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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

Background: Sensitive and accurate methods to detect hematopoietic chimerism after hematopoietic stem cell transplantation (HSCT) are essential to evaluate engraftment and to monitor response to therapeutic procedures such as donor lymphocyte infusion. Continuous long-term follow up, however, requires large amounts of pre-HSCT samples limiting the application of many widely used techniques for sensitive chimerism monitoring. Methods: DNAs from 42 normal healthy donors and 16 HSCT donor/recipient pairs were employed to validate the use of allele-specific insertion/deletion (indel) quantitative real-time polymerase chain reaction (qPCR) to quantify chimerism in samples with low amounts of DNA. Consequently, indel-qPCR analyses of samples from 16 HSCT patients were compared to short-tandem repeat (STR) specific PCR analyses. Results: Typing with reduced amounts of input DNA (15 vs. 60 ng) allowed for the reliable distinction of positive (mean threshold cycle (ct) 28.05) and negative (ct >36) signals. The high informativity of primer/probe sets, with 12 out of 19 markers exceeding 20% informativity, was confirmed in our cohort (n = 74). Importantly, a fourfold reduction of input DNA compared to published protocols did not alter PCR efficiencies and allowed for a more sensitive detection of chimerism in 7 of 16 HSCT patients compared to results obtained by STR-PCR. Conclusions: Our data suggest that indel-qPCR is a more sensitive technique for the detection of hematopoietic chimerism compared to STR-PCR and works efficiently for samples with low amounts of DNA.

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: 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: 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: 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 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 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.

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.

BACKGROUND: There exists only very few data on in vitro and in vivo effects of gamma irradiation of red blood cells (RBCs) that have been leukoreduced by filtration before a subsequent irradiation. Reported studies reflect neither the current Food and Drug Administration (FDA) nor the European recommendations on timing of irradiation and subsequent storage. STUDY DESIGN AND METHODS: We studied 40 RBC units that were prepared from inline filtered whole blood and 40 RBC units that were filtered after component separation. All RBCs were stored in the additive solution saline-adenine-glucose-mannitol and leukoreduced on the collection day. In both groups, 20 components were irradiated on Day +14 with 30 Gy, and 20 served as nonirradiated controls. In vitro evaluation of both irradiated and nonirradiated RBC units was performed before and after irradiation on Days +1, +7, +14, +21, +28, +35, and +42 from the collection day. RESULTS: Gamma irradiation induced enhanced leakage of potassium ions and lactate dehydrogenase and an enhanced in vitro hemolysis rate in the irradiated components. However, in vitro hemolysis rate of both nonirradiated and irradiated components was remarkably lower than 0.8 percent, and the preservation of adenosine triphosphate over 42 days was satisfying. CONCLUSIONS: This study reflects the current FDA and European recommendations on timing of irradiation and subsequent storage. Our findings together with recent results of other investigations on the effect of gamma irradiation on leukoreduced RBCs allow the proposal that a storage time up to 28 days after irradiation is allowable.

BACKGROUND: Semaphorin 7A (Sema7A), the protein that carries the JMH blood group antigen, is involved in immune responses and plays an important role in axon growth and guidance. Because previous serologic studies on red blood cells (RBCs) suggested a considerable diversity of Sema7A, the present study was designed to elucidate the complex picture of the molecular diversity of this protein. STUDY DESIGN AND METHODS: The JMH antigen status was determined by serology, flow cytometry, and Western blot. Genomic and transcript analysis of SEMA7A was performed by nucleotide sequencing. Recombinant Sema7A proteins were used for genotype-phenotype correlation. A three-dimensional model of Sema7A was generated for topologic analyses. RESULTS: Our studies on 44 individuals with unusual JMH phenotypes and their family members revealed that aberrant Sema7A expression can be an inherited or an acquired phenomenon and is based on reduced surface expression or qualitative changes in Sema7A. These different phenotypes are caused by variations of the SEMA7A gene or seem to be generated by autoimmune-related or RBC lineage-specific mechanisms. The variant JMH phenotypes were related to the presence of missense mutations in SEMA7A, predicting amino acid changes in the semaphorin domain of Sema7A. Sequence analysis of the variant SEMA7A alleles revealed mutations affecting codons 207 and 460/461. Topologic analyses showed that Sema7A polymorphisms were prominently located on the top and bottom of the semaphorin domain, suggesting a functional relevance of these sites. CONCLUSION: These findings provide a basis with which to delineate the various ligand-binding surfaces of Sema7A.

