The predictive value of white cell or CD34+ cell count in the peripheral blood for timing apheresis and maximizing yield

BACKGROUND: The collection of peripheral blood stem and progenitor cells (PBPCs) for transplantation can be time-consuming and expensive. Thus, the utility of counting CD34+ cells and white cells (WBCs) in the peripheral blood was evaluated as a predictor of CD34+ cell yield in the apheresis component. STUDY DESIGN AND METHODS: The WBC and CD34+ cell counts in the peripheral blood and the apheresis components from 216 collections were assessed. Sixty-three patients underwent mobilization with chemotherapy plus filgrastim, and 17 patients and 14 allogeneic PBPC donors did so with filgrastim alone. The relationship between the number of WBC and CD34+ cells in the peripheral blood and in the apheresis component was analyzed by using rank correlation and linear regression analysis. RESULTS: The correlation coefficient for CD34+ cells per liter of peripheral blood with CD34+ cell yield (x 10(6)/kg) was 0.87 (n = 216 collections). This correlation existed for many patient and collection variables. However, patients with acute myeloid leukemia had fewer CD34+ cells in the apheresis component at any level of peripheral blood CD34+ cell count. Components collected from patients with CD34+ cell counts below 10 x 10(6) per L in the peripheral blood contained a median of 0.75 x 10(6) CD34+ cells per kg. When the WBC count in the blood was below 5.0 x 10(9) per L, the median number of CD34+ cells in the peripheral blood was 5.6 x 10(6) per L (range, 1.0-15.5 x 10(6)/L). A very poor correlation was found between the WBC count in the blood and the CD34+ cell yield (p = 0.12, n = 158 collections). CONCLUSION: The number of CD34+ cells, but not WBCs, in the peripheral blood can be used as a predictor for timing of apheresis and estimating PBPC yield. This is a robust relationship not affected by a variety of patient and collection factors except the diagnosis of acute myeloid leukemia. Patients who undergo mobilization with chemotherapy and filgrastim also should undergo monitoring of peripheral blood CD34+ cell counts, beginning when the WBC count in the blood exceeds 1.0 to 5.0 x 10(9) per L.     

Allogeneic blood progenitor cell collection in normal donors after mobilization with filgrastim: the M.D. Anderson Cancer Center experience

BACKGROUND: Information on the safety and efficacy of allogeneic peripheral blood progenitor cell (PBPC) collection in filgrastim-mobilized normal donors is still limited. STUDY DESIGN AND METHODS: The PBPC donor database from a 42-month period (12/94-5/98) was reviewed for apheresis and clinical data related to PBPC donation. Normal PBPC donors received filgrastim (6 microg/kg subcutaneously every 12 hours) for 3 to 4 days and subsequently underwent daily leukapheresis. The target collection was > or =4 x 10(6)CD34+ cells per kg of recipient's body weight. RESULTS: A total of 350 donors were found to be evaluable. Their median age was 41 years (range, 4-79). Their median preapheresis white cell count was 42.8 x 10(9) per L (range, 18.3-91.6). Of these donors, 17 (5%) had inadequate peripheral venous access. Leukapheresis could not be completed because of apheresis-related adverse events in 2 donors (0.5%). Of the 324 donors evaluable for apheresis yield data, 221 (68%) reached the collection target with one leukapheresis. The median CD34+ cell dose collected (first leukapheresis) was 462 x 10(6) (range, 29-1463).The main adverse events related to filgrastim administration in donors evaluable for toxicity (n = 341) were bone pain (84%), headache (54%), fatigue (31%), and nausea (13%). These events were rated as moderate to severe (grade 2-3) by 171 (50%) of the donors. In 2 donors (0.5%), they prompted the discontinuation of filgrastim administration. CONCLUSION: PBPC apheresis for allogeneic transplantation is safe and well tolerated. It allows the collection of an "acceptable" PBPC dose in most normal donors with one leukapheresis, with minimal need for invasive procedures.     

