Peripheral Blood Stem Cell Collection Research Articles

The use of hematocrit level for predicting the efficiency of peripheral blood CD34(+) cell collection after G-CSF Mobilization in Healthy Donors.

Recently, peripheral blood stem cell (PBSC) has been widely used and replaced bone marrow (BM) as the stem cell source in allogeneic hematopoietic stem cell transplantation (HSCT) because of a more rapid engraftment, easier accessibility, and lower risk of donor complications. We, therefore, report the predicting factors for the high PBSC harvest yields in 50 healthy donors. Among the 50 donors, median collected CD34(+) cell number was 4.6 × 10(6/) kg (1.5-16.3 × 10(6) /kg). Number of circulating CD34+ cells and hematocrit (HCT) level increased parallelly whereas peripheral CD34+ cell numbers were decreased with increasing donor age. In univariate analysis, HCT level≥ 35.5% at the time of PBSC collection was significantly associated with high PBSC number (≥ 5.0 × 10(6) cells/kg) and donor aged <30 years was significantly associated with collected CD34+ cells ≥ 6.0 × 10(6) /kg, P = 0.03. HCT level ≥35.5% was an independent parameter for high WBC count (≥50 × 10(9) /L), P < 0.05. None of donor who had both HCT < 35.5% and WBC < 50 × 10(9) /L had circulating CD34+ cells ≥ 5.0 × 10(6) /kg. Platelet count ≥ 200 × 10(9) /L was found significantly in donors with WBC ≥ 40 × 10(9) /L (P = 0.03) and HCT ≥ 35.5%, P < 0.05. Collected PBSC number tended to be higher in our donors with high levels of HCT, WBC, and platelet. We also found that HCT and platelet levels in our donors decreased after receiving G-CSF administration compared with the initial complete blood counts (CBC) results. We, therefore, concluded that HCT level at the time of initiation leukapheresis was an important predictor for PBSC collection yields.

Tbo-Filgrastim Versus Filgrastim during Mobilization and Neutrophil Engraftment in Autologous Stem Cell Transplantation

