Bone Marrow Transplantation (2000) 25, 371–376  2000 Macmillan Publishers Ltd All rights reserved 0268–3369/00 $15.00 www.nature.com/bmt Lenograstim-mobilized peripheral blood progenitor cells in volunteer donors: an open label randomized split dose escalating study

N Basara, B Schmetzer, IW Blau, M Bischoff, S Gu¨nzelmann, D Kirsten and AA Fauser

Clinic of Bone Marrow Transplantation and Haematology/Oncology and European Institute for Research and Development of Transplant Strategies, Idar-Oberstein, Germany

Summary: 1000 PBSCT were reported to the European Group for Blood and Marrow Transplantation (EBMT) indicating a Mobilization of peripheral blood cell progenitor cells dramatic increase in the number of donors undergoing stem was investigated in 36 healthy sibling donors using three cell mobilization.6 Collection of PBSC with a limited num- different split doses of glycosylated rhG-CSF ber of leukaphereses requires mobilization with haematopo- (lenograstim). The donors were randomized into three ietic growth factors. It has been shown that short-term groups: group 1 was given lenograstim at 8, group 2 administration of rhG-CSF does allow the harvest of a large at 11 and group 3 at 15 ␮g/kg/day in two split doses, number of PBSC from healthy volunteer donors.7,8 How- subcutaneously for 4 and 5 days, respectively. Leukaph- ever, the optimal dosage for CD34ϩ cell mobilization is not eresis was performed on day 4 or 5 depending on the yet defined.9 Based on earlier reported data9–11 the most WBC and CD34؉ cell count. We were able to demon- favourable time for stem cell collection might be day 4 to strate that there was a significant correlation between 5 after cytokine treatment. Continuation of rhG-CSF circulating CD34؉ cells on the day of harvest and administration beyond a 5-day course led to a rapid decline CD34؉ cells in the apheresis products in all three in the mobilization efficiency of CD34ϩ cells.11 It has been groups. The number of CD34؉ cells pre-apheresis was demonstrated that, at least for rhG-CSF at doses between inversely correlated with age in group 1 and group 2. 5 and 10 ␮g/kg/day, a dose-response relationship does exist -However, in group 3, the number of CD34؉ cells pre- with regard to the yield of CD34ϩ cells.12 Although rhG apheresis did not correlate with age. There was also a CSF doses up to 24 ␮g/kg/day have been reported,13 experi- difference between the number of progenitor cells mobi- ence with such high amounts remains limited. The purpose lized in the three dose groups regarding the time of har- of this dose escalating study was to determine whether vest. Apheresis was performed in groups 1 and 2 on day escalating split doses of lenograstim ranging from 8 to 15 5 of mobilization in order to obtain a sufficient number ␮g/kg do result in an increase in the number of CD34ϩ of stem cells for allogeneic transplantation. In contrast, cells harvested and whether higher doses of G-CSF might with the split dose of 15 ␮g/kg/day, harvest could be allow collection earlier. routinely performed on day 4 of stimulation. We con- clude that lenograstim given twice a day at doses of 8, ؉ 11 and 15 ␮g/kg/day provided different CD34 cell Donors and methods yields in normal donors, in particular, with regard to ؉ the time of harvest. The number of CD34 cells pre- Donors apheresis was not correlated with age in the group of donors mobilized with a split dose of 15 ␮g/kg/day, indi- Thirty-six healthy donors underwent stem cell mobilization cating that this dosage might also be suitable for older between January 1998 and February 1999 at the Clinic of donors. Bone Marrow Transplantation (2000) 25, 371– Bone Marrow Transplantation and Haematology. All 376. donors were related to the recipients, 34 were siblings and Keywords: lenograstim; mobilization; allogeneic stem one was a daughter, who donated at two different times. cell transplant; volunteer donors Evaluation included full medical history and physical examination. Investigations comprised complete blood counts, renal and liver profiles, ECG and chest X-ray. Donors were randomized to receive a dose of 8 ␮g/kg, 11 Allogeneic peripheral blood stem cell transplantation ␮g/kg or 15 ␮g/kg/day. Lenograstim (Granocyte, Rhoˆne- (PBSCT) from sibling donors is increasingly used as an Poulenc Rorer, Frankfurt, Germany) was administered sub- alternative source of haematopoietic stem cells rather than cutaneously daily in two split doses for 5 days or for 4 days, 1–5 bone marrow-derived stem cells. In 1996, more than respectively. With regard to donor characteristics there was no statistically significant difference between the three groups of donors concerning age, weight and sex distri- Correspondence: Dr N Basara, Clinic of Bone Marrow Transplantation and Haematology/Oncology, Dr. Ottmar-Kohler Str. 2, 55743 Idar- bution (Table 1). In addition, no difference was found in Oberstein, Germany baseline WBC count, harvested blood volume and the dur- Received 13 May 1999; accepted 25 October 1999 ation of apheresis. The protocol used in this study was Dose of lenograstim for mobilization of PBSC N Basara et al 372 Table 1 Donor characteristics