Donor age-related differences in PBPC mobilization with rHuG-CSF

BACKGROUND: Data on the administration of rHuG-CSF to normal donors <18 years old are very limited. STUDY DESIGN AND METHODS: The results of rHuG-CSF administration to 61 donors <18 years old (Group A) were retrospectively evaluated and compared with results from 353 donors > or = 18 years old (Group B) who are included in the Spanish National Donor Registry. The mean age (range) in Group A and B was 14 (1-17) and 38 (18-71) years, respectively (p<0.001). The mean dose of rHuG-CSF was 10 microg per kg per day (range, 9-16) during a mean of 5 days (range, 4-6). Central venous access was placed more frequently in younger donors (25% vs. 6%; p<0.001). RESULTS: The mean number of CD34+ cells collected was 7.6 and 6.9 x 10(6) per kg of donor's body weight in Group A and B, respectively. Fifty-six percent of Group A donors needed only one apheresis to achieve > or = 4 x 10(6) CD34+ cells per kg versus 39 percent of Group B donors (p = 0.01). Side effects were more common in Group B (71% vs. 41%; p<0.001). CONCLUSION: The administration of rHuG-CSF to donors <18 years old leads to CD34+ cell mobilization in a pattern similar to that observed in adults. Greater age was associated with a more frequent requirement for more than one apheresis to achieve a similar number of CD34+ cells.     

Safety of growth factor administration for leukapheresis in those with WBC counts greater than 60,000/µl

Purpose: Peripheral blood stem cell mobilization using growth factors is a common method of stem cell collection for transplantation, however, little is reported concerning safety of continued growth factor delivery in exceptional responders with very high white blood cell (WBC) counts in preparation for pheresis. We performed a retrospective study of the safety of growth factor delivery for leukapheresis in those with WBC counts greater than 60,000/µl. Methods: Allogeneic donors received 5 days of granulocyte colony‐stimulating factor (G‐CSF) at a daily dose of 10 or 16 µg/kg. Autologous donors received G‐CSF 10 µg/kg/day +/? chemotherapy until peripheral blood CD34⁺count reached 10/µl. Granulocyte donors received 300 µg dose of G‐CSF the day prior to donation. Results: Out of 3,037 leukapheresis collections from 1998 to 2005, we identified 303 collections from 204 donors or patients who had a WBC > 60,000/µl. WBC counts were ≥100,000/µl in seven of these subjects. If inadequate stem cell dose was obtained with pheresis with WBC counts this high, patients had growth factor dosing decreased 50% but still received a dose till stem cell collection was completed. Of the 204 subjects, 122 were patients and 82 were donors. These 204 donors/patients had no serious adverse events reported other than the common reports of myalgia, bone pain, and headache associated with administration of growth factors. Pain levels ranged from mild to severe and usually were managed by over the counter analgesics. Conclusions: Continuing ½ the dose of neupogen to complete the pheresis process appears safe in subjects with very high white blood counts. J. Clin. Apheresis 30:28–31, 2015. © 2014 Wiley Periodicals, Inc.     

Administration of G--CSF plus dexamethasone produces greater granulocyte concentrate yields while causing no more donor toxicity than G--CSF alone

BACKGROUND: G-CSF with or without dexamethasone is becoming the standard agent for mobilizing granulocytes for transfusion. The purpose of this study was to determine if the toxicities of G--CSF with or without dexamethasone are offset by greater collection yields and to define the minimum interval that should separate sequential collections. STUDY DESIGN AND METHODS: Twenty donors were studied on three occasions. They were given either dexamethasone (8 mg, by mouth) plus a placebo injection, G--CSF (5 microg/kg, given subcutaneously) plus placebo capsules, or G--CSF plus dexamethasone. Granulocytes were collected by apheresis. A donor symptom survey was administered, and cell counts and blood chemistries were assessed before collection and 1, 2, 7, 14, 21, 28, and 35 days after collection. RESULTS: More granulocytes were collected when G--CSF was given than when dexamethasone was given (41.1 +/- 20.4 x 10(9) vs. 21.0 +/- 10.0 x 10(9); p<0.001), but the use of G--CSF plus dexamethasone produced the greatest yields (67.1 +/- 22.0 x 10(9); p<0.002). When the donors were given dexamethasone alone, 58 percent experienced at least one symptom, compared to 85 percent of those given G--CSF and 75 percent of those given G--CSF plus dexamethasone. In all three regimens, platelet counts fell 19 percent to 24 percent after collection and remained below baseline for 7 to 14 days. Granulocyte counts returned to baseline within 3 to 7 days, but, in all three regimens, a mild granulocytopenia occurred 21 days after collection. With each of the regimens, blood chemistries changed, but the changes were mild and most returned to baseline within 7 days; however, changes in albumin, bilirubin, and AST persisted until 28 days after collection. CONCLUSION: These results support the use of G--CSF plus dexamethasone in granulocyte donors. G--CSF plus dexamethasone resulted in greater granulocyte yields than either agent alone and was associated with donor symptoms and changes in blood cell counts and chemistries similar to those seen with G--CSF alone or dexamethasone alone. Granulocytes can be safely collected a second time after a 7-day interval; however, for regular donors, it may be best to separate collections by 4 weeks.     

