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Large volume leukapheresis with AMICUS cell separator in peripheral blood stem cell autologous transplant
Transfusion and Apheresis Science 24 (2001) 79±83
www.elsevier.com/locate/transci
Large volume leukapheresis with AMICUS cell separator in peripheral blood stem cell autologous transplant Patrizia Accorsi a, Mario Dell'Isola a, Tiziana Bon®ni a, Raaella Giancola a, Antonio Spadano b, Giuseppe Fioritoni b, Antonio Iacone a,* a
Department of Transfusion Medicine, Centro Studi `E. Jucci' Ciancarelli, Ospedale Civile, Pescara, Italy b Department of Haematology, Ospedale Civile, Pescara, Italy
1. Introduction The use of peripheral blood stem cell (PBSC) collection has dramatically increased since the ®rst human PBSC transplantation (PBSCT) reported in 1979 and it has now become a widely accepted therapeutic option for patients with chemotherapy-sensitive malignancies. PBSCT oers some advantages over bone marrow transplantation: 1. more rapid granulocyte and platelet engraftment, 2. reduction of supportive care and costs, 3. PBSC collection without general anesthesia. These advantages are largely derived from the very high number of hemopoietic stem cells circulating in the patient's blood stream after mobilization with chemotherapy and/or cytokine administration [1,2]. The collection of a target CD34+ dose depends on: 1. biological variables (precount of CD34+), 2. device eciency, 3. procedure parameters such as total whole blood processed (WBP). Previous experience has proven the usefulness in extending the apheresis procedure to large volume leukapheresis (LVL) that is de®ned by pro-
*
Corresponding author.
cessing more than three times the total WBP. LVL permits one to reach a higher dose of CD34+ than the standard procedure due to both the volume of blood processed and the continuous intraprocedure CD34+ cell recruitment as demonstrated in patients and donors [3±7]. The aim of this study is to evaluate a new software program, version 2.42 for MNC collection with the AMICUS cell separator and its performance in hemato-oncology patients. In particular, we evaluated the LVL impact on the patient's safety, the characteristics of the products after 5, 10 and 15 l of WBP, the suitability of the product for immunoselection and its clinical ecacy.
2. Materials and methods 2.1. Device The AMICUS cell separator permits the intermittent collection of mononuclear cells (MNC) by using a continuous blood ¯ow. As a matter of fact, the MNC are recirculated and concentrated in the single chamber of the belt with a periodic transfer to the storage bag located outside the centrifuge. The transfer depends on the blood volume processed per cycle. In every procedure, the following parameters
1473-0502/01/$ - see front matter Ó 2001 Elsevier Science Ltd. All rights reserved. PII: S 0 9 5 5 - 3 8 8 6 ( 0 0 ) 0 0 1 3 0 - 2
80
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83
were set: donor's weight, RBC oset volume, WBP volume per cycle, total number of cycles per procedure. The rate of anticoagulant±citrate±dextrose formula A (ACD-A, Baxter) was 1.5 mg/kg per min (12:1 ratio of blood to ACD). Calcium gluconate (50 mg) in 500 ml of saline solution was infused through the return-line injection site during the procedure and the patients were monitored for any adverse reactions (i.e., citrate toxicity). 2.2. Patients We enrolled 12 patients (4 males, 8 females) diagnosed as follows: 5 non-Hodgkin's lymphoma (nHL), 2 solid tumors (ST), 2 multiple myeloma (MM), 1 acute lymphoblastic leukemia (ALL), 1 acute myeloid leukemia (AML) and 1 Hodgkin's disease (HD). The median age was 48.5 years (range 24±60) and weight in kilograms was 68.5 (range 45±88). In 10 patients the target dose of CD34+/kg was 5 106 even after immunoselection, in the 2 MM patients the target dose was doubled because they were scheduled to undergo a tandem transplant. Fifteen out of 20 PBSC products were immunoselected with Isolex 300i (Nexell) by a method largely employed in an allogeneic setting [8]. In nHL and MM products a combined immunoselection with monoclonal antibody cocktail anti-B (Anti CD19-22, Anti CD20 and Anti CD23 Nexell) were performed as well as in ALL (Anti CD10, Anti CD19 and Anti CD20). In ovarian cancer, a positive selection was performed. The suitability to submit the product to immunoselection was evaluated by measuring the percentage of ®nal CD34+ recovery and purity. The PBSC/CD34+ were frozen in a controlled rate freezer and stored in a mechanical freezer (temperature 135°C) according to our standard operating procedure. The speed of engraftment (days to reach 0:5 109 =l granulocytes (PMN) and 20 109 =l platelets (PLT) and the overall survival were considered as criteria to evaluate the clinical ecacy of these products.