While various peripheral blood stem cell (PBSC) mobilization strategies have been employed, granulocyte-colony stimulating factor (G-CSF) with or without plerixafor is widely utilized for its efficacy, safety and cost. To date, the recombinant G-CSF known as tbo-filgrastim has not been compared to traditionally used filgrastim for mobilization of PBSC or for engraftment after autologous stem cell transplantation (ASCT). Therefore, we sought to compare the efficacy and cost of tbo-filgrastim to filgrastim during mobilization and engraftment. Patients with lymphoma or plasma cell disorders undergoing G-CSF mobilization, with or without plerixafor, were included in this retrospective chart review. Patients receiving a second autologous transplant or mobilized with chemotherapy were excluded. Tbo-filgrastim and filgrastim were given at a dose of 10 mcg/kg daily for four days prior to PBSC collection on day 5. The adjunct use of plerixafor was determined by circulating CD34+ cells/μL on day 4 of mobilization. Post-transplant, each agent was given at a dose of 5 mcg/kg daily on day +7 and continued until neutrophil engraftment. The primary outcome was total collected CD34+ cells/kg. Secondary outcomes included total collection days, peripheral CD34+ cells/μL on day 4 of mobilization, and the adjunctive use of plerixafor. Post-transplant outcomes included time to neutrophil engraftment, the number of blood product transfusions required prior to engraftment, and events of febrile neutropenia. Cost minimization analysis was also conducted to determine direct cost-savings between the two G-CSF products. A total of 148 patients were included in the final analysis. Median patient age was 60 years, 81 (55%) were male and 86 (58%) received filgrastim. Patients in the filgrastim group had a higher median age compared to those receiving tbo-filgrastim (61.5 vs 56 years, p=0.01). There were no other statistically significant differences in patient demographics between the two groups. Patients receiving filgrastim collected a median of 5.565 x 106 CD34+ cells/kg compared to a median of 5.9 x 106 CD34+ cells/kg for the tbo-filgrastim group (p=0.47). There were no statistically significant differences in all secondary endpoints except a mean fewer days of collection in patients receiving tbo-filgrastim (1.4 vs 1.6 days, p=0.01). Patients receiving tbo-filgrastim had a mean cost savings of $761.46 per patient during mobilization and $353.46 per patient during engraftment. Tbo-filgrastim demonstrated similar CD34+ yield as compared to filgrastim in the setting of PBSC collection with no difference in secondary efficacy and safety outcomes. Furthermore, tbo-filgrastim utilization was associated with cost savings of approximately $1,115 per patient.Table 1Patient demongraphics and outcomes*, n = 148Tbo-filgrastimFilgrastimP valueDemographics Age (years)56.0 (29.0-72.0)61.5 (21.0-82.0)0.016 Gender, male33 (53.2%)48 (55.8%)0.867 Diagnosis0.472Plasma cell disorders∗41 (66.1%)62 (72.1%)Lymphoma Mobilization21 (33.9%)24 (27.9%)Mobilization G-CSF(n)6286 CD34 Day 4 (cells/μL)10.5 (0.0-88.0)12.5 (1.0-65.0)0.534 Plerxiafor use45 (72.6%)60 (69.8%)0.854 CD34 Day 5 (cells/μL)5.00 (1.0-292.0)43 (4.0-174.0)0.087 CD34 collected days* (x 106 cells/kg)4.61 (0.12-19.83)3.78 (0.68-16.34)0.150 Total collection days (x 106 cells/kg)1.41.60.010 CD34 infused (x 106 cells/kg)5.90 (0.12-19.83)5.56 (1.69-16.34)0.470Engraftment G-CSF (n)7078 CD34* infused (x 106 cell/kg)3.62 (2.24-10.84)361 (1.70-9.78)0.447 Transfusion*Packed red blood cells1.71.70.866Plalelets1.71.40.113 Ferbrile neutropenia48 (68.6%)53 (67.9%)0.935 Days to WBC engraftment‡11.511.70.251 Days to platelet engraftment‡17.718.50.244Note: G-CSF - granulocyte colony stimulating factor; WBC - white blood cells*All values are median unless otherwise specified.‡Plasma cell disorder includes multiple myeloma; Waldenstrom's macroglobulinemia; and amyloidosis.‡Value reported as mean. Open table in a new tab

Development and Usage of Algorithms Demonstrate Cost Savings and Increased Efficiency in Peripheral Blood Stem Cell Mobilization and Collection

Development and Usage of Algorithms Demonstrate Cost Savings and Increased Efficiency in Peripheral Blood Stem Cell Mobilization and Collection

造血幹細胞移植における各種Cell processing

造血幹細胞移植におけるcell processing 部門は，移植前後の最重要領域であり，血液型不適合移植時の血球除去などの細胞濃縮，血縁ならびに非血縁者間末梢血幹細胞採取(Peripheral blood stem cell collection; PBSCC)，移植後再発時などに行われるドナーリンパ球輸注(Donor Lymphocyte Infusion; DLI)，ドナー顆粒球輸注(Granulocyte Transfusion; GTx) などがあげられる．各々には目的となる細胞毎によるアフェレーシス機器と熟練の細胞採取技術者によって行われている．当院はこれらのすべてのcell processing を行う認定施設に選定されており細胞療法を発展させるべく積極的に実施している．

Peripheral Blood Stem Cell Collection with G-CSF Alone within 2 Weeks after Surgery in Patients with Pediatric Brain Tumors Requiring Craniospinal Irradiation

Peripheral Blood Stem Cell Collection with G-CSF Alone within 2 Weeks after Surgery in Patients with Pediatric Brain Tumors Requiring Craniospinal Irradiation

Collection of peripheral hematopoietic stem/progenitor cells.