Effects of different concentrations of anticoagulant on the in vitro characteristics of autologous whole blood

BACKGROUND: Routinely, 450 mL of blood is collected into 63 mL of CPDA-1, for a final anticoagulant:blood ratio of approximately 1:7 in a whole-blood autologous unit. If less than 300 mL of blood is to be collected, the AABB standards suggest that there should be a proportionate decrease in anticoagulant. Data from an autologous blood program showed a range in volume from 92 mL to 667 mL per bag, which reflects an anticoagulant:blood ratio of 2:1 to 1:10. STUDY DESIGN AND METHODS: To determine the effects of these ratios on the in vitro function of RBCs at various anticoagulant ratios, blood was collected into different amounts of anticoagulant, and various measurements were made during storage. RESULTS: The number of RBCs and the MCV remained constant over time, regardless of the anticoagulant dilution used. Plasma free Hb increased with time with all dilutions. At a 1:2 ratio, it rose from 734 mg per L on Day 1 to 1805 mg per L on Day 35, and at 1:8, it was 355 mg per L for Day 1 and 854 mg per L on Day 35. Plasma sodium decreased and the potassium increased over time with all dilutions. From Day 1 to Day 35, there was a nine-fold increase in potassium at both the 1:2 and 1:8 dilutions (2.4 to 22.9 mmol/L, 3.2 to 29.6 mmol/L, respectively). The LDH increased over time and the pH decreased in all of the dilutions. Osmotic fragility remained constant at the 1:8 dilution but decreased at all of the other dilutions with storage, with 44-percent fragility on Day 35 at the 1:2 ratio. The WBC and platelet counts decreased consistently over time. Overall, 1 percent of the autologous units were below the cutoff volume of 300 mL at which an adjustment of the anticoagulant volume is required. CONCLUSION: Plasma Hb and plasma potassium concentrations are considerably higher in low-volume units, which indicates that deviation from standard collection procedures is deleterious to RBCs.     

The acquisition of granylocytes by leukapheresis: a comparison of continuous flow centrifugation and filtration leukapheresis in normal and corticosteroid-stimulated donors

Leukapheresis by continuous flow centrifugation (CFC) or filtration (FL) were compared in untreated and corticosteroid-treated donors. The administration of prednisone 60 mg orally 10 to 12 hours before leukapheresis increased significantly both the donor's prepheresis WBC counts and the total granulocyte yields by CFC or FL. Dexamethasone 6 mg iv at the start of FL did not increase granulocyte yields significantly. In untreated donors FL yielded 0.25 × 10(10) granulocytes per liter donor blood processed as compared with 0.05 × 10(10) per liter in CFC donors. Corticosteroid premedication has a greater relative effect in increasing yields by CFC than by FL. Donor reaction rates were between 4 and 6 per cent for both procedures.     

Analysis of factors associated with low peripheral blood progenitor cell collection in normal donors