procedure at 5, 10 and 15 l of WBP and ®nal and partial yields. The collection bag was emptied and the product transferred in the second collection bag; therefore, bag 1 (5 l), bag 2 (10 l) and bag 3 (15 l) of the partial products were evaluated separately.The samples were counted with an automated cell counter (Sysmex SE 9000, Dasit). The MNC fraction was evaluated with a ¯ow-cytometry analysis using double staining CD45(HLe-1) FITC/CD14 (LeuM3)PE. Cell viability was assessed by 7-aminoactinomycin D (7-AAD). The cyto¯uorimetric analysis was assessed with a FACScane ¯ow cytometer (Becton Dickinson immunocytometry system), equipped with the Cell Queste software. The International Society for Hematotherapy and Graft Engineering (ISHAGE) guidelines for CD34+ stem cell analysis were followed. PBSC products and peripheral blood were sampled at the start and the end of leukapheresis assessed for colony forming units (CFUs). Clonogenic assays were performed in duplicate using a commercial methylcellulose-based medium supplemented with a recombinant human stem cell factor, interleukin-3, granulocyte-macrophagecolony stimulating factor and erythropoietin (GF 4444, Stem Cell Technologies, Vancouver, BC, Canada). The partial and total collection eciencies for MNC and CD34+ cells were calculated by dividing the number of cells in the product by the number of cells processed during the apheresis procedure. The processed cells were calculated using the following formula: pre + post peripheral blood (MNC or CD34+) cell count=2 WBP ml anticoagulant volume ml. 2.4. Statistical analysis The results were given as median and range. The dierences and statistical signi®cance of the partial products were determined using the Kruskal±Wallis test. A signi®cance level of P < 0:05 was chosen.
2.3. Cell count and analysis
3. Results
MNC and CD34+ cell counts of the peripheral blood were scheduled as follows: pre and post
From May to August 1999, 20 LVL were performed. On the 1st day of collection the patient's
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83
median peripheral precount values of WBC 109 =l, MNC 109 =l, PLT 109 =l and CD34=ll were 9.46 (2.62±45.3), 1.78 (0.7±7.1), 106 (34±332) and 36 (12±567), respectively. Apheresis procedures were performed on a daily basis, 8 patients underwent 2 procedures and 4 patients 1 procedure. A median of 3.1 (2.8±4.1) times the total blood volume per donor (WBP/BV) were processed. Details of procedure parameters are summarized in Table 1. Apheresis related adverse events were restricted to a mild citrate toxicity (circumoral and limb paresthesias). The symptoms were resolved with an addition of 20 mgr calcium gluconate infusion. A volume of 303 (242±347) ml of PBSC enriched product contained a median of 73.7% (45.1±96.3) of MNC, 1.6% (0.1±7.7) of CD34+ cells and the cell viability was 99% (97.5±99.7). The apheresis collections had a total median
81
of 23:8 6:1±49:3 109 , 17:5 3:5±40:9 109 , 324 69±3801 106 and 1872 308±20567 104 of WBC, MNC, CD34+ and CFU-GM, respectively. The median MNC, CD34+ and CFU-GM/ kg recipient's weight per procedure were 2:64 0:4±5:5 108 , 5 1:2±43 106 and 28 4±234 104 , respectively. The collection eciencies of MNC and CD34+ cells were 61.9% (22.3±102) and 63.9% (29±111), respectively. During the run, a continuous increase in MNC and CD34+ values and a signi®cative increment (P 0:02 and P 0:004) of MNC and CD34+ eciencies, respectively, were observed (Table 2). The behaviour of PLTs and the platelet drop did not show any signi®cative dierence (P 0:5) during the entire procedure (Table 3). Another aspect of LVL relates to the contamination of unwanted cells in the ®nal product. The ®nal products contained a PMN concentration of
