are correlated with increased rates of rejection in renal transplantation. Cross-match tests and HLA antibody specification are used to identify compatible donors for sensitized patients. However, search for cross-match negative donors may be ineffective, and many sera containing antibodies toward public HLA epitopes give no clear results in conventional specificity analysis. Materials and Methods: We tested a series of 4,625 sera from 1,073 patients with hematological, malignant or other diseases using a fluorescence lymphocytotoxicity test (LCT). We applied an evaluation program incorporating a list of public antigens belonging to known cross-reacting groups (CREGs) to detect antibodies toward private and just as well public epitopes. Results: In 694 sera (15.0%) from 240 patients the panel reactivity (PRA) in LCT assay was higher than 5%. PRA was > 50% in 258 (37.2%) and < 50% in 436 sera (62.8%). In both groups we identified specific antibodies toward public HLA class I epitopes. Overall antibody specification was successful in 429 of 694 sera (61.8%) and in 175 of 240 patients (72.9%), respectively. The rate of antibodies against public epitopes shared by more than one HLA class I gene product was 203/694 (29.3%) with respect to tested sera and 83/240 (34.5%) with respect to patients. The rate of public epitope antibodies was highest in sera with PRA values from 30 to 90% showing public epitope specificity in 159 of 353 sera (45.0%). Conclusions: We conclude that antibodies toward public HLA class I determinants are detectable not only in highly sensitized patients. The described program may increase the rate of antibody specification and facilitate the platelet supply in platelet refractory patients.

Hintergrund: Seit Oktober 1991 steht für den Zellseparator Fresenius AS 104 ein Single-needle-(SN-)Programm zur Verfügung. Wir berichten über unsere Erfahrungen während der ersten 18 Monate im Routinebetrieb mit dem SN-Verfahren. Material und Methodik: Im Zeitraum von 18 Monaten wurden bei 225 Blutspendern 395 Thrombozytapheresen im SN-Verfahren durchgeführt. Der Einfluß unterschiedlicher Flußgeschwindigkeiten, Zyklusvolumina und Trenngrenzenpositionen auf die Effizienz der Thrombozytapherese und auf die leukozytäre Kontamination der Konzentrate wurde untersucht. Ergebnisse: Bei 395 Thrombozytapheresen konnten im Mittel 3,00 ± 0,69 ×1011 Thrombozyten bei einem durchschnittlichen Konzentratvolumen von 283 ± 39 ml und einer Leukozytenkontamination von 1,66 ± 3,1 ×108 Leukozyten pro Konzentrat gewonnen werden. Flußgeschwindigkeiten von 50 ml/min (versus 60 ml/min) sowie eine Trenngrenzenposition von 6:2 (versus 7:1) erbrachten verminderte leukozytäre Kontaminationen der Konzentrate, ohne jedoch statistische Signifikanz zu erreichen. Schlußfolgerung: Das SN-Programm des Zellseparators AS 104 zeichnet sich durch eine hohe Thrombozytenausbeute aus und zeigt im Vergleich zu anderen SN-Systemen geringere Leukozytenwerte im Konzentrat. Eine weitere Verminderung der Leukozyten im Konzentrat ist jedoch anzustreben, da 77% der Konzentrate den empfohlenen Grenzwert von 0,5×108 Leukozyten pro Konzentrat überschreiten.

We have determined the incidence and clinical significance of positive microbiologic cultures in a series of 290 peripheral blood stem cell concentrates in 95 patients undergoing multiple apheresis procedures for autologous stem cell rescue. Specimens for bacterial cultures were obtained after processing of the autografts just prior to freezing. The incidence of microbial contamination was 4.5% (n = 13). The predominant pathogenic microorganism cultured was coagulase-negative Staphylococcus (n = 11). From 8 patients with contaminated leukapheresis products 6 underwent autologous stem cell transplantation. Five patients received 1-5 culture-positive stem cell concentrates without serious sequelae, whereas the sixth patient was autografted with noncontaminated leukapheresis products, 1 concentrate contaminated with Aspergillus fumigatus being not reinfused. No microorganism present in the stem cell autograft was recovered in vivo in the posttransplantation period, although fever as a sign of infection occurred in all but 1 patient. Peripheral blood stem cell collection and ex vivo processing for cryopreservation may result in microbiologic contamination. However, our data show that infusion of contaminated stem cell autografts does not play a significant role as a source for infections in the clinical setting of autologous stem cell rescue.