BACKGROUND: Predictive factors of the response to rHuG-CSF in normal donors have not been extensively studied. STUDY DESIGN AND METHODS: We analyzed factors influencing CD34+ cell yield in the 1st day of collection in 261 healthy donors from the Spanish National Donor Registry. The median age was 38 years (range, 2-72). The median dose of rHuG-CSF was 10 microg per kg per day (range, 5-20) over 4 days. In 103 donors (40%), <4 x 10(6) per kg CD34+ cells were collected. The variables that were analyzed included age, sex, weight, basal complete blood cell count, dose, type of rHuGCSF and schedule of administration, and maximum WBC count before apheresis. RESULTS: By univariate analysis, the maximum WBC count (<50 vs. >or=50 x 10(9)/L, p = 0.004), advanced age (p = 0.008), and number of daily rHuG-CSF doses (one vs. two; p = 0.01) correlated with the number of CD34+ cells collected. By multivariate analysis, donors age (<38 vs. >or=38 years; p = 0.014) and a single daily dose of rHuG-CSF (p = 0.005) were the two variables that significantly predicted a low CD34+ cell yield. CONCLUSION: Donors' age, with a threshold of 38 years or more, and the rHuG-CSF schedule are the factors that significantly affected CD34+ cell mobilization and collection in healthy donors.     

Combined administration of G-CSF and dexamethasone for the mobilization of granulocytes in normal donors: optimization of dosing

BACKGROUND: The clinical utility of neutrophil (polymorphonuclear leukocyte, PMN) transfusion therapy has been compromised, in part, by the inability to obtain sufficient quantities of functional neutrophils from donors. Mobilization of PMNs in the peripheral blood of normal volunteers has been shown to be superior when G-CSF is administered in conjunction with dexamethasone to that when either agent is administered alone. The current study was conducted to determine the optimal dosages of G-CSF and dexamethasone to be administered to donors in a granulocyte transfusion program. STUDY DESIGN AND METHODS: Five normal subjects were randomly assigned to each of the following single-dose regimens over five consecutive weeks: 1) subcutaneous (SC) G-CSF at 600 microg and oral (PO) dexamethasone at 8 mg; 2) SC G-CSF at 450 microg and PO dexamethasone at 8 mg; 3) SC G-CSF at 450 microg and PO dexamethasone at 12 mg; 4) SC G-CSF at 450 microg; and 5) PO dexamethasone at 12 mg. Venous blood was collected at 0, 6, 12, and 24 hours after drug administration for determination of absolute neutrophil count (ANC). Side effects of drug administration were recorded by using a standardized symptom questionnaire. RESULTS: Maximal ANC was achieved at 12 hours after administration of drugs under each regimen. All four regimens containing G-CSF caused greater than 10-fold increases in the ANC. When administered in conjunction with dexamethasone, G-CSF resulted in statistically similar PMN mobilization at dosages of 450 microg and 600 microg. The combined single-dose regimen of SC G-CSF at 450 microg and PO dexamethasone at 8 mg increased the mean ANC from a baseline value of 2800 per microL to 37,900 per microL at 12 hours after administration. This regimen was well tolerated by the normal volunteers. CONCLUSION: In a single-dose format designed for clinical granulocyte transfusion programs, optimal PMN mobilization can be achieved in normal donors with a combined regimen of SC G-CSF at 450 microg, and PO dexamethasone at 8 microg.     

Effects of granulocyte–colony-stimulating factor on potential normal granulocyte donors

BACKGROUND: The use of granulocyte-colony-stimulating factor (G-CSF) to increase the granulocyte count and the yield from leukapheresis in normal donors is leading to renewed interest in granulocyte transfusion. Therefore, it is important to understand the side effects of G-CSF. STUDY DESIGN AND METHODS: We studied the effect of G-CSF on peripheral blood counts and recorded the side effects experienced 24 hours after an injection of G-CSF in normal subjects donating peripheral blood progenitor cells for research. RESULTS: Following administration of G-CSF to 261 donors, the neutrophil count increased to 20.6 to 24.5 x 10(9) per microL depending on the dose of G-CSF. This represented a 6.2 to 7.4-fold increase over the neutrophil count before G-CSF administration. Of all donors, 69 percent experienced one or more side effects. The most common effects were: muscle and bone pain, headache, fatigue, and nausea. There was a relationship between the dose of G-CSF and the likelihood of experiencing a side effect. Most side effects were mild, but about 75 percent of donors took analgesics because of them. CONCLUSIONS: In a granulocyte donation program involving G-CSF stimulation, about two-thirds of donors would experience one or more side effects, but these would usually be mild and well tolerated.     