Table 1 Procedure parameters Whole blood per cycle (ml) RBC oset volume Flow rate (ml/min) Whole blood processed (ml) ACD (ml) Number of cycles Time (min)
1000 (2 procedures) 6.5 (16 procedures) 68 (47±80) 15963 (12 623±17 657) 1504 (1152±1646) 13 (10±13) 306 (275±346)
1400 (18 procedures) 7 (4 procedures)
Table 2 Results of partial products and collection eciencies for 0±5, 5±10, 10±15 l of WBP and the statistical analysis WBP (l) WBP/BV (times) MNC 109 CD34 106 MNC eciency (%) CD34+ eciency (%) *
0±5 0.9 (0.7±1.4) 4.5 (0.2±7.2) 93 (12±541) 45 (23±100) 44 (24±95)
5±10 0.9 (0.7±1.4) 5.0 (0.8±8.1) 104 (13±659) 69 (25±112) 83 (22±148)
p ns ns ns 0.02 0.004
10±15 0.8 (0.7±1.0) 5.1 (1.3±7.3) 112 (41±596) 72 (24±131) 80 (34±146)
Kruskal±Wallis.
Table 3 Behaviour of platelets during the procedures WBP (l) PLT precount 109 =l PLT postcount 109 =l Drop PLT % *
Kruskal±Wallis.
0±5 89.5 (34±332) 88.5 (30±282) 5 (0±25)
5±10 88.5 (30±282) 79.5 (27±258) 9.7 (0±28)
10±15 63 (36±258) 60 (33±245) 7.5 (0±30)
p ns ns ns
82
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83
Table 4 Transplantation data #Pat.
04.99 05.99 07.99 09.99 10.99 10.99a a
Disease
nHL nHL AML Ovarian C MM MM
CD34+ infused (106 =kg) 5.5 5.3 8.5 6.8 5.1 5.1
Days of take
Transfusion support
PMN > 0:5 109 =l
PLT > 20 109 =l
T GR
T PLT
Hospitalization day
12 11 8 10 11 10
15 15 22 10 12 12
2 4 2 0 2 2
2 3 4 2 4 4
22 21 25 21 26 17
Second transplantation.
26% (3.7±54.9), PLT contamination of 1:3 0:3± 4:1 1011 and a volume of red blood cell (RBC) equal to 16 ml (5.6±48). In the 15 PBSC products submitted to the immunoselection, the recovery and purity of CD34+ was 66% (48±81) and 96% (89±98.5), respectively. 3.1. Transplant results Five out of 12 patients were submitted to a transplantation and 1 patient with MM underwent transplantation twice. The patient with acute leukemia was grafted with unmanipulated PBSC, while the other patients were grafted with immunoselected cells (Table 4). Up to now, all patients are alive and in complete remission.
4. Discussion In this study we focused our attention towards the performance of the new blood cell separator AMICUS±Baxter in an autologous PBSC collection using a LVL procedure. This device equipped with the software version 2.42 is safe and eective. No serious adverse donor reactions attributable to the device were registered and no procedure was interrupted. The only negative aspect observed in these procedures was the lengthy procedure time. In 11/20 cases with a single apheresis the target dose of CD34 P5 106 =kg was reached. Only in 3 procedures the harvested dose was less than 2:7 106 =kg. In 14/20 procedures, the ®nal PLT contamination in the PBSC products was less than
1:9 1011 . We had no technical problems regarding the quality of graft during the immunoselection with Isolex 300i. ``In vitro'' results in terms of recovery and purity are comparable to the ones achieved with cells collected by CS3000 (data unpublished). Despite the LVL, the platelet postcount was always higher than 20 109 =l without the need of transfusion support. Our data strongly supports the usefulness of LVL with AMICUS in collecting higher MNC and CD34+ doses in one or two apheresis and in improving the collection eciency. This latter shows a favourable trend during the entire procedure both for MNC and CD34+. All transplanted patients promptly engrafted and are currently alive and well. The speed of engraftment, transfusional support and hospitalization were perfectly comparable with our previous historical data.