Several variables possibly affecting collection of peripheral hematopoietic stem/progenitor cells (PBSC) were evaluated: type of apheresis machine (Amicus version 2.5, Baxter vs Cobe Spectra version 7.0, Terumo BCT), venous access (peripheral vein vs central venous catheter, i.g. CVC), and apheresis regimen (standard vs large volume leukapheresis, i.g. SVL vs LVL) with the objective to increase collection efficacy at the site. Peripheral blood represents the currently preferred source of hematopoietic stem/progenitor cells (HSCs) for transplantation. Data regarding 169 collection procedures performed in healthy donors and patients between January 2008 and December 2011 at the Clinics of Haematology and Transfusiology in St Cyril and Method Hospital in Bratislava (Slovakia) were analysed. With Cobe Spectra apheresis machine it was possible to process larger blood volumes per procedure with higher CD34+ cell collection efficiency (p = 0.0229) and lower RBC contamination of the harvest than with Amicus (p = 0.0116). On the other hand, Amicus helped to limit PLT contamination of the harvest (p < 0.0001), thus minimizing post-procedural decrease in patient´s PLT count. The highest detected advantage of CVC usage was higher flow rate of procedure, thus processing larger blood volumes per unit of time. Interesting finding was the tendency to lower harvest PLT contamination (p = 0.054). When LVL was performed, significantly higher HSCs yields were collected, even in "poor mobilizers" when the pre-run parameters were low. Management of PBSC collection requires a particular approach in each subject. Institutionally and individually optimized collection may help to improve the transplantation outcome and decrease the financial costs (Tab. 8, Ref. 15).

Delta-Neutrophil Index Is a Useful Indicator for Initiation of Peripheral Blood Stem Cell Collection in Chemo-Mobilized Patients

Introduction: The most powerful predictor of successful peripheral blood stem cell (PBSC) collection is the count of circulating CD34+ cell on the starting day of harvest. However, measurement of CD34+ cell requires flow-cytometry method and the decision of PBSC collection initiation usually does not depend on CD34+ count result in real practice. Delta-neutrophil index (DNI) is a calculated parameter from an automated blood cell analyzer (ADVIA 120, Siemens, Inc.) which shows the difference in leukocyte subfractions by cytochemical myeloperoxidase reaction and nuclear lobularity assay. We investigated on DNI as a potential indicator for initiation of autologous PBSC harvest.Materials and Methods: Total 114 patients who received chemo-mobilization in Severance Hospital from October 2010 to March 2014 were enrolled. All data were collected retrospectively from the patients’ medical records. Mobilization chemotherapy regimens were selected by the physician.G-CSF (600mcg/day) was administered from 1 to 3 days after end of chemotherapy until the last day of PBSC collection. Peripheral blood complete blood cell count and DNI was done daily from the end of chemotherapy. Measurement of CD34+cell was done only on the first day of harvest. Collection was initiated when the white blood cell (WBC) count rebounded > 3.0 x 109/L. We evaluated correlation of collected CD34+ cell count (/Kg) and DNI and tried to find the cutoff value of DNI for successful harvest.Results: PBSC collection data of total 114 patients were retrospectively reviewed (lymphoma n=96; multiple myeloma n=18). Mobilization regimen were variable (high dose etoposide n=71; high dose cyclophosphamide n=17; DHAP n=11; IVAM n=9; ICE n=2; high dose MTX+Ara-C n=3; other n=2). Median cycle number of previous chemotherapy line was 1 (range 1-5), and chemotherapy number 6 (1-18). 17 patients (14.9%) received radiation therapy. The median number of G-CSF was administered median 10 days (range 4-22). Peripheral blood CD34+ cell counting was done in 58 patients (50.9%). For 58 patients, WBC count, neutrophil count, mococyte count, DNI showed correlation with CD34+ cell (/mcL). Also interval between the day before harvest of WBC, neutrophil, monocyte showed correlation with CD34+ cell(/mcL). The highest correlation coefficient was shown with PB CD34+ count (ρ=0.876, P<0.001) followed by DNI (ρ=0.470, P<0.001), WBC interval (ρ=0.414, P<0.001), neutrophil interval (ρ=0.348, P<0.001), first day WBC (ρ=0.324, P<0.001). When successful first day apheresis was defined as > 1.0 x 106/kg, receiver operator characteristic (ROC) curve analysis showed DNI cutoff of 11.75 which had area under the curve (AUC) of 0.875 (sensitivity 81%, specifity 85.7%, P< 0.001). And if successful first day apheresis was defined as > 2.0 x 106/kg, receiver operator characteristic (ROC) curve analysis showed DNI cutoff of 13.75 which had area under the curve (AUC) of 0.845 (sensitivity 82.1%, specifity 76.7%, P< 0.001).Summary and Conclusion: DNI correlates with peripheral circulating CD34+ count on the first day of PBSC harvest after chemo-mobilization. And it has very high sensitivity and specifity for prediction of successful collection on first day. DNI may be used as useful marker for initiation and prediction of successful PBSC collection in chemo-mobilized patients. DisclosuresNo relevant conflicts of interest to declare.