WBC content of platelet concentrates prepared by the buffy coat method using different processing procedures and storage solutions

BACKGROUND: The number of WBCs in platelet concentrates (PCs) prepared by the buffy coat (BC) method with different storage solutions can result in low (5 x 10(6)/unit) WBC levels by the use of careful centrifugation techniques without filtration. At present, most blood banks use filtration steps to meet these requirements. The difference in processing methods and suspension solutions prompted the investigation of the influence of the various procedures on the WBC and platelet content of PCs. STUDY DESIGN AND METHODS: PCs from 5 BCs were harvested without or with inline filtration (AutoStop BC, Pall Corp.) in either plasma (PCs-plasma) or platelet additive solution (PCs-PAS-2). After preparation, samples were taken for counting WBCs and platelets and for analyzing WBC subsets by flow cytometry using specific MoAbs. The WBCs were concentrated before analysis of the WBC subsets. Results less than 2.5 cells per microL were considered below the limit of accuracy of the subset analysis. RESULTS: All filtered PCs met the AABB standard of 5 x 10(6) per unit and the European guidelines of 1 x 10(6) per unit. None of the nonfiltered PCs met the European guidelines, but all met the AABB guidelines. All filtered units gave residual WBC counts below the detection limit for subset analysis. Filtered PCs-plasma gave significantly higher platelet counts than filtered PCs-PAS-2 or nonfiltered PCs (p<0.01, ANOVA). CONCLUSION: Careful centrifugation of pooled BCs, with plasma or PAS-2, can result in PCs with low WBC contamination levels. However, filtered PCs are superior, because of better WBC removal and higher platelet counts.     

The determinants of granulocyte yield in 1198 granulocyte concentrates collected from unrelated volunteer donors mobilized with dexamethasone and granulocyte–colony-stimulating factor: a 13-year experience

BACKGROUND: The combination of granulocyte–colony-stimulating factor (G-CSF [filgrastim]) and dexamethasone (G-CSF/dex) is an effective granulocyte mobilization regimen, but the variables that affect donor neutrophil response and granulocyte collection yield are not well characterized.
STUDY DESIGN AND METHODS: A computerized database containing records of 1198 granulocyte collections from 137 unrelated volunteer apheresis donors during a 13-year period was retrospectively analyzed. Donors were categorized by age, sex, and cumulative number of granulocyte donations. Complete blood counts at baseline and after G-CSF/dex stimulation were recorded. The outcome variables include the preprocedure absolute neutrophil count (preANC), which reflects G-CSF/dex stimulation, and the granulocyte product yield per liter processed (BagGranYield/L).
RESULTS: Higher baseline ANC and platelet (PLT) counts were significantly associated with higher preANC while a larger number of prior granulocytapheresis procedures was associated with lower preANC. Total filgrastim dose (used in weight-based dosing) did not significantly impact preANC or the granulocyte yield; weight-based dosing at 5 µg per kg and a uniform 480-µg dose produced equivalent preANC. PreANC and weight were the key determinants of granulocyte yield (BagGranYield/L).
CONCLUSION: Apheresis donors with higher baseline PLT counts and ANCs have higher ANCs after G-CSF/dex stimulation; donor age, weight, and sex do not have a significant impact. A uniform G-CSF dose of 480 µg is as effective as weight-based dosing at 5 µg per kg. Donor ANC monitoring should be considered after serial granulocytapheresis procedures.     

White cell reduction during plateletpheresis: a comparison of three blood cell separators

BACKGROUND: White cell (WBC)-reduced single-donor platelet concentrates (SDPs) can be collected by the newest generation of blood cell separators. Three WBC-reduction techniques during plateletpheresis were investigated in the present study with respect to WBC content and platelet yield. STUDY DESIGN AND METHODS: The Amicus device used the elutriation principle for WBC reduction, and separations with periodically alternating interface position (PAIP) were employed in the AS.TEC 204. WBC reduction by in-line filtration was performed in the MCS+. Platelets were measured electronically and WBCs were determined manually (Nageotte chamber). RESULTS: In-line filtered SDPs showed significantly lower WBC content (0.088+/-0.178 x 10(6)) than SDPs that were WBC reduced by elutriation (0.31+/-0.48 x 106) or PAIP technique (0.89+/-1.57 x 10(6), p = 0.0001). Platelet yield (5.0+/-0.46 x 10(11)) was significantly higher in components obtained with the Amicus device (p = 0.0001). The AS.TEC 204 and MCS+ gave similar results for platelet yields: 3.15+/-0.63 and 3.28+/-0.71 x 10(11), respectively. CONCLUSIONS: The plateletpheresis systems studied allow the collection of WBC-reduced SDPs. In-line filtration resulted in the best WBC reduction. Some SDPs collected with the devices studied had a WBC content >1 x 10(6) per unit. Platelet yield was significantly higher in SDPs from the Amicus device.     