References [1] Gray TF, Shea TC. Current status of peripheral blood progenitor cell transplantation. Semin Oncol 1994;21:93± 101. [2] Lee JH, Klein HG. Collection and use of circulating hematopoietic progenitor cells. Hematol Oncol Clin North Am 1995;9:1±22. [3] Weaver CH, Hazelton B, Birch R et al. An analysis of engraftment kinetics as a function of the CD34 content of peripheral blood progenitor cell collections in 692 patients after the administration of myeloablative chemotherapy. Blood 1995;86:3961±9. [4] Malachowski ME, Comenzo RL, Hillyer CD et al. Largevolume leukapheresis for peripheral blood stem cell collec-
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83 tion in patients with hematologic malignancies. Transfusion 1992;32:729±31. [5] Accorsi P, Dell'Isola M, Bon®ni T et al. Large volume leukapheresis (LVL) in PBSC collection using AMICUSFENWAL blood cell separator. Bone Marrow Transplant 2000;25S1:S267. [6] Humpe A, Riggert J, Munzel U et al. A prospective, randomized, sequential crossover trial of large-volume leukapheresis and normal-volume leukaphereses: eect on
83
progenitor cells and engraftment. Transfusion 1999; 39:1120±7. [7] Bon®ni T, Accorsi P, Giancola R et al. Kinetic of intraprocedure CD34+ cell release during PBSC collection in healthy donors. Exp Hematol 2000;28S1(7):117. [8] Accorsi P, Bon®ni T, Dell'Isola M. PBSC immunoselection with Isolex 300i in allogeneuic transplantation. Blood 1999;94(10):S617.
www.elsevier.com/locate/transci
Large volume leukapheresis with AMICUS cell separator in peripheral blood stem cell autologous transplant Patrizia Accorsi a, Mario Dell'Isola a, Tiziana Bon®ni a, Raaella Giancola a, Antonio Spadano b, Giuseppe Fioritoni b, Antonio Iacone a,* a
Department of Transfusion Medicine, Centro Studi `E. Jucci' Ciancarelli, Ospedale Civile, Pescara, Italy b Department of Haematology, Ospedale Civile, Pescara, Italy
1. Introduction The use of peripheral blood stem cell (PBSC) collection has dramatically increased since the ®rst human PBSC transplantation (PBSCT) reported in 1979 and it has now become a widely accepted therapeutic option for patients with chemotherapy-sensitive malignancies. PBSCT oers some advantages over bone marrow transplantation: 1. more rapid granulocyte and platelet engraftment, 2. reduction of supportive care and costs, 3. PBSC collection without general anesthesia. These advantages are largely derived from the very high number of hemopoietic stem cells circulating in the patient's blood stream after mobilization with chemotherapy and/or cytokine administration [1,2]. The collection of a target CD34+ dose depends on: 1. biological variables (precount of CD34+), 2. device eciency, 3. procedure parameters such as total whole blood processed (WBP). Previous experience has proven the usefulness in extending the apheresis procedure to large volume leukapheresis (LVL) that is de®ned by pro-
*
Corresponding author.
cessing more than three times the total WBP. LVL permits one to reach a higher dose of CD34+ than the standard procedure due to both the volume of blood processed and the continuous intraprocedure CD34+ cell recruitment as demonstrated in patients and donors [3±7]. The aim of this study is to evaluate a new software program, version 2.42 for MNC collection with the AMICUS cell separator and its performance in hemato-oncology patients. In particular, we evaluated the LVL impact on the patient's safety, the characteristics of the products after 5, 10 and 15 l of WBP, the suitability of the product for immunoselection and its clinical ecacy.