Group 1 Group 2 Group 3

Dose (␮g/kg/day) 8.09 ± 0.18 11.02 ± 0.22 15.0 ± 0.16 No. of donors 10 14 12 Age (range) 34 (17–44) 39 (25–69) 47 (17–62) Weight 83 (48–105) 73 (56–100) 76 (53–100) Sex Female/Male 7/3 8/6 5/7 1. PBSCC on day 5 5 4 WBC (109/l) Baseline 4.8 ± 1.4 6.1 ± 1.8 5.7 ± 1.5 Pre-apheresis 46.7 ± 3.53 48.8 ± 3.63 47.0 ± 3.76 CD34+ (%) cells in PB Pre-apheresis 0.24 ± 0.02 0.20 ± 0.03 0.15 ± 0.02 CD34+ (106/l) cells in PB Pre-apheresis 109 ± 11.12 95.7 ± 12.8 70.4 ± 15.9

Age is expressed as median (range). Values are expressed as mean ± s.e.m.

approved by the IRB of the Clinic of BMT and Measurement of CD34ϩ cells Haematology/Oncology. The procedure, as well as the risks and benefits were explained in detail to the donors. All PBSC products had a 1 ml sample removed and diluted 1:10 in autologous plasma. Routine analysis of these samples included a WBC, total nucleated cell count, pro- Safety genitor cell determination and CD34 antigen analysis. The There was a total of 65 adverse events. Bone pain was the peripheral blood samples and leukapheresis products were most frequently reported symptom (42 events). In addition, analysed by double-colour flow cytometry as previously described.14 Briefly, 1 ϫ 106 cells were stained simul- mild headache was reported in 23 donors. None of the − donors had any serious adverse events. One donor experi- taneously with the phycoerythrin (PE)-conjugated CD34 enced a transient WHO grade I increase in S-ALAT on day (HPCA-2)-antibody (Becton Dickinson, San Jose, CA, 5 of mobilization. Transient WHO grade I increase in LDH USA) and anti-CD45 (Becton Dickinson). Using the gating and AP was found in 29 donors. strategies according to the ISHAGE guidelines enumeration of the CD34ϩ cells was performed on an EPICS XL (Coulter). Subsets of CD34ϩ cells were determined by the Harvesting of PBSC expression of CD33, CD38 or HLA-DR antigens. The per- centage of CD34ϩ cells was converted to absolute numbers Following twice daily lenograstim administration, apheresis by multiplication with the leukocyte count in the sample was scheduled and performed at day 4 or day 5 (Table 1) obtained from the haematocytometer. Results were finally using a continuous flow centrifuge (Fenwal CS-3000 Plus; presented as the number of circulating CD34ϩ cells per ␮l Baxter, Unterschleißheim, Germany) for 3 to 4 h sessions peripheral blood or as the number of CD34ϩ cells per leu- ϫ processing 3 blood volume. The median processed blood kapheresis product. T and NK cells were enumerated using volume was 15 l (range: 8–16 l). The study was designed the CD3-FITC, CD16,56-PE (Immunotech, Marseille, to test if the exposure to higher doses of rhG-CSF for a France) reagents as previously described.14 brief time could yield enough progenitor cells to ensure engraftment. A target number for collection was 4 ϫ 106 CD34ϩ cells/kg b.w., and the apheresis was done at a blood ϩ Ͼ ϫ 6 CD34 cell count of 20 10 /l. The results in this group CFU-GEMM, BFU-E and CFU-GM of donors (group 3) were compared to the results in a group of donors with a standard and well described schema. The assay for progenitor cells was carried out as already described.15 Briefly, 1 ϫ 105 mononuclear cells were plated Evaluation in 35 mm Petri dishes in 1 ml aliquots of IMDM, containing 30% FCS (HCC4434; Stem Cell Technologies, Vancouver, Peripheral blood counts, whole blood counts and platelet Canada). Progenitor cell growth was evaluated after 14 counts were performed before and after apheresis using a days of incubation in a humidified atmosphere. Two dishes haematocytometer (Micro Diff II, Coulter, Krefeld, were set up for each individual data point per experiment. Germany). An aliquot of WBC was taken and used to cal- CFU-GEMM, BFU-E and CFU-GM were scored from the culate the CD34ϩ cell count. same dish according to previously published criteria.15