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: 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

BACKGROUND: Recently, long-standing elevations of soluble growth factors released from platelets (PLTs) after contact with artificial surfaces during dialysis were described. They could be jointly responsible for the high frequency of death from cardiovascular diseases in dialysis patients. There are no comparable data on the extent and the duration of a growth factor release by plateletpheresis procedures. STUDY DESIGN AND METHODS: A total of 37 plateletpheresis procedures were performed with two different devices. PLT-derived growth factor (PDGF) isoform AB, transforming growth factor (TGF)-beta1, and beta-thromboglobulin (beta-TG) were measured in the donors' plasma samples, and PLT activation and function were measured by cytometry and aggregometry before and after plateletpheresis and 1 and 24 hours later. RESULTS: Before apheresis, the following mean plasma levels were found: beta-TG, 98.6 +/- 37.3 IU per mL; PDGF-AB, 71.5 +/- 38.5 pg per mL; and TGF-beta1, 2.24 +/- 0.80 ng per mL. At the end of the apheresis procedures, the mean PDGF-AB level had increased by a factor of 1.8 (p < 0.05). One hour later, the mean PDGF-AB level had normalized again. No significant change in the levels of beta-TG and TGF-beta1 was found by the apheresis procedures. There was no influence of the blood cell separator type on the results. CONCLUSION: Only a slight and rapidly reversible increase in soluble PDGF-AB was found during plateletpheresis and no increase in soluble TGF-beta1 and beta-TG was found. This change should not be harmful to the donor.

We report the case of a 53-year-old patient suffering from severe hemophilia A showing six different blood group antibodies (anti-c, -Cw, -Jkb, -E, -Lea -Kpa). The decision to replace his left kneeendoprosthesis was made in September 1993. Having multiple erythrocyte antibodies he cannot be transfused with allogenic blood; on the other hand, a large intraoperative blood loss could be expected due to severe hemophilia. An intensified regimen for preoperative autologous blood donation including oral and intravenous iron supplementation, application of recombinant human erythropoietin, and cryopreservation of packed red cells was performed. The patient reached a predeposit of 23 units of autologous packed red cells and 20 units of autologous fresh frozen plasma. Surgery took place in February 1994. Estimated peri-operative blood loss was about 11 liters; the patient received autologous blood exclusively: 13 units of packed red cells and 20 units of fresh frozen plasma. Acute intraoperative hemodilution was performed yielding 2×500 ml of autologous blood; 2,200 ml of autologous packed red cells could be retransfused after intraoperative salvage. This case report shows the enormous possibilities of an intensified regimen in autologous blood donation using recombinant erythropoietin and intravenous iron supplementation.

BACKGROUND: Umbilical cord blood processing with standard centrifugation techniques is performed in open systems and results in varying cell and volume recoveries. STUDY DESIGN AND METHODS: Forty umbilical cord blood donations were randomly assigned to processing either with a microprocessor-controlled cell separator equipped with closed disposables or with a manual separation procedure in blood bags. The collection efficiency of nucleated cells, MNCs, RBCs, and CD34+ cells and the processing time were analyzed. RESULTS: Using the cell processor, mean collection efficiencies were 78.6 ± 24.9 percent for nucleated cells, 77.4 ± 27.8 percent for MNCs, 55.5 ± 14.6 percent for RBCs, and 83.6 ± 32.5 percent for CD34+ cells, while they were 73.1 ± 13.2 percent for nucleated cells, 78.1 ± 14.9 percent for MNCs, 26.0 ± 12.2 percent for RBCs, and 77.0 ± 17.6 percent for CD34+ cells when using the standard centrifugation technique. The processing time was about 20 minutes for automated processing and 60 to 80 minutes for the standard centrifugation technique. CONCLUSION: Using the new cell processor, the collection efficiencies for nucleated cells, MNCs, and CD34+ cells are similar to those obtained by established centrifugation techniques while the RBC reduction is less effective. The main advantages of the new systems are the closed system, the more standardized processing procedure, and a significantly shorter processing time.