Depletion of CD45RA+ Cells from Mobilized Peripheral Blood Stem Cell (PBSC) Collections As an Integral Part of Hematopoietic Stem Cell Transplantation (HSCT) and Cellular Therapy (CT)

Clinical scale depletion of CD45RA+ naïve T cells from leukapheresis collections can be achieved using CliniMACS (Bone Marrow Transplant. 2013, 1-7). In this study, PBSC collections depleted of CD45RA+ cells were infused as part of CT treatment protocols for pediatric patients with hematologic malignancies and solid tumors in 2 separate clinical trials. Between Sep 2012 and May 2014, a total of 25 CD45RA+ cell depletion procedures was performed for 25 patients. Mobilized PBSC leukapheresis units were collected (using Spectra) from HLA haplo-identical donors and held overnight in cold prior to cell selection. Characterization of the products before and after CD45RA+ cell depletion is shown in Table 1. Depletion of all CD45RA+ cell averaged to 3.39-log while depletion of CD3+CD45RA+ T cells was > 3.76-log, as this subset of cells became undetectable in 8 (out of 25) final products. CD34+ cell recovery was 65.4%, similar to that of CD3+CD45RO+ T cells (65.1%), which constituted > 99% of all T cells after depletion. From the PBSC collection, each patient received a median dose of 0.009 x 106 CD3+CD45RA+ cells/kg and 11.1 x 106 CD34+ cells/kg. The final product contained a substantial amount of CD14+ monocytes (median = 22.0%, range: 7.0 – 46.4), few CD56+ NK cells (3.1%, range: 1.3 – 8.1) and almost no CD19+ B cells (0.04%, range: 0.01 – 0.30). After processing the cells were resuspended in Plasmalyte + 5% human serum albumin and infused as soon as possible. There was no reported contamination (growth of bacteria – aerobic or anaerobic and fungus) in the final cell products and no infusion related adverse events were observed. In conclusion, we have demonstrated clinical scale depletion of naïve T cells can be achieved consistently and the final cell product can be safely infused into patients as part of HSCT and / or CT program.Table 1:Cell Populations (Median & ranges) Before and After CD45+ Cell DepletionBefore DepletionAfter DepletionTotal Nucleated Cells3.97 x 1010 (1.55 – 6.60)1.47 x 1010 (0.43 – 2.92)% CD34+ Cells0.85% (0.29 – 2.30)1.45% (0.32 – 4.63)% CD3+ T Cells22.8% (11.5 – 50.9)13.1% (3.9 – 37.7)% CD45RA+ Cells39.6% (27.2 – 64.1)0.05% (0.01 – 0.79)% CD3+ CD45RA+ Naïve T Cells12.3% (4.3 – 29.2)0.01% (0 – 3.42)% CD3+ CD45RO+ Memory T Cells6.5% (2.0 – 15.6)13.0% (3.9 – 37.5) DisclosuresNo relevant conflicts of interest to declare.

Remobilization with G-CSF Is Less Effective Than the Initial Mobilization in Healthy Donors Undergoing Peripheral Blood Stem Cell Collection for Allogeneic Transplantation