Bone Marrow Transplantation Dose of lenograstim for mobilization of PBSC N Basara et al 373 Statistical analysis CD34ϩ cells was also calculated per kg body weight (b.w.) of recipient, per kg b.w. of donor per apheresis and as When analysing the relationship between G-CSF dose and ϩ yield/blood volume processed. The differences between the blood CD34 levels, only values obtained on the day of ϩ three groups regarding CD34 cells per kg b.w. were also the first apheresis were used to avoid any influence of prior ϩ ± not significant. In addition to CD34 cells, the total number leukaphereses. The baseline numbers are shown as mean of CD3ϩ and CD16ϩ,56ϩ cells was determined. There was s.d. Multigroup comparison was performed by analysis of no significant difference in the total number of T cells variance (ANOVA).16 Differences between the mean values between any of the three groups of donors. were evaluated by paired t-test.16 To estimate correlation, Spearman’s correlation coefficient (R) was calculated. All the statistical analyses were performed using the Excel pro- Correlation between CD34ϩ cells in blood and the gram for windows 95. leukapheresis product The number of CD34ϩ cells in the blood on the day of Results apheresis was significantly correlated with the number of CD34ϩ cells in the apheresis product in groups 1, 2 and 3 = Ͻ = Ͻ = Since the optimal concentration for the mobilization of (r 0.68; P 0.001; r 0.78; P 0.001; and r 0.60; ϩ P Ͻ 0.001, respectively) (Figure 1), as well as in the whole CD34 cells using rhG-CSF and also the time frame for = Ͻ harvesting a sufficient number of CD34ϩ cells remain to group of donors (r 0.80; P 0.0001). be determined in allogeneic transplantation, we conducted an open label randomized split dose escalating study in a Correlation between donor characteristics and CD34ϩ total of 36 volunteer donors randomised in three groups yield receiving either 8, 11 or 15 ␮g/kg/day in split doses over ϩ a period of 4 to 5 days. WBC count increased in all three The number of CD34 cells pre-apheresis was inversely groups of donors to a level of eight- to nine-fold (Table correlated with age in donor groups 1 and 2 (r = 0.46; r = 1). When compared to the pre-treatment values, a slight 0.33; respectively) (Figure 2). In group 3, however, the ϩ reduction in the platelet count of 20% was noted in four number of CD34 cells pre-apheresis was not correlated patients. Apheresis was performed in groups 1 and 2 on with age. day 5 of mobilization in order to obtain a sufficient number of stem cells for allogeneic transplantation. In contrast, har- Circulating progenitor cells vest could be routinely performed on day 4 of stimulation with the split dose of 15 ␮g/kg/day. The total number of CFU-GM, BFU-E and CFU-GEMM were mobilized in per- WBC obtained by one apheresis was 4.83, 4.97 and ipheral blood at median numbers of 49.7, 59.2 and 4.67 ϫ 1010 in groups 1, 2 and 3, respectively. The total 21.4 ϫ 104/kg b.w. of recipient, BFU-E 114, 61, number of CD34ϩ cells harvested was 4.29, 3.34 and 46 ϫ 104/kg b.w. and CFU-GEMM at median numbers of 3.13 ϫ 108 in groups 1, 2 and 3, respectively. The differ- 7.1, 3.7 and 2 ϫ 104/kg b.w. of recipient. There was no ences in the total number of CD34ϩ cells between the three significant difference between the number of progenitor groups were not significant (Table 2). The total number of cells mobilized in the three groups of donors (Table 2).

Table 2 Total No. of WBC, CD34+, T, NK and CFC by one leukapheresis

Dose of lenograstim (split dose)

Group 1 Group 2 Group 3 (8 ␮g/kg/day) (11 ␮g/kg/day) (15 ␮g/kg/day)