2. Materials and methods 2.1. Device The AMICUS cell separator permits the intermittent collection of mononuclear cells (MNC) by using a continuous blood ¯ow. As a matter of fact, the MNC are recirculated and concentrated in the single chamber of the belt with a periodic transfer to the storage bag located outside the centrifuge. The transfer depends on the blood volume processed per cycle. In every procedure, the following parameters
1473-0502/01/$ - see front matter Ó 2001 Elsevier Science Ltd. All rights reserved. PII: S 0 9 5 5 - 3 8 8 6 ( 0 0 ) 0 0 1 3 0 - 2
80
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83
were set: donor's weight, RBC oset volume, WBP volume per cycle, total number of cycles per procedure. The rate of anticoagulant±citrate±dextrose formula A (ACD-A, Baxter) was 1.5 mg/kg per min (12:1 ratio of blood to ACD). Calcium gluconate (50 mg) in 500 ml of saline solution was infused through the return-line injection site during the procedure and the patients were monitored for any adverse reactions (i.e., citrate toxicity). 2.2. Patients We enrolled 12 patients (4 males, 8 females) diagnosed as follows: 5 non-Hodgkin's lymphoma (nHL), 2 solid tumors (ST), 2 multiple myeloma (MM), 1 acute lymphoblastic leukemia (ALL), 1 acute myeloid leukemia (AML) and 1 Hodgkin's disease (HD). The median age was 48.5 years (range 24±60) and weight in kilograms was 68.5 (range 45±88). In 10 patients the target dose of CD34+/kg was 5 106 even after immunoselection, in the 2 MM patients the target dose was doubled because they were scheduled to undergo a tandem transplant. Fifteen out of 20 PBSC products were immunoselected with Isolex 300i (Nexell) by a method largely employed in an allogeneic setting [8]. In nHL and MM products a combined immunoselection with monoclonal antibody cocktail anti-B (Anti CD19-22, Anti CD20 and Anti CD23 Nexell) were performed as well as in ALL (Anti CD10, Anti CD19 and Anti CD20). In ovarian cancer, a positive selection was performed. The suitability to submit the product to immunoselection was evaluated by measuring the percentage of ®nal CD34+ recovery and purity. The PBSC/CD34+ were frozen in a controlled rate freezer and stored in a mechanical freezer (temperature 135°C) according to our standard operating procedure. The speed of engraftment (days to reach 0:5 109 =l granulocytes (PMN) and 20 109 =l platelets (PLT) and the overall survival were considered as criteria to evaluate the clinical ecacy of these products.
procedure at 5, 10 and 15 l of WBP and ®nal and partial yields. The collection bag was emptied and the product transferred in the second collection bag; therefore, bag 1 (5 l), bag 2 (10 l) and bag 3 (15 l) of the partial products were evaluated separately.The samples were counted with an automated cell counter (Sysmex SE 9000, Dasit). The MNC fraction was evaluated with a ¯ow-cytometry analysis using double staining CD45(HLe-1) FITC/CD14 (LeuM3)PE. Cell viability was assessed by 7-aminoactinomycin D (7-AAD). The cyto¯uorimetric analysis was assessed with a FACScane ¯ow cytometer (Becton Dickinson immunocytometry system), equipped with the Cell Queste software. The International Society for Hematotherapy and Graft Engineering (ISHAGE) guidelines for CD34+ stem cell analysis were followed. PBSC products and peripheral blood were sampled at the start and the end of leukapheresis assessed for colony forming units (CFUs). Clonogenic assays were performed in duplicate using a commercial methylcellulose-based medium supplemented with a recombinant human stem cell factor, interleukin-3, granulocyte-macrophagecolony stimulating factor and erythropoietin (GF 4444, Stem Cell Technologies, Vancouver, BC, Canada). The partial and total collection eciencies for MNC and CD34+ cells were calculated by dividing the number of cells in the product by the number of cells processed during the apheresis procedure. The processed cells were calculated using the following formula: pre + post peripheral blood (MNC or CD34+) cell count=2 WBP ml anticoagulant volume ml. 2.4. Statistical analysis The results were given as median and range. The dierences and statistical signi®cance of the partial products were determined using the Kruskal±Wallis test. A signi®cance level of P < 0:05 was chosen.