Background: The need to repeat peripheral blood stem cell (PBSC) mobilization and collection arises infrequently in healthy donors, but may be required due to insufficient initial collection, graft failure, or relapse of the recipient’s disease. Currently no published data exists on the efficacy of remobilization of healthy PBSC donors. Studies of remobilization in patients undergoing autologous transplantation (ASCT) have largely focused on the use of alternative mobilization agents such as chemotherapy or plerixafor. Boeve et al (Bone Marrow Transplant, 2004) reported that remobilization with G-CSF in patients undergoing ASCT who failed initial mobilization with G-CSF, resulted in higher numbers of CD34+ cells collected than the initial collection, though this required a doubling of the dose of G-CSF.Patients/Methods: We performed retrospective chart review of 977 consecutive adult (>18 yrs) donors who underwent apheresis for PBSC donation at Washington University School of Medicine from 1995 through 2013. We identified 66 donors who had undergone more than one mobilization. Two cohorts of donors were identified for analysis: Group 1 included donors mobilized initially and again subsequently with G-CSF (10 ug/kg/day), or GM-CSF (5 ug/kg/day) + G-CSF (10 ug/kg/day). Group 2 consisted of donors mobilized with a CXCR4 antagonist, plerixafor (240-320 ug/kg) or POL6326 (1000-2500 ug/kg), and subsequently were remobilized with G-CSF (10 ug/kg/day).Statistical Analysis: Spearman correlations were performed to analyze the relationship between peak peripheral blood (PB) CD34+/uL level; the number of CD34+ cells collected per kg (recipient weight); and the number of CD34+ cells per L of apheresis collected during initial mobilization (MOB1) and remobilization (MOB2); and the interval (days) between MOB1 and MOB2. One-way ANOVA with repeated measures analyses were performed to determine the relationship of PB CD34+/uL, CD34+/kg and CD34+/L during MOB1 and MOB2.Results: Group 1 included 30 donors. The median age was 49 years (range 18-75) and 15 were male. The median number of days between MOB1 and MOB2 was 140 (range 26-2238). All 30 donors were remobilized due to graft failure or relapse of the recipient’s disease. PB CD34+/uL, CD34+/kg and CD34+/L all correlated between MOB1 and MOB2. The mean PB CD34/uL at MOB1 was 69 compared to 37 at MOB2 (p= 0.029); the mean CD34/kg collected at MOB1 was 5.6x106 compared to 3.3x106 at MOB2 (p= 0.002); and the mean CD34/L collected at MOB1 was 24.0x106 compared to 17.6x106at MOB2 (p= 0.023). The interval between MOB1 and MOB2 did not correlate with any of the MOB2 variables. Results from the analysis are summarized in Table 1.Group 2 included 32 donors. The median age was 51 years (range 21-67) and 18 were male. The median number of days between MOB1 and MOB2 was 20 (range 4-1123). 18 donors were remobilized due to mobilization failure, while 14 were remobilized due to graft failure or relapse of the recipient’s disease. The mean PB CD34/uL at MOB1 was 15 compared to 68 at MOB2 (p< 0.001); the mean CD34/kg collected at MOB1 was 2.5x106 compared to 7.1x106 at MOB2 (p< 0.001); and the mean CD34/L collected at MOB1 was 10.6x106 compared to 30.1x106at MOB2 (p< 0.001). The interval between MOB1 and MOB2 did not correlate with any of the MOB2 variables. Results from the analysis are summarized in Table 2.Conclusion: Remobilization with G-CSF or GM-CSF and G-CSF after initial successful mobilization with the same regimen results in poorer mobilization while remobilization with G-CSF after initial mobilization with a CXCR4 antagonist results in dramatically improved mobilization. The reason for this remains unclear, but in this study the interval between collections was not associated with successful remobilization.Abstract 850. Table 1Group 1MOB 1MOB 2One-way ANOVASpearman CorrelationPB CD34/ul69 (13-417)37 (1-115)F(1.0, 29.0) = 5.26, p= 0.029r= 0.615, p< 0.001CD34/kg (x106)5.6 (0.8-13.8)3.3 (0.3-10.6)F(1.0, 29.0) = 11.77, p= 0.002r= 0.483, p= 0.007CD34/L (x106)24.0 (4.5-72.0)17.6 (2.8-41.3)F(1.0, 29.0) = 5.74, p= 0.023r= 0.566, p< 0.001Abstract 850. Table 2Group 2MOB 1MOB 2One-way ANOVASpearman CorrelationPB CD34/ul15 (2-54)68 (14-358)F(1.0, 31.0) = 23.16, p< 0.001r= 0.433, p= 0.013CD34/kg (x106)2.5 (0.2-19.7)7.1 (1.7-42.4)F(1.0, 31.0) = 33.84, p< 0.001r= 0.769, p< 0.001CD34/L (x106)10.6 (1.4-67.1)30.1 (6.0-165.0)F(1.0, 31.0) = 34.70, p< 0.001r= 0.774, p< 0.001 DisclosuresNo relevant conflicts of interest to declare.