No. of donors 10 14 12 Total WBC × 1010 4.83 ± 1.91 4.97 ± 1.90 4.67 ± 2.08 Total CD34+ × 108 4.29 ± 2.16 3.34 ± 1.82 3.13 ± 2.95 CD34 × 106/kg b.w. of recipient 6.19 ± 2.89 4.70 ± 2.61 4.12 ± 3.46 CD34 × 106/kg b.w. of donor per apheresis 5.30 ± 2.83 4.42 ± 2.27 3.86 ± 2.86 CD34 × 106 cells/l of blood processed 29.0 ± 13.4 25.6 ± 11.7 24.4 ± 26.5 Total CD3+ × 1010 1.60 ± 0.63 1.27 ± 0.71 1.54 ± 0.62 CD3 × 108/kg b.w. of recipient 2.34 ± 0.97 1.89 ± 1.33 2.10 ± 0.86 Total CD16+,56+ × 108 18.2 ± 9.28 16.5 ± 7.11 18.9 ± 10.4 CD16+,56+ × 106/kg b.w. of recipient 26.4 ± 12.2 23.7 ± 12.1 26.0 ± 16.1 CFC × 104/kg b.w. of recipient CFU-GM 49.7 (18–205) 59.2 (15–105) 21.4 (0.8–74.3) BFU-E 114 (38.7–268) 61.0 (7.1–371) 46.2 (6.5–258) CFU-GEMM 7.07 (1.04–57.8) 3.72 (1.45–12.8) 1.95 (0.78–133)

Values are expressed as mean ± s.d. Number of CFC is expressed as median (range).

Bone Marrow Transplantation Dose of lenograstim for mobilization of PBSC N Basara et al 374 aab 120 120 100 100 80 80 60 60 Yield Yield 40 40 20 20

0 0 0 50 100150 200 0 50 100150 200 CD34+ cells/µ l PB pre-apheresis CD34+ cells/µ l PB pre-apheresis

c d 120 120

100 100

80 80

60 60 Yield Yield 40 40

20 20 0 0 0 50 100150 200 250 050 100150 200 250 CD34+ cells/µ l PB pre-apheresis CD34+ cells/µ l PB pre-apheresis Figure 1 Correlation between CD34+ cells in blood and leukapheresis product. The efficiency of mobilization and leukapheresis was determined by calculating the number of CD34+ in the final component per volume of processed blood. Expression in this way compensates for individual differences in blood volume, number of total blood volumes processed and duration of leukapheresis. (a) Group 1, Pearson r = 0.772 (P Ͻ 0.008); (b) group 2, Pearsson r = 0.7067 (P Ͻ 0.0047); (c) group 3, Pearson r = 0.9432 (P Ͻ 0.0001); (d) all three groups, Pearson r = 0.8026 (P Ͻ 0.0001).

a b 250 250

200 200

150 150

100 100 50 50

CD34 pre-apheresis CD34 pre-apheresis 0 0 08020 40 60 0 10 20 30 40 50 Age Age

c d 250 250

200 200

150 150

100 100

50 50 CD34 pre-apheresis CD34 pre-apheresis 0 0 0 20 40 60 80 08020 40 60 Age Age

Figure 2 Correlation between CD34+ in blood and age. (a) Group 1, Pearson r = 0.4587; (b) group 2, Pearson r = 0.3309; (c) group 3, Pearson r = 0.1979; (d) all three groups, Pearson r = 0.4587 (P Ͻ 0.05).