Granulocyte harvest for transfusion: donor response to repeated leukapheresis

Leukapheresis of normal donors with the NCI-IBM Continuous Flow Blood Cell Separator was compared with the method of filtration leukapheresis. An average of 5.7 times 10-9 (range 1.4 to 10.9 times 10- 9) granulocytes were collected on 25 occasions with the blood cell separator compared with an average of 36.6 times 10-9 (range 20.1 to 61.3 times 10-9) obtained by filtration leukapheresis on 85 occasions. Donor platelet counts decreased following donation by an average of 29,000/mul (14%) with continuous flow centrifugation (CFC) and an average of 40,000/mul (17%) with filtration leukapheresis (FL). Estimated donor packed red blood cell loss, including blood obtained for investigative purposes, was 80 ml per donation with CFC and 67 ml per donation when FL was used. FL resulted in at least a 25 per cent increase in the granulocyte count of donors by donation completion in 61 per cent of donors, although postdonation granulocytosis did not correlate with increased cell harvest. CFC donors generally displayed a postdonation decrease in granulocyte count. Normal numbers of granulocytes were maintained in donors undergoing frequent leukapheresis and no donor's health was compromised by the removal of large numbers of granulocytes. Repeated donations were possible with both systems. Although platelet counts dropped more with FL, the procedure was performed as frequently as eight times in a ten-day period without development of severe donor thrombocytopenia. Donor red blood cell loss appears to be the limiting factor in the repeated use of either system. Blood loss could be significantly decreased under routine conditions if fewer samples were obtained for investigational purposes. Filtration leukapheresis offers the advantage of significantly greater granulocyte yields than is possible with CFC unless modifications such as arterio-venous shunts or the use of steroids and/or rouleaux-inducing agents are employed.     

Evaluation of a concurrent multicomponent collection system for the collection and storage of WBC-reduced RBC apheresis concentrates

BACKGROUND: Multicomponent apheresis procedures offer the possibility of collecting blood components that are standardized, as compared to those available with whole-blood donations. A new separator program for the concurrent collection of RBCs, platelets, and plasma (Amicus, Baxter Healthcare) was evaluated. STUDY DESIGN AND METHODS: Apheresis donors (n = 47) underwent concurrent collection of RBCs, platelets, and plasma by use of the single-needle procedure of the Amicus blood cell separator. A standardized RBC volume (100% Hct) of 200 mL was targeted with either 1 or 2 platelet concentrate units, depending on the donor's predonation characteristics. After collection, the RBC component was sterilely connected to an RBC collection set (Amicus) to allow for the addition of 100 mL of saline-adenine-glucose-mannitol preservative solution and WBC reduction at either ambient temperature or 4 degrees C. The RBC units were subsequently stored at 2 to 6 degrees C for 42 days, and the following in vitro measures were evaluated over the storage period: blood cell counts including Hct and total Hb, plasma Hb, potassium, pH, ATP, and 2,3 DPG. RESULTS: Procedure time averaged 74 +/- 9 minutes, and no adverse events were reported. The absolute RBC volume collected averaged 198 +/- 11 mL with an average Hct value of 83 +/- 2 percent. After filtration, the Hb content averaged 58.2 +/- 2.4 g per unit and residual WBCs averaged 0.038 +/- 0.015 x 10(6) per unit. Day 42 results showed that all units had on average more than 70-percent ATP maintenance, and all of the units had less than 0.8 percent he-molysis. All units had pH values higher than 6.5 on Day 42. CONCLUSION: The concurrent multicomponent collection system (Amicus) can reliably collect a standardized RBC unit of good quality. In vitro testing of the RBCs collected and stored for 42 days met the Council of Europe criteria for transfusion.     