Donor-to-Recipient Weight Ratio Is Independently Associated with CD34+ Yield in Healthy Donors Undergoing Peripheral Blood Stem Cell Collection for Allogeneic Transplantation

Background: Over the past two decades, peripheral blood stem cells (PBSC) have surpassed bone marrow as the preferred graft source for adult allogeneic transplantation due to its more rapid engraftment and potentially better graft-vs-tumor effects, and because PBSC collection is much less invasive for the donor. The optimal CD34+ PBSC dose is ≥ 4.0x106cells/kg, but doses ≥ 8.0x106cells/kg are suggested by some for reduced-intensity conditioning and haploidentical transplants. There is no established minimum CD34+ PBSC dose, but doses below 2.0x106cells/kg have been associated with a higher risk of engraftment delay and failure. There is significant inter-donor variability in the ability to mobilize PBSCs. Several factors have been identified as predictors of PBSC mobilization in healthy donors including: gender, age, weight, body mass index (BMI), and blood counts before and after mobilization. The impact of the donor’s comorbidities on mobilization is currently unknown.Patients/Methods: We performed retrospective chart review of 488 consecutive adult patients who underwent apheresis for allogeneic stem cell donation at Washington University School of Medicine from 2006 through 2013. Patients who received any collection regimen other than 10mcg/kg of G-CSF daily with 20 liters (+/-10%) apheresis on Day 5 were excluded. Patients who had undergone a previous apheresis for stem cell donation were excluded. Univariate analysis was performed to identify predictors of CD34+ PBSC collection in a single apheresis. Variables analyzed were: gender; age; weight; BMI; donor-to-recipient weight ratio; pre and post-mobilization blood counts (white blood count [WBC], hematocrit, platelets, neutrophils, lymphocytes, and monocytes); pre-mobilization glucose and triglyceride levels; post-mobilization peripheral blood (PB) CD34+count; and medical history significant for hypertension, hyperlipidemia, or diabetes mellitus. Subsequently, a linear regression multivariate analysis was performed with all variables found to be significant in the univariate analysis. 2-tailed tests for significance were used throughout the analysis.Results: 304 patients met the eligibility criteria for analysis. The median age was 53 years (range 18-76), 90% were Caucasian, and 50% were male. The median number of CD34+ cells collected was 7.4 x106/kg (range 0.8-27.1). 97% (295) collected ≥ 2.0x106 CD34+cells/kg, 81% (247) collected ≥ 4.0x106 CD34+cells/kg, and 44% (134) collected ≥ 8.0x106 CD34+ cells/kg. Post-mobilization PB CD34+ count (r= 0.841, p <0.001) and donor-to-recipient weight ratio (r= 0.439, p <0.001) were the strongest correlates with CD34+ collection. Weak correlations were seen with post-mobilization neutrophils (r= 0.360, p <0.001), WBC (r= 0.353, p <0.001), and hematocrit (r= 0.207, p <0.001); weight (r=0.280, p <0.001); BMI (r= 0.229, p <0.001); and age (r= -0.207, p <0.001). Male donors collected 9.1x106 CD34+ cells/kg compared to 7.5 x106 CD34+ cells/kg for females, on average (p = 0.003). Hypertension, hyperlipidemia, and diabetes mellitus were not associated with Day 1 CD34+ collection. We performed a multivariate model with the variables: post-mobilization PB CD34+ count; donor-to-recipient weight ratio; gender; post mobilization neutrophil, WBC, and hematocrit; weight; BMI; and age. In this analysis, post-mobilization PB CD34+count, donor-to-recipient weight ratio, and age were all independently significant.Conclusion: After one apheresis, the majority (81%) of donors collected ≥ 4.0x106 CD34+ cells/kg, the optimal number of CD34+ cells for standard allogeneic transplant, but only 44% were able to collect ≥ 8.0x106, the suggested number of CD34+ cells for reduced-intensity or haploidentical transplants. As these reduced-intensity and haploidentical transplants increase in frequency, the need for more accurate predictors of CD34+cell collection will also increase. Donor-to-weight ratio could be a useful tool to stratify potential donors for reduced-intensity and haploidentical transplants. In this study, 63% of donors with a donor-to-recipient weight ratio > 1.0 collected ≥ 8.0x106 CD34+cells/kg, while only 20% of donors with a donor-to-recipient weight ratio ≤ 1.0 did. While gender, weight, and BMI have previously been reported as predictors for CD34+ collection, they potentially were just surrogates for donor-to-recipient weight ratio. DisclosuresNo relevant conflicts of interest to declare.