Bone Marrow Transplantation Dose of lenograstim for mobilization of PBSC N Basara et al 375 Notably BFU-E were mobilized to a larger extent than CSF was well tolerated and resulted in high numbers of CFU-GM in all dose groups. CD34ϩ cells in apheresis products in healthy donors. Recently, Arbona et al19 similarly demonstrated that mobil- ization with 6 or 8 ␮g/kg/12 h of filgrastim was also asso- Discussion ciated with collection of significantly more CD34ϩ cells in comparison with 10 ␮g/kg/24 h. Weaver et al20 recently The optimal dosage of rhG-CSF for stem cell (CD34ϩ cells) evaluated doses of filgrastim up to 40 ␮g/kg/day but in an mobilization in normal donors is not well defined. Although autologous setting in patients with breast cancer. In patients rhG-CSF doses up to 24 ␮g/kg/day have been given,5,11 with good marrow reserves, doses of filgrastim greater than experience with such a dose range remains limited. In this 10 ␮g/kg/day yielded a higher number of CD34ϩ cells and study, we performed a randomized open label dose-finding may be useful when high numbers of CD34ϩ cells are trial and analysed the quality of harvest product mobilized required. with three different doses of lenograstim aiming to evaluate Lenograstim appears to be well tolerated during short- if the dose of 15 ␮g/kg/day given in a split regimen would term administration and there do not appear to be long-term result in the same or improved numbers of CD34ϩ cells adverse effects. With regard to safety issues, G-CSF has harvested earlier, ie at day ϩ4 after stimulation. We used been administered after several years of follow-up21 in allo- lenograstim, since it has been shown in a comparative study geneic-related PBSCT. PBSCT from unrelated donors has that lenograstim at 10 ␮g/kg/day mobilized PBSC more been increasingly used in EBMT programs.22 efficiently than an identical dose of filgrastim.12 We were It should be noted that in our study a remarkable varia- able to demonstrate that there was a significant correlation bility in the response to lenograstim was observed. Several between circulating CD34ϩ cells on the day of harvest and investigators18,23,24 have already reported this for lenogras- the CD34ϩ cells in the apheresis products in all three tim as well as for filgrastim. It would be important to ident- groups. In addition to this, the mobilization of colony-for- ify donors with poor mobilization, in order to prime them ming cells was similar in all three groups, although a trend by different strategies, usually including combinations of for fewer progenitors in group 3 was observed. A possi- cytokines.25 bility that earlier collection results in fewer ‘useful’ pro- In conclusion, the mobilization of allogeneic PBPC is genitors could not be neglected. However, all of the patients feasible using short-term administration of different doses receiving PBSCT from group 3 of donors had sustained of lenograstim. The collection of allogeneic PBPC is safe engraftment, indicating that colony-forming cells are not at day 4 if split doses of 15 ␮g/kg/day are used for mobiliz- the only cells responsible for successful engraftment. The ation and could be effectively used also in older donors. most important finding of our study was that a similar num- ber of CD34ϩ cells in the apheresis product could be obtained with one apheresis by using a dose of 15 References ␮g/kg/day lenograstim for only 4 days. In addition, we showed that the number of CD34ϩ cells pre-apheresis was 1 Dreger P, Suttorp M, Haferlach T et al. Allogeneic granulo- not significantly inversely correlated with age, in the group cyte colony-stimulating factor-mobilized peripheral blood pro- ␮ genitor cells for treatment of engraftment failure after bone of donors mobilized with 15 g/kg/day. However, the lack marrow transplantation. Blood 1993; 81: 1404–1407. of this correlation could also be due to the small numbers 2 Russel NH, Hunter AE. Peripheral blood stem cells for allo- of patients and variability in this group. In particular, the geneic transplantation. Bone Marrow Transplant 1994; 13: incidence of side events in this group of donors was not 353–355. increased in comparison to the donors mobilized with lower 3 Bensinger WI, Weaver CH, Appelbaum FR et al. Transplan- doses of lenograstim. The dose of 15 ␮g/kg/day in a split tation of allogeneic peripheral blood stem cells mobilized by regimen might be considered for volunteer donors, who recombinant human granulocyte colony-stimulating factor. turn out to be poor mobilizers, such as elderly people. Blood 1995; 85: 1655–1658. With regard to the dosage used to harvest volunteer 4 Schmitz N, Bacigalupo A, Labopin M et al on behalf of the donors after lenograstim administration, a consensus has European Group for Blood and Marrow Transplantation (EBMT). Transplantation of peripheral blood progenitor cells not yet been achieved. There is evidence that doses of less ␮ from HLA-identical sibling donors. Br J Haematol 1996; 95: than 10 g/kg/day are less effective in increasing circulat- 715–723. ϩ 10 ing numbers of CD34 cells. Factors influencing yields 5 Bensinger WI, Clift R, Martin P et al. Allogeneic peripheral of progenitor cells for allogeneic transplantation have been blood stem cell transplantation in patients with advanced hem- recently discussed.17 Luider et al17 have suggested that a atologic malignancies: a retrospective comparison with mar- daily filgrastim dose of 7.5 to 10 ␮g/kg (intermediate dose) row transplantation. Blood 1996; 88: 2794–2800. is a convenient regimen giving adequate yields from a sin- 6 Schmitz N, Bacigalupo A, Hasenclever D et al. Allogeneic gle collection on day 4 or 5 in most donors. However, bone marrow transplantation vs filgrastim-mobilised periph- according to these authors, a collection on day 4 may also eral blood progenitor cell transplantation in patients with early be suitable for most donors mobilized with intermediate- leukaemia: first results of a randomised multicentre trial of the Ͼ ␮ European Group for Blood and Marrow Transplantation. Bone and high-dose ( 10 g/kg) filgrastim. Other groups have Marrow Transplant 1998; 21: 995–1003. reported series of normal donors, who have been mobilized 7 Russel NH, Hunter A, Roger S et al. Peripheral blood stem with rhG-CSF using amounts ranging from 2.5 to 24 cells as an alternative to marrow for allogeneic transplantation. ␮g/kg/day.10,11,18 Waller et al11 showed that a high-dose (24 Lancet 1993; 341: 1482–1483. ␮g/kg/day) twice daily regimen of nonglycosylated rhG- 8 Lane TA, Law P, Maruyama M et al. Harvesting and enrich-