Antibodies against the human neutrophil antigens (HNA) are able to stimulate transfusion reactions, autoimmune and neonatal neutropenia. The aim of this study was to determine the HNA allele frequencies in the largest ethnic minority group in Germany in comparison with the German population for predicting the risk of alloimmunization and associated transfusion reactions, as well as the risk of developing neonatal neutropenia for the newborn of racial mixed couples. However, there exists no data about HNA genotype distribution in Turkish population. DNA was isolated from blood samples of 119 German and 118 Turkish blood donors and typed them for HNA-1, -3, -4, and -5 by using a commercial polymerase chain reaction kit with sequence-specific primers (SSP-PCR) and compared the HNA genotype distribution of both groups. In German blood donors, the gene frequencies for HNA-1a and HNA-1b were 0.391 and 0.601, for HNA-3a and -3b, 0.744 and 0.256, for HNA-4a and -4b, 0.908 and 0.092, and for HNA-5a and -5bw, 0.731 and 0.269. In Turkish blood donors, we observed 0.420/0.564, 0.737/0.263, 0.881/0.119, and 0.754/0.246 for HNA-1a/1b, -3a/3b, -4a/4b, and -5a/5bw. No statistic significant difference between genotypes in these populations was observed. This study is the first to report HNA gene frequencies in a Turkish population. It showed that there is no difference of HNA genotype in blood donors with Turkish descent in comparison with German blood donors. The alternating transfusion of blood and blood components is no increased risk for developing alloantibodies against HNA antigens. In pregnancy of mixed couples no special screening programs for HNA are necessary.

But de l'etude. - Le syndrome d'hyperstimulation ovarienne est une complication, potentiellement mortelle, qui peut se produire au cours de la stimulation ovarienne, pratiquee dans le cadre d'un traitement de l'infertilite. Puisqu'aucune association entre ce fait et les groupes sanguins du systeme ABO n'etait connue, nous avons compare les antigenes ABO avec l'importance et l'apparition des symptomes, dans une etude de cas. Patients et methodes, - Entre janvier 2000 et fevrier 2007, 121 patientes, principalement caucasiennes, ont ete hospitalisees en raison du syndrome d'hyperstimulation ovarienne apres fecondation in vitro. La severite des symptomes, les taux de grossesse et les groupes sanguins ABO ont ete classes. Le groupe sanguin ABO a ete compare a quatre groupes de temoin independants. Resultats. - Chez les malades souffrant du syndrome d'hyperstimulation ovarienne, on retrouve plus frequemment le groupe sanguine A, plutot que le groupe 0. Pour les patientes du groupe A qui subissent une stimulation ovarienne controlee, le risque de developper un syndrome d'hyperstimulation d'apparition recente est augmente par rapport aux patientes avec le groupe 0 (odds ratio 2,171, valeur p = 0,002). Aucune association n'a ete trouvee concernant une apparition tardive. Dans le groupe d'apparition tardive du syndrome d'hyperstimulation ovarienne, le taux global de grossesse est de 50,4 %, donc trois fois superieur au groupe d'apparition recente. Quatre patientes, aucune d'elles n'avait de groupe sanguin 0, ont developpe une thrombose de la veine jugulaire ou subclaviere. Conclusion. - Le groupe sanguin A peut etre associe au syndrome d'hyperstimulation ovarien d'apparition recente parmi les Caucasiens. En fonction des resultats d'etudes a venir, une association possible peut etre consideree, voire une posologie en hormone individualisee pour la stimulation ovarienne controlee.