Prospective Qualitative Study Concerning the Efficiency of PBSC Collections after a Change of Apheresis Device

BackgroundEfficiency measurement is an important indicator to improve quality of PBSC harvests. We have recently changed our apheresis device for PBSC collections: from the Cobe Spectra (Terumo BCT) to the Optia Spectra (Terumo BCT). We have further to this change decided to follow prospectively the harvests to estimate the efficiency of the new apheresis device.Material and MethodBetween November 2013 and April 2014, 21 consecutive harvests were conducted on Optia Spectra for ten consecutive patients/healthy donors. Six patients (3 multiple myeloma and 3 NHL) gave autologous PBSC. They were all mobilized with filgrastim (Neupogen, Amgen) which was associated with plerixafor (Mozobil, Genzyme) for two patients and pegfilgrastim (Neulasta, Amgen) for one patient. Four healthy donors gave PBSC for allogeneic HSCT; three were mobilized with filgrastim (Neupogen, Amgen) and one was mobilized with lenograstim (Granocyte, Chugai).ResultsThe data of our 21 consecutive harvests showed a median PB CD34+ count of 20/mL [range, 7-132] and a median collection of 1.94 CD34 +/kg [range, 0.48-8.33]. The correlation between the PB CD34+ count and the number of CD34+ collected was strong (R2=0.83) (fig.1). A median of 3 and 2 collections were necessary in order to obtain a suitable graft for autologous and allogeneic HSCT respectively.In this study, the efficiency of leukapheresis was only calculated for harvests with a PB CD34+ count > or = 20/µL (n=11), yielding a median efficiency of 30% (range, 13-52). These results were compared with those of our historic cohort of harvests achieved with the Cobe Spectra apheresis machine which yielded a median efficiency of 50% [range, 20-70]. In May 2014, in view of our lower efficiency results with the new device after 6 months of prospective study, we asked the Terumo BCT Company for an analysis of the harvests’ data recorded in the Optia Spectra apheresis machine.This analysis revealed a systematic accumulation of the buffy coat layer during the procedures, which led to the low efficiency of our harvests. Awareness of the fast accumulation of the buffy coat layer in the collection chamber, especially when the donor’s white blood cell count is > 30 G/L, is of utmost importance. The correction has to be made quickly by reducing the depth of collection from 60 to 40 (or 40 to 30) and by decreasing the speed of the pump to 3mL/min if the initial speed is > 3 mL/min. Theoretically, these two operations should allow the elimination of the buffy coat layer and improve the efficiency of the harvests. The outcomes of our patients are described in Table 1.ConclusionAfter 6 months of prospective study, the median efficiency of our PBSC harvests with the new device was 30%, that is below our expected value. Repeated procedures were necessary to collect enough CD34+ cells for grafting. For this reason, we requested that Terumo BCT make a retrospective analysis of the collection data recorded in the Optia Spectra system. Their audit showed a systematic accumulation of the buffy coat layer in the collection chamber. Now corrective measures have been implemented and we are going to continue this study over the next 6 months (June 2014 - November 2014) to estimate our new PBSC collections’ efficiency. [Display omitted] Abstract 5834 Table 1Patients outcome analysisnNber of collec- tion(s)CD34+ collectedDiseaseAuto/alloHSCTAplastic outgoing delayed (neutro or plat)Disease Status at+3MonthsDisease Status at +6Monthsalive at Last follow-up<!>Cause of death143.46MMautoNoPRPRYes<!226.82NHLautoNoCRNEYes338.21NHLautoNoCRCRYes425.65MMautoNoNENEYes5(1)8.33MDSalloNoCRCRYes6(2)4.31ALLalloNorelapseNENoRelapse7(2)2.1AMLalloYes (Plat)relapseCRYes8(2)4.6MF + AMLalloYes (Plat)NENENoRelapse941.03MMAutoNot realizedNENENEYes1014.09MMautoNoPRPRYes DisclosuresNo relevant conflicts of interest to declare.