Bone Marrow Transplantation Dose of lenograstim for mobilization of PBSC N Basara et al 376 ment of hematopoietic progenitor cells mobilized into the per- peripheral blood progenitor cells (PBPC) mobilized by filgra- ipheral blood of normal donors by granulocyte–macrophage stim (rHuG-CSF) in normal volunteers: dose effect relation- colony-stimulating factor (GM-CSF) or G-CSF: potential role ship for filgrastim with the character of mobilized PBPC. Br in allogeneic marrow transplantation. Blood 1995; 85: 275– J Haematol 1996; 92: 705–803. 282. 19 Arbona C, Prosper F, Benet I et al. Comparison between once 9 Goldman J. Peripheral blood stem cells for allografting. Blood a day vs twice a day G-CSF for mobilization of peripheral 1995; 85: 1413–1415. blood progenitor cells (PBPC) in normal donors for allogeneic 10 Ho¨glund M, Smedmyr B, Simonsson B et al. Dose-dependent PBPC transplantation. Bone Marrow Transplant 1998; 22: mobilisation of haematopoietic progenitor cells in healthy vol- 39–45. unteers receiving glycosylated rHuG-CSF. Bone Marrow 20 Weaver CH, Birch R, Greco FA et al. Mobilization and har- Transplant 1996; 18: 19–27. vesting of peripheral blood stem cells: randomized evaluations 11 Waller TF, Bertz H, Wenger MK et al. Mobilisation of periph- of different doses of filgrastim. Br J Haematol 1998; 100: eral blood progenitor cells for allogeneic transplantation: effi- 338–347. cacy and toxicity of a high-dose rhG-CSF regimen. Bone Mar- 21 Dreger P, Haferlach T, Eckstein V et al. G-CSF mobilized row Transplant 1996; 18: 279–283. peripheral blood progenitor cells for allogeneic transplan- 12 Ho¨glund M, Smedmyr B, Bengtsson M et al. Mobilization of tation: safety, kinetics of mobilization and composition of the CD34 cells by glycosylated and nonglycosylated G-CSF in graft. Br J Haematol 1994; 87: 609–613. healthy volunteers – a comparative study. Eur J Haematol 22 Ringden O, Remberger M, Runde V et al. Peripheral blood 1997; 59: 177–183. stem cell (PBSC) transplantation from unrelated donors: a 13 Anderlini P, Przepiorka D, Seong C et al. Factors affecting comparison with marrow transplantation. Blood 1999; 94: mobilization of CD34ϩ cells in normal donors treated with 455–464. filgrastim. Transfusion 1997; 37: 507–512. 23 Grigg AP, Roberts AW, Raunow H et al. Optimizing dose 14 Kreissig C, Kirsch A, Serke S. Characterization and measure- and scheduling of filgrastim (granulocyte colony-stimulating ment of CD34 expressing hematopoietic cells. J Chemother factor) for mobilisation and collection of peripheral blood pro- 1994; 3: 263–289. genitor cells in normal volunteers. Blood 1995; 86: 4437– 15 Fauser AA, Messner HA. Granuloerythropoietic colonies in 4445. human bone marrow, peripheral blood and cord blood. Blood 24 Weaver CH, Buckner CD, Longin K et al. Syngeneic trans- 1978; 52: 1243–1248. plantation with peripheral blood mononuclear cells collected 16 Steel RGD, Torrie JH. Principles and Procedures of Statistics. after administration of recombinant human granulocyte colony McGraw-Hill: New York, 1960. stimulating factor. Blood 1993; 82: 1981–1984. 17 Luider J, Brown C, Selinger S et al. Factors influencing yields 25 Geissler K, Peschel C, Niederwieser D et al. Potentiation of of progenitor cells for allogeneic transplantation: optimization granulocyte colony stimulating factor-induced mobilization of of G-CSF dose, day of collection and duration of leukapher- circulating progenitor cells by seven-day pretreatment with esis. J Hematother 1997; 6: 575–580. interleukin-3. Blood 1996; 87: 2732–2739. 18 Tanaka R, Matsudaira T, Aizawa J et al. Characterization of

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