'In vivo' time course of plasma myeloperoxidase levels after granulocyte colony-stimulating factor-induced stem cell mobilization

Administration of granulocyte colony-stimulating factor (G-CSF) is widely used for harvesting an adequate number of CD34+ stem cells by leukapheresis in normal donors. G-CSF is the most established agent for the mobilization of stem cells in current clinical practice, because it has been proven to be superior to any other agent tested to date in terms of not only mobilization capacity, but also of tolerance. However, although regulatory and accrediting agencies have provided guidelines to protect donors, the short- and long-term side effects of G-CSF need to be further studied. In this study, we evaluated the time course of plasma myeloperoxidase (MPO) levels measured in a group of donors given recombinant human G-CSF (rHuG-CSF) at different intervals: (i) before starting rHuG-CSF administration, (ii) on day 5 of rHuG-CSF administration, (iii) on the same day soon after the end of the first leukapheresis procedure and (iv) 1 week after rHuG-CSF withdrawal. Plasma MPO levels significantly increased in the donors after 5 days of rHuG-CSF treatment, returning to the baseline values within 7 days following rHuG-CSF withdrawal. These findings may contribute to a better understanding of G-CSF safety profile in stem cell donors.     

Comparison of intermittent- and continuous-flow cell separators for the collection of autologous peripheral blood progenitor cells in patients with hematologic malignancies

BACKGROUND: The transplantation of autologous peripheral blood progenitor cells (PBPCs) after high-dose chemotherapy is a valuable therapy for patients with hematologic and solid malignancies. Several methods are used for harvesting PBPCs. The efficiency of intermittent- and continuous-flow blood cell separators in collecting progenitor cells from the blood of patients undergoing myeloablative treatment for cancer was compared. STUDY DESIGN AND METHODS: PBPC components (n = 133) were obtained from 72 patients by leukapheresis with continuous-flow machines (Spectra, COBE; CS 3000 Plus, Baxter) and with an intermittent-flow machine (MCS 3P, Haemonetics). The data were analyzed retrospectively. Blood samples obtained from the patients before leukapheresis and samples of the leukapheresis components themselves were analyzed for their content of RBCs, WBCs, platelets, and CD34+ cells. RESULTS: The Spectra processed more than twice the blood volume in the shortest time (15 L in 178 min), whereas the Baxter CS 3000 Plus (10 L in 185 min) and the MCS 3P (4.8 L in 239 min) processed significantly smaller volumes in a longer time. The mean ACD consumption was 403 mL with the MCS 3P, 900 mL with the CS 3000 Plus, and 1000 mL with the Spectra. The product volumes were 50 mL (CS 3000 Plus), 69 mL (MCS 3P), and 166 mL (Spectra). In all groups, differences in the preapheresis hemograms were not significant, but the Spectra group had fewer CD34+ cells than the other groups. Despite this, the differences in the number of CD34+ cells in the leukapheresis components of all groups were without statistical significance. In the Spectra group, the collection of MNCs of 104 percent and CD34+ cells of 154 percent was significantly more efficient than that in the MCS 3P group (42.2% and 56%, respectively) or the CS 3000 Plus group (50.8% and 47.15%) as related to the patients' blood volume. CONCLUSION: PBPC collection can be performed successfully with continuous-flow and intermittent-flow blood cell separators. The Spectra had the best recovery of CD34+ cells within the shortest time. Leukapheresis with the MCS 3P is indicated if only a single venous access is available.     

Evaluation of a whole-blood WBC-reduction filter that saves platelets: in vitro studies