2.3. Cell count and analysis
3. Results
MNC and CD34+ cell counts of the peripheral blood were scheduled as follows: pre and post
From May to August 1999, 20 LVL were performed. On the 1st day of collection the patient's
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83
median peripheral precount values of WBC 109 =l, MNC 109 =l, PLT 109 =l and CD34=ll were 9.46 (2.62±45.3), 1.78 (0.7±7.1), 106 (34±332) and 36 (12±567), respectively. Apheresis procedures were performed on a daily basis, 8 patients underwent 2 procedures and 4 patients 1 procedure. A median of 3.1 (2.8±4.1) times the total blood volume per donor (WBP/BV) were processed. Details of procedure parameters are summarized in Table 1. Apheresis related adverse events were restricted to a mild citrate toxicity (circumoral and limb paresthesias). The symptoms were resolved with an addition of 20 mgr calcium gluconate infusion. A volume of 303 (242±347) ml of PBSC enriched product contained a median of 73.7% (45.1±96.3) of MNC, 1.6% (0.1±7.7) of CD34+ cells and the cell viability was 99% (97.5±99.7). The apheresis collections had a total median
81
of 23:8 6:1±49:3 109 , 17:5 3:5±40:9 109 , 324 69±3801 106 and 1872 308±20567 104 of WBC, MNC, CD34+ and CFU-GM, respectively. The median MNC, CD34+ and CFU-GM/ kg recipient's weight per procedure were 2:64 0:4±5:5 108 , 5 1:2±43 106 and 28 4±234 104 , respectively. The collection eciencies of MNC and CD34+ cells were 61.9% (22.3±102) and 63.9% (29±111), respectively. During the run, a continuous increase in MNC and CD34+ values and a signi®cative increment (P 0:02 and P 0:004) of MNC and CD34+ eciencies, respectively, were observed (Table 2). The behaviour of PLTs and the platelet drop did not show any signi®cative dierence (P 0:5) during the entire procedure (Table 3). Another aspect of LVL relates to the contamination of unwanted cells in the ®nal product. The ®nal products contained a PMN concentration of
Table 1 Procedure parameters Whole blood per cycle (ml) RBC oset volume Flow rate (ml/min) Whole blood processed (ml) ACD (ml) Number of cycles Time (min)
1000 (2 procedures) 6.5 (16 procedures) 68 (47±80) 15963 (12 623±17 657) 1504 (1152±1646) 13 (10±13) 306 (275±346)
1400 (18 procedures) 7 (4 procedures)
Table 2 Results of partial products and collection eciencies for 0±5, 5±10, 10±15 l of WBP and the statistical analysis WBP (l) WBP/BV (times) MNC 109 CD34 106 MNC eciency (%) CD34+ eciency (%) *
0±5 0.9 (0.7±1.4) 4.5 (0.2±7.2) 93 (12±541) 45 (23±100) 44 (24±95)
5±10 0.9 (0.7±1.4) 5.0 (0.8±8.1) 104 (13±659) 69 (25±112) 83 (22±148)
p ns ns ns 0.02 0.004
10±15 0.8 (0.7±1.0) 5.1 (1.3±7.3) 112 (41±596) 72 (24±131) 80 (34±146)
Kruskal±Wallis.
Table 3 Behaviour of platelets during the procedures WBP (l) PLT precount 109 =l PLT postcount 109 =l Drop PLT % *
Kruskal±Wallis.
0±5 89.5 (34±332) 88.5 (30±282) 5 (0±25)
5±10 88.5 (30±282) 79.5 (27±258) 9.7 (0±28)
10±15 63 (36±258) 60 (33±245) 7.5 (0±30)
p ns ns ns
82
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83
Table 4 Transplantation data #Pat.