BACKGROUND To reduce the production costs of single-donor platelets (SDPs), a study was conducted to investigate whether plasma collected as a by-product of plateletpheresis satisfies the quality requirements for FFP without impairing the quality of the SDP component. STUDY DESIGN AND METHODS Ninety-two donors with platelet (PLT) counts

Background The effect of gamma radiation on single-donor apheresis platelet concentrates (SDPs) has been elucidated only incompletely. The only existing report on the function of SDPs stored in the irradiated state found a deterioration in the in vitro aggregability at the end of shelf life in SDPs divided before irradiation with 1500 cGy. Study design and methods The in vitro properties of platelets were examined in four series of irradiated and control platelets, each obtained from the same 15 donors. Irradiation with 3000 cGy was performed on Days 0, 3, and 5. Cellular content, aggregability by ADP alone or ADP and epinephrine, spontaneous and induced CD62 expression, beta-thromboglobulin release, glucose consumption, lactate production, and pH were measured immediately after preparation and on Days 3 and 5 after donation. Results Comparable in vitro properties were measured in irradiated and control platelets, whether irradiation was performed on Day 3 or Day 5. However, in platelets irradiated on Day 0, we found a significantly better in vitro aggregability by 20 microM: ADP immediately after irradiation and by 10 microM: ADP and 2 microM: epinephrine at the end of shelf life than was found in the other groups (Day 5 results: Day 0 irradiation: 75 +/- 32%; Day 3 irradiation: 45 +/- 45%; Day 5 irradiation: 47 +/- 41%; control: 40 +/- 24%; p Conclusion Gamma radiation had no adverse effect on platelet quality in extremely WBC-reduced SDPs. On the contrary, a slight, but significantly better in vitro aggregability was found in SDPs irradiated before storage than in platelets irradiated later during storage and in unirradiated platelets. This increased in vitro aggregability persisted until the end of shelf life.

BACKGROUND MNCs for adoptive immunotherapy may be collected by leukocytapheresis with a cell separator. STUDY DESIGN AND METHODS Six healthy cytapheresis donors donated two MNC concentrates on a cell separator (AS.TEC 204, Fresenius): one on the standard MNC program and one on a modified MNC program with reduced centrifuge velocity that leads to a lower platelet contamination. Seventeen patients with malignant melanoma donated 26 MNC concentrates: 5 on the AS.TEC 204 MNC program, 9 on the modified AS.TEC 204 MNC program, and 12 on another modified MNC program (Spectra, COBE). RESULTS In the course of cultivation of MNCs to dendritic cells (DCs), the donor MNC concentrates with the lower platelet contamination (475 +/- 85 x 10(9)/L) had a significantly higher relative DC yield (low platelet contamination: 3.9 +/- 1.6% of the plated cells; high platelet contamination: 2.5 +/- 1.8% of the plated cells; p = 0.019) than the concentrates with the higher platelet contamination (2364 +/- 448 x 10(9)/L). No significant difference was found in the yields of MNCs and CD14+ cells in the three protocols used for the collection of MNCs from patients with melanoma. The components obtained by the standard AS.TEC 204 MNC program had a significantly higher platelet contamination (1768 +/- 994 x 10(9)/L) than the components obtained by the modified AS.TEC MNC program (360 +/- 98 x 10(9)/L; p

Wir untersuchten Thrombozytenverlust, Leukozytendepletion, Thrombozytenmorphologie und -funktion (Aggregation, hypotone Schockreaktion, Retention und Retraktion) an 8 Paaren von Einzelspenderthrombozytapheresekonzentraten (Cobe Spectra, Fresenius AS 104) vor und nach Filtration durch zwei verschiedene Polyesterfilter (Sepacell PL-5A, Pall PL100). Ebenso verglichen wir den Transfusions-erfolg der unterschiedlich gefilterten Konzentrate anhand des Plättcheninkrementes 1 h nach Transfusion. Die Leukozytendepletion war bei akzeptablen Thrombozytenverlusten bei beiden Filtern effektiv ( > 99,9%), keines der eingesetzten Testsysteme erbrachte einen Hinweis auf eine Beeinträchtigung der Thrombozytenfunktion durch die Filtration.