The aim of this study was to investigate the effect of gamma irradiation with 30 Gy on the coagulation system in leukoreduced fresh-frozen plasma (FFP).In 74 FFP units that had been stored for 352 +/- 103 days below -30 degrees C, the following variables were determined in parallel in an irradiated and not irradiated half: prothrombin time (PT); activated partial thromboplastin time (APTT); thrombin time; antithrombin III; protein C; protein S; von Willebrand factor antigen; ristocetin cofactor; plasminogen-alpha(2)-antiplasmin; the coagulation factors fibrinogen, factor (F)II, FV, FVII, VIII, F IX, FX, FXI, FXII, FXIII, and activated factor XII (FXIIa); D-dimer; fibrin monomer; thrombin-antithrombin complex; prothrombin fragment 1 + 2 (F1+2); plasmin-alpha(2)-antiplasmin complexes (PAPs); and platelet factor 4. The FVII activity ratio was assayed to quantify activation of FVII.Irradiation with 30 Gy resulted in a reduction of APTT (35.0 +/- 4.1 sec vs. 34.4 +/- 4.1 sec; p = 0.00000006) and PT (89.8 +/- 8.2% vs. 90.7 +/- 8.0%; p = 0.002) and a significant increase of the activities of the coagulation factors FII, FV, FVII, F IX, FX, and FXII. FVIII activity decreased from 118 +/- 31 to 116 +/- 32 percent (p = 0.02). Activation of the coagulation system was shown by an increase in the FVII activity ratio (1.19 +/- 0.29 vs. 1.31 +/- 0.34; p = 0.0000001), FXIIa (0.81 +/- 0.50 ng/mL vs. 0.90 +/- 0.51 ng/mL; p = 0.006), and F1+2 (1.19 +/- 0.20 nmol/L vs. 1.24 +/- 0.20 nmol/L; p = 0.000005) after irradiation with 30 Gy, whereas an increase of PAP (16.2 +/- 11.5 ng/mL vs. 20.2 +/- 12.0 ng/mL; p = 0.0004) demonstrated activation of the fibrinolytic system. No negative influence of irradiation with 30 Gy on inhibitors of coagulation was observed.Gamma irradiation of leukoreduced FFPs with 30 Gy results in a significant but very weak activation of the coagulation and fibrinolytic system in FFPs.

BACKGROUND: This prospective study compared white blood cell (WBC) storage in polyvinylchloride (PVC) bags and in polyolefin (POF) bags. After leukapheresis, CD14+ monocytes, CD11c+, and CD123+ precusor dendritic cells (DCs) were analyzed under platelet (PLT) storage conditions. STUDY DESIGN AND METHODS: Twenty-five leukapheresis procedures were performed on blood cell separators (AS.TEC204 [PVC; Fresenius HemoCare GmbH] and the COBE Spectra [POF, Gambro BCT]). Blood cell counts, glucose, lactic acid, pO2, pCO2, and pH were measured in mononuclear cell (MNC) harvests on Days 0, 1, 3, and 5. WBCs were analyzed by flow cytometry. RESULTS: The WBC yields of the AS.TEC204 harvests were 25 percent higher (13.4 × 109 ± 2.7 × 109 WBCs) compared to the COBE Spectra (9.9 × 109 ± 2.5 × 109 WBCs). During 5-day storage, WBC counts (PVC bags) decreased significantly (24%). Storage in POF bags showed more consistent results (WBC loss, 6%). Loss of CD14+ monocytes and CD11c+ precursor DCs did not differ significantly in leukapheresis products. CD123+ precursor DCs stored in PVC bags dropped by more than 90 percent (POF bags, 24%). Lactic acid concentrations exceeded 20 mmol per L after 24 hours in PVC bags and after 72 hours in POF bags. The mean pH value on Day 5 was 6.2 (PVC) and 6.3 (POF). On Day 1, the product glucose concentration decreased by 76 percent after storage in PVC bags and by 16 percent in POF bags. CONCLUSIONS: Storage of MNCs within 72 hours in the original harvest container assures stable WBC content and is easy to perform. POF bags should be preferred in the case of extended WBC storage. Patient studies should clarify changes in efficiency of hematopoietic reconstitution that might occur over time during MNC storage.