04.99 05.99 07.99 09.99 10.99 10.99a a
Disease
nHL nHL AML Ovarian C MM MM
CD34+ infused (106 =kg) 5.5 5.3 8.5 6.8 5.1 5.1
Days of take
Transfusion support
PMN > 0:5 109 =l
PLT > 20 109 =l
T GR
T PLT
Hospitalization day
12 11 8 10 11 10
15 15 22 10 12 12
2 4 2 0 2 2
2 3 4 2 4 4
22 21 25 21 26 17
Second transplantation.
26% (3.7±54.9), PLT contamination of 1:3 0:3± 4:1 1011 and a volume of red blood cell (RBC) equal to 16 ml (5.6±48). In the 15 PBSC products submitted to the immunoselection, the recovery and purity of CD34+ was 66% (48±81) and 96% (89±98.5), respectively. 3.1. Transplant results Five out of 12 patients were submitted to a transplantation and 1 patient with MM underwent transplantation twice. The patient with acute leukemia was grafted with unmanipulated PBSC, while the other patients were grafted with immunoselected cells (Table 4). Up to now, all patients are alive and in complete remission.
4. Discussion In this study we focused our attention towards the performance of the new blood cell separator AMICUS±Baxter in an autologous PBSC collection using a LVL procedure. This device equipped with the software version 2.42 is safe and eective. No serious adverse donor reactions attributable to the device were registered and no procedure was interrupted. The only negative aspect observed in these procedures was the lengthy procedure time. In 11/20 cases with a single apheresis the target dose of CD34 P5 106 =kg was reached. Only in 3 procedures the harvested dose was less than 2:7 106 =kg. In 14/20 procedures, the ®nal PLT contamination in the PBSC products was less than
1:9 1011 . We had no technical problems regarding the quality of graft during the immunoselection with Isolex 300i. ``In vitro'' results in terms of recovery and purity are comparable to the ones achieved with cells collected by CS3000 (data unpublished). Despite the LVL, the platelet postcount was always higher than 20 109 =l without the need of transfusion support. Our data strongly supports the usefulness of LVL with AMICUS in collecting higher MNC and CD34+ doses in one or two apheresis and in improving the collection eciency. This latter shows a favourable trend during the entire procedure both for MNC and CD34+. All transplanted patients promptly engrafted and are currently alive and well. The speed of engraftment, transfusional support and hospitalization were perfectly comparable with our previous historical data.
References [1] Gray TF, Shea TC. Current status of peripheral blood progenitor cell transplantation. Semin Oncol 1994;21:93± 101. [2] Lee JH, Klein HG. Collection and use of circulating hematopoietic progenitor cells. Hematol Oncol Clin North Am 1995;9:1±22. [3] Weaver CH, Hazelton B, Birch R et al. An analysis of engraftment kinetics as a function of the CD34 content of peripheral blood progenitor cell collections in 692 patients after the administration of myeloablative chemotherapy. Blood 1995;86:3961±9. [4] Malachowski ME, Comenzo RL, Hillyer CD et al. Largevolume leukapheresis for peripheral blood stem cell collec-
P. Accorsi et al. / Transfusion and Apheresis Science 24 (2001) 79±83 tion in patients with hematologic malignancies. Transfusion 1992;32:729±31. [5] Accorsi P, Dell'Isola M, Bon®ni T et al. Large volume leukapheresis (LVL) in PBSC collection using AMICUSFENWAL blood cell separator. Bone Marrow Transplant 2000;25S1:S267. [6] Humpe A, Riggert J, Munzel U et al. A prospective, randomized, sequential crossover trial of large-volume leukapheresis and normal-volume leukaphereses: eect on
83
progenitor cells and engraftment. Transfusion 1999; 39:1120±7. [7] Bon®ni T, Accorsi P, Giancola R et al. Kinetic of intraprocedure CD34+ cell release during PBSC collection in healthy donors. Exp Hematol 2000;28S1(7):117. [8] Accorsi P, Bon®ni T, Dell'Isola M. PBSC immunoselection with Isolex 300i in allogeneuic transplantation. Blood 1999;94(10):S617.