BACKGROUND: In this study, a new WBC-reduction in-line filter that removes WBCs but not platelets was evaluated. Three WBC-reduced blood components were prepared: RBCs, plasma, and platelet concentrates (PCs). STUDY DESIGN AND METHODS: Whole-blood components (n = 30) were filtered within 2 to 4 hours after collection and then were centrifuged and separated into RBCs, plasma, and WBC-reduced buffy coat. Saline-adenine-glucose-mannitol solution was added to the RBCS: The WBC-reduced buffy coats were stored overnight; on the following day, PCs were prepared from pooled WBC-reduced buffy coats and stored in a medium composed of approximately 35 percent CPD plasma and 65 percent platelet additive solution (T-Sol, Baxter). The WBC-reduction capacity of the filter, the recovery of cells after filtration, and the in vitro storage of RBCs (n = 10) and platelets (n = 6) were evaluated. RESULTS: Mean and maximum WBC counts after filtration were 0.08 x 10(6) and 0.3 x 10(6), respectively, per filtered whole-blood unit. Recovery of RBCs (mean values) after filtration was 90 percent in whole-blood components and 73 percent in RBCS: Recovery of platelets (mean values) was 81 percent after filtration and 66 percent in PCS: The in vitro storage study of RBCs showed results comparable with previously published data, except for a lower degree of hemolysis. In the in vitro platelet storage study, results were compared with those of standard preparations. In all essentials, similar results were found. CONCLUSION: The results of the present study suggest that effective WBC reduction meets current standards and satisfactory recovery after filtration. The storage characteristics for RBCs and PCs are similar to those of standard preparations. Use of a whole-blood in-line filter to save platelets is a new option for whole-blood processing, which may simplify WBC reduction and blood component preparation, as well as reduce costs in the future.     

Residual subset population analysis in WBC-reduced blood components using real-time PCR quantitation of specific mRNA

BACKGROUND: Implementation of WBC reduction of the blood supply increases the importance of measurement of residual WBC subtypes responsible for immunologic and infectious complications of transfusion. STUDY DESIGN AND METHODS: Real-time RT-PCR assays were developed to detect mRNA encoding lineage-specific WBC markers. Primers and fluorescent probes were designed for CD45 (pan-WBC), CD3 (T-lymphocyte), CD19 (B-lymphocyte), CD14 (monocyte), and CD66 (granulocyte), and the specificity was assessed by comparison with flow cytometric analysis of enriched cell populations. WBC subsets were examined in WBC-reduced whole blood prepared with filters (WBF2, Pall; and RZ2000, Baxter) and in platelet concentrates prepared with other filters (Autostop, Pall; and PLX-5, Baxter) and apheresis (COBE Spectra LRS, Gambro). RESULTS: All real-time RT-PCR assays were linear over >5 log concentration range, allowing pre-WBC-reduction and post-WBC-reduction comparisons. Sensitivity limits ranged from 10 cells per mL (CD45) to 200 cells per mL (CD19). Assay specificity was confirmed by the close correlation of real-time RT-PCR and immunophenotyping results by flow cytometry. For all subsets, >3.8 log and >3.1 log reduction was obtained during WBC reduction of whole blood and platelets, respectively. CONCLUSION: Real-time RT-PCR assays are suitable for analysis of subset removal during WBC reduction. There was no significant difference between the two whole-blood filters or between platelet filtration and apheresis in the removal of any WBC subset.     

The dose of granulocyte–colony-stimulating factor after chemopriming treatment does not influence apheresis yield of progenitor cells: a retrospective study of 91 cases

BACKGROUND: The optimal dose of post-chemotherapy granulocyte–colony-stimulating factor (G–CSF) administration before peripheral blood progenitor cell (PBPC) collection has not been determined as yet, although 5 μg per kg per day has been recommended as the standard dose. This study retrospectively analyzed the effect of G–CSF dose on peripheral blood CD34+ cell collection from 91 patients with hematologic malignancies.
STUDY DESIGN AND METHODS: Various doses of G–CSF were administered after several chemotherapeutic PBPC mobilization regimens. According to the dose of G–CSF administered, patients were assigned to two groups. Group 1 included 46 patients who received a low dose of G–CSF (median, 3.6 [range, 2.8-4.6] μg/kg/day). Group 2 included 45 patients who received a standard G–CSF dose of 6.0 (5.5-8.1) μg per kg per day. Patients in the two groups were matched for age, diagnosis, previous therapy, and chemotherapeutic PBPC mobilization regimens.
RESULTS: No difference was observed in the median number of CD34+ cells harvested from each group. The number of leukapheresis procedures necessary to obtain a minimum of 3 × 10⁶ CD34+ cells per kg was the same in both groups, and the percentage of patients who failed to achieve adequate PBPC collections was similar in the two groups.
CONCLUSION: The administration of low-dose G–CSF after chemotherapy appears equivalent to administration of the standard dose in achieving satisfactory PBPC collection. This approach could allow significant savings in medical cost. A randomized and prospective study is necessary, however, to assess the validity of these conclusions.