Autologe Blutspende vor elektiven chirurgischen Eingriffen ist heute transfusions-medizinischer Standard. Während nur 6% aller elektiv operierten orthopädischen Patienten nicht in der Lage sind, 3 Eigenblutkonserven zu spenden, sind 4 Konserven für 40% dieser Patienten aufgrund spendebedingter Anämisierung nicht erreichbar [1, 2]. Entsprechend unseren Ergebnissen liegen dem physiologische Probleme der Dynamik der hämatopoetischen Regeneration zugrunde, die nicht allein durch unselektierten Einsatz humanen rekombinaten Erythropoietins zu bewältigen sind.

BACKGROUND: The purpose of this study was to compare the performance of four currently available microtube column agglutination systems in the detection of red cell alloantibodies to that of the standard tube low-ionic-strength solution (LISS) indirect antiglobulin test (IAT) (tube LISS-IAT). STUDY DESIGN AND METHODS: In a comparative study, 172 sera, previously demonstrated to contain red cell alloantibodies, were tested in parallel by the tube LISS-IAT and three microtube column agglutination techniques (DiaMed-ID, Ortho BioVue, and Sanofi-Pasteur Scangel) and one affinity-adherence test system (Gamma ReACT). Tests were performed simultaneously by a single person on freshly thawed sera that had been frozen at -20°C. RESULTS: The rate of detection of clinically significant alloantibodies (n = 154) in microtube column systems was very similar. One hundred forty-one sera (91.6%) reacted in the DiaMed-ID, 139 (90.3%) in the ReACT, 139 (90.3%) in the BioVue, and 142 (92.2%) in the Scangel. Only 117 (76.0%) of these sera reacted in the tube LISS-IAT. The detection rates for 18 antibodies of minor clinical significance (anti-M, -N, -P1, -Lea, and -Leb) varied among the test systems: DiaMed-ID, 5 (28%); ReACT, 7 (39%); BioVue, 14 (78%); Scangel, 10 (56%); and tube LISS-IAT, 6 (33%). Antibody reactivity as determined by titer and score was very similar in all microtube column systems and higher in these systems than in the tube LISS-IAT. CONCLUSION: The sensitivity of all four microtube column systems in the detection of clinically significant red cell alloantibodies was similar and was markedly superior to that of the tube LISS-IAT. An individual cost-benefit analysis should be performed in every institution to decide whether a microtube column system should be applied. If so, the antibody screen in the microtube column agglutination system should ideally be performed in advance of the crossmatch to provide time to screen for compatible units.

BACKGROUND: Megakaryocytopoiesis and platelet production are regulated by several hematopoietic growth factors. The present study focuses on the effects of automated plateletpheresis on systemic levels of different hematopoietic growth factors. STUDY DESIGN AND METHODS: Platelet count, mean platelet volume, and serum levels of thrombopoietin, erythropoietin, interleukin-1β, interleukin-6, and stem cell factor in 21 healthy donors were measured before platelet collection, after the first half of the apheresis procedure, at the end of apheresis, and on Days 1, 2, and 7 thereafter. RESULTS: Thrombopoietin levels (initial level: 49.5 ± 25.5 pg/mL) showed a significant increase between measurements taken at the end of apheresis and Day 1 (56.9 ± 26.7 pg/mL; p = 0.01). There was a highly significant decrease in stem cell factor levels during apheresis (p

BACKGROUND: Transfusions or pregnancies can cause immunization against private HLA determinants and public epitopes shared by more than one private HLA antigen. HLA antibodies are correlated with febrile transfusion reactions, lower platelet response following platelet transfusion, and an increased rate of renal transplant rejection. Until now, antibody specificities in alloantisera from platelet recipients have been poorly characterized. STUDY DESIGN AND METHODS: Consecutive serum screens from platelet recipients were analyzed for antibodies against private HLA class I antigens and public HLA epitopes using a serum analysis program based on the 2 × 2 table analysis of correlations. Serum screens of highly immunized patients and of patients with new alloimmunization events were reviewed separately. RESULTS: Of the serum screens from 566 platelet recipients, 1577 indicated alloimmunization (panel-reactive antibodies >5%). The program assigned a specificity in 1024 of these screens (64.9%) and at least once in 522 of 566 patients (92.2%). In 267 patients, antibodies detecting public epitopes in the combined A- or B-locus cross-reacting groups were found; other public markers were detected in 39 patients. Patterns of reactivity were remarkably less stable than in patient groups previously studied. In many patients, antibodies with apparent private epitope specificity preceded the identification of antibodies against a shared marker of the same cross-reactive group. However, the disappearance of antibodies (whether or not this was followed by a new antibody against a private or public marker belonging to another cross-reacting group) was also observed. CONCLUSION: The computerized analysis of microlymphocytotoxicity tests enhances the rate of antibody specification in sera from platelet recipients with lymphocytotoxic antibodies. The identified antibodies should be taken into account in the selection of platelet donors. The data confirm and extend previous observations on HLA class I antibodies and elucidate new alloimmunization events.