SUMMARY. To compare the performance of seven currently available test systems in the detection of erythrocyte alloantibodies (ab), we tested in parallel 446 sera samples containing red cell ab [368 sera samples with ab that are assumed to be clinically significant (cs-ab) and 78 sera samples with ab that are assumed to be of minor clinical significance (ms-ab)] using the tube spin low-ionic-strength solution (addition method) indirect antiglobulin test (tube LISS-IAT), three microtube column agglutination techniques (DiaMed-ID, Ortho BioVue and Bio-Rad Scangel), one affinity adherence test system (CLB/Mast CellBind Screen) and two solid-phase tests [Biotest Solidscreen II and Immucor Capture-R Ready-Screen (4)]. To address the specificity of the three test systems under routine conditions, results of 4566 patient samples obtained using the tube LISS-IAT, results of 5205 patient samples obtained using the Scangel and results of 3560 samples obtained using the Capture-R were evaluated. The DiaMed-ID detected 344 cs-ab and 43 ms-ab, BioVue 333 cs-ab and 48 ms-ab, Scangel 348 cs-ab and 62 ms-ab, CellBind Screen 346 cs-ab and 47 ms-ab, Solidscreen 330 cs-ab and 38 ms-ab, Capture-R 358 cs-ab and 45 ms-ab and LISS-IAT 159 cs-ab and 12 ms-ab. In routine practice, erythrocyte cs-ab could be identified in 61 (67� 8%) of 90 reactive sera (specificity: 98� 6%) in the tube LISSIAT, in 169 (58� 7%) of 288 (94� 4%) in Bio-Rad Scangel and in 101 (51� 0%) of 198 reactive sera (94� 3%) in Capture-R. We conclude that the sensitivity of the microcolumn, affinity adherence and solid-phase test systems in the detection of cs-ab was similar and was markedly superior to that of the conventional tube LISS-IAT. All high-sensitive test systems produced higher rates of false positives and ms-ab compared to the tube test. An individual cost–benefit analysis, considering the recent knowledge about the clinical significance of weak-reactive cs-ab, should be performed in every institution to decide whether and if so which high-sensitive screening system should be applied.

Rare clinical conditions cause the need for washed platelet (PLT) concentrates (PCs). Saline-washed PCs can only be stored shortly, however, owing to lack of substrates for PLT metabolism. New PLT additive solutions (PASs) contain such substrates and might be used alternatively. The in vitro quality of apheresis PCs washed with Composol-PS or modified PAS-III (PAS-IIIM) stored up to 48 hours after wash was compared.Twelve blood donors underwent two apheresis procedures (A and B) collecting 6.0 x 10(11) PLTs in 500 mL of plasma with a least 2 weeks in between. The PCs collected by Apheresis A were stored for 3 days and then split in two equal units before washing with Composol-PS or PAS-IIIM. The PCs collected by Apheresis B were split after collection. One unit was released for transfusion and 1 unit was stored unwashed up to Day 6 and used as reference unit. In vitro testing was performed before and after washing as well as 24 and 48 hours after wash.After 48 hours of postwash storage, the units washed with either PAS showed acceptable results for hypotonic shock response (HSR), P-selectin expression, and pH, whereas PLT aggregability was significantly impaired. Throughout the storage, unwashed units showed better in vitro quality. HSR and P-selectin expression were similar before and immediately after the washing procedure.Based on these in vitro results, 48-hour postwash storage of washed PCs with the two PASs seems to be feasible. In vivo recovery studies, however, must confirm this finding in the future.

The COM.TEC cell separator (Fresenius Hemocare), equipped with the MNC program (single-stage chamber) and the PBSC-LYM program (dual-stage chamber), was evaluated for CD 14+ cell collection regarding cell yields, collection efficiencies (CEs), and the content of residual cells in harvests.Twenty-four non-cytokine-stimulated donors underwent 5-L mononuclear cell (MNC) collections on the COM.TEC device to compare both programs. Two software versions (v02.03.05 vs. v 03.00.04) were investigated for optimization of the CD 14+ cell collection process. Blood counts of donors and products were analyzed for CD 14+ cells by flow cytometry and for platelets (PLTs), white blood cells, and red blood cells (RBCs) by a blood cell counter.In 5-L collections, the MNC program resulted in high CEs (83+/- 23%) and yields (1.2 x 10(9)+/- 0.6 x 10(9) per unit) of CD 14+ cells, but the products showed high residual PLTs. The use of a dual-stage chamber in the PBSC-LYM program produced a low content of residual PLTs (0.7 x 10(11) +/- 0.3 x 10(11) per unit) and RBCs but failed to reach a target of 1 x 10(9) CD 14+ cells. Modulated light to stabilize the buffy-coat detection by the interface monitor significantly improved CD 14+ cell enrichment. By use of the PBSC-LYM program, higher centrifuge velocity (1700 rpm [382 x g] vs. 1500 rpm [297 x g]) improved significantly CD 14+ cell yields (0.7 x 10(9) vs. 0.5 x 10(9) cells).A pure CD 14+ cell product with low numbers of residual cells was obtained by the PBSC-LYM program, which could be useful for good manufacturing practice-conformed production within closed systems. The MNC program offers the collection of high CD 14+ cell yields with excellent CEs but also high residual PLT counts.

BACKGROUND AND OBJECTIVES: New platelet (PLT) additive solutions (PASs) allow a plasma carryover of < 30% in PLT concentrates. This implicates the need to collect apheresis PLT concentrates at very high PLT concentrations: so-called dry PLTs (DPs). We used the TRIMA, with software version 4 (TRIMA V4), to collect such DPs and investigated the in vitro quality of these PLTs when stored in the new modified PAS-III (PAS-IIIM). MATERIALS AND METHODS: TRIMA V4 was programmed to collect 6.0 x 10(11) PLTs at a concentration of 5000 x 10(3) PLTs/microl. Two DPs were pooled, split into four equal parts and diluted to obtain secondary pools (SPs) consisting of 70% PAS-III/30% plasma, 70% PAS-IIIM/30% plasma, 80% PAS-IIIM/20% plasma or 100% plasma. In vitro testing was performed on days 0, 1, 5 and 7. Collection efficiency (CE), collection rate (CR) and PLT yield were calculated for each donation. RESULTS: Thirty-two runs with TRIMA V4 were performed, collecting 6.58 +/- 0.74 x 10(11) PLTs at a concentration of 4255 +/- 914 x 10(3)/microl in 99 +/- 19.9 min, resulting in a CE of 65.3 +/- 8.2% and a CR of 6.92 +/- 1.6 x 10(9) PLTs/min. On day 0, 34-37% of the PLTs in the units prepared for storage were already activated. PLTs stored in 70% or 80% PAS-IIIM showed superior in vitro quality compared to PLTs stored in PAS-III. CONCLUSIONS: TRIMA V4 is a suitable device for the collection of DPs. Nevertheless, improvements are desirable to further increase the ability to concentrate PLTs at very high levels. The storage of apheresis-derived PLTs in PAS III-M is a very promising approach, even at a plasma carryover of < 30%.

BACKGROUND: We introduced monitoring of mononuclear cell (MNC) counts to obtain enhanced donor control and a stable quality of MNC products, because there are limited data available about blood donors after serial leukapheresis (LP) procedures. STUDY DESIGN AND METHODS: In a prospective paired study, 13 male healthy blood donors underwent 10-L LP procedures performed on two apheresis devices by use of two MNC program settings (COBE Spectra, Gambro BCT, SF 250 vs. SF 500; and AS.TEC 204, Fresenius Hemocare, CP 129 vs. CP 194). Donors’ pre- and postdonation MNC counts were analyzed by fluorescence-activated cell sorting. RESULTS: After each 10-L LP procedure, a transient decline (p

In cancer and transplantation therapy apheresis devices and software of optimum standards are required for the collection of high cell yields with high purity of the desired cell fraction.In a paired study, 15 healthy blood donors underwent four 10-L leukapheresis procedures (197 +/- 33 min) with an inlet blood flow rate of 60 mL per minute by use of two different MNC program settings of the COBE Spectra (Gambro BCT) and the AS.TEC 204 (Fresenius Hemocare) cell separators.Use of the standard MNC program of both apheresis devices resulted in significantly higher (p0.01) collection efficiencies of CD14+ monocytes, CD3+ cells, CD4+ cells, CD8+ T cells, CD16+ CD56+ natural killer (NK) cells, and residual PLTs (p0.001), owing to higher centrifuge speed. The mean MNC purity of all components was more than 90 percent. By use of standard programs of either device, significant correlations (p0.01) between donor monocytes and preleukapheresis NK cell counts and the corresponding component cell yields were found.Compared to the program modifications with lower centrifuge velocities the standard MNC programs were significantly more efficient regarding CD14+, CD3+, and CD16+ CD56+ cells. Enhanced centrifuge speed and inlet blood flow rate in MNC programs resulted in higher, similar composed MNC concentrations of the products.