BACKGROUND: It is necessary to protect patients from white cell (WBC)- caused side effects of platelet transfusion by reducing the WBC contamination in single-donor platelets. STUDY DESIGN AND METHODS: A new COBE Spectra WBC (leuko)-reduction system (LRS) was compared to the COBE standard plateletpheresis (standard) procedure. Each of 62 donors underwent plateletpheresis under the two protocols (LRS and standard). The collection efficiency and WBC contamination in the components collected using the techniques were compared. Platelets were counted in a cell counter and WBCs were counted using two full grids of a Nageotte chamber. RESULTS: The preseparation and postseparation numbers for red cells, WBCs, and platelets as well as the number of collected platelets were not different in the two techniques. Collection efficiency in the LRS procedures was 96.2 +/− 13.0 percent of that in the standard procedures. Median WBC contamination in the platelet components was 10,160 per LRS procedure and 56,500 per standard procedure. The purity of the LRS components was significantly improved (p = 0.001), as seen in a comparison of the WBC numbers in components per procedure after log10 transformation (LRS: 0.096 +/− 0.195 × 10(6); standard: 0.390 +/− 1.075 × 10(6)). CONCLUSION: These data suggest that the LRS procedure produces platelet concentrates with a collection efficiency that is comparable to that obtained with the standard technique and with a residual WBC content that satisfies even the most stringent criteria for filtered platelets. As this purity can be achieved without platelet loss or alteration, conventional fiber filtration no longer seems necessary or useful in this type of single-donor platelet component.

BACKGROUND: There are no recent studies on transfusion practice and blood use with regard to diagnoses of European recipients. We conducted a survey of blood component use, including packed red cells, fresh- frozen plasma, and platelets, in an acute-care university hospital in the Greater Nurnberg area. STUDY DESIGN AND METHODS: A survey was carried out of blood component transfusion at a university hospital (Erlangen, Germany) between June 1994 and May 1996. Transfused units were listed by broad diagnostic categories formed from principal diagnoses of the recipients according to the International Classification of Diseases, Ninth Revision. RESULTS: Among 100,497 discharged patients, 6,590 patients who received transfusion (6.6%) are represented in this survey. Of 28,440 red cell units and 8,592 fresh- frozen plasma units, 72.4 percent and 66.9 percent, respectively, were used in patients with neoplastic diseases, circulatory system diseases, or disorders of the digestive system. Of 2704 platelet units, 78.1 percent were transfused to patients with neoplastic or gastrointestinal diseases or diseases of blood-forming organs. These four diagnostic categories accounted for 77.7 percent of all costs of transfusion therapy. Males received 60.1 percent of all blood components transfused, and patients less than 65 years old received 68.0 percent. CONCLUSION: This survey provides information on blood component usage in a German university hospital. It demonstrates the concentration of today's blood utilization among a few diagnostic categories. The study shows that detailed information on local blood use may be obtained quickly by using data available from transfusion services and medical record departments. This information is relevant for quality management of transfusion practice, cost analyses and for planning local and regional blood donation programs.

Our data suggest that ex vivo bone marrow purging using monoclonal antibodies (MoAbs) and sheep-antimouse immunobeads (SAM beads) prior to autologous bone marrow transplantation (ABMT) allows satisfactory tumor cell reduction without critical stem cell losses. Nevertheless, there is only a reduction but no elimination of tumor cells. The consequences will have to be clinically discussed.