Hypercvad Induction Chemotherapy Is Associated with Higher Rates of Stem Cell Mobilization Failure in Mantle Cell Lymphoma ( MCL)

Background: MCL patients deemed fit for high dose regimens undergo induction therapy with rituximab and chemotherapy (HyperCVAD/CHOP/bendamustine) followed by consolidation with ASCT (autologous stem cell transplantation). R-HyperCVAD is a dose intense regimen with high response rates for MCL when used in the upfront setting (ORR 97%; CR 38%; Romaquera JE, JCO 2005). R-bendamustine and R-CHOP are contemporary regimens with similar efficacy (ORR 85-93%; Rummel MJ, Lancet 2013). In a recent randomized phase II trial (SWOG 1106) accrual to the HyperCVAD arm was stopped due to higher than expected rates of peripheral blood stem cell (PBSC) mobilization failure.Methods: To assess the impact of these regimens on PBSC collection, we performed a retrospective analysis of newly diagnosed MCL patients undergoing induction chemotherapy with R-HyperCVAD versus R-CHOP/R-bendamustine and referred to transfusion medicine for PBSC collection prior to ASCT from 01/2009 to 12/2013. Patients were not allowed any salvage chemotherapy. The primary end point was successful stem cell collection defined as ability to collect ≥2.1 million CD34 cells/Kg. Secondary endpoints were number of days of apheresis, use of pleraxifor as mobilization salvage and total number of CD34+ cells collected.Results: A total of 91 MCL patients were eligible for analysis (Table 1). Patients who received HyperCVAD were younger at the time of collection (median: 56 vs 62 years; p <0.01) and were referred for collection earlier (median time from diagnosis to collection start: 4.5 vs. 6.4 months; p<0.01). There were no other baseline differences between the two groups in terms of gender, bone marrow involvement, stage of disease at presentation and use of plerixafor. While the median number of apheresis days were comparable: 4 (range: 1-9) for the HyperCVAD group and 3 (range: 2-8) for the other group (p=0.21), 18% patients in the Hyper-CVAD group failed to collect adequate numbers of PBSC (defined as < 2.1 ×10 6/Kg) compared to 4% in the other group (p=0.05). Additionally, the median number of CD34+ cells collected was lower in HyperCVAD group (p=0.05). After adjusting for baseline differences in age and timing of collection, for a patient who received HyperCVAD, the odds of failing to collect were 7.28 times higher (95% CI: 1.01, 52.57) than the odds for a patient who received a non-HyperCVAD induction regimen (p=0.05).Ultimately, 81/91 (89%) patients proceeded to high dose chemotherapy and ASCT [82% in HyperCVAD versus 96% in non-HyperCVAD (p=0.06)]. Four of the remaining 10 patients with mobilization failure (all from HyperCVAD arm) proceeded to Allogeneic HCT, the remaining 6 patients did not receive any further treatment.Conclusion: Patients with MCL receiving R-HyperCVAD chemotherapy in the frontline setting have a significantly higher rate of PBSC mobilization failure and collect significantly fewer CD34+ PBSCs when compared to patients treated with comparable regimens. R-HyperCVAD should be used with caution in patients with newly diagnosed MCL who are eligible for ASCT. Some patients failing mobilization may be salvaged with use of plerixafor.Table 1:Patient, Transplant CharacteristicsVariableHyper-CVAD, n=45Median (Range)N (%)Other, n=46Median (Range)N (%)Patient Gender Female Male9 (20) 36 (80)12 (26) 34 (74)Age at Collection Completion (years)56 (40 – 68)62 (36 – 74)Time from Diagnosis to Start of Collection (months)4.5 (2.3 – 65.3)6.4 (3.9 – 69.4)Stage at Diagnosis I II III IV1 (2) 3 (7) 2 (4) 39 (87)2 (4) 1 (2) 6 (13) 37 (81)Bone Marrow Involvement at Diagnosis No Yes Not Done9 (20) 35 (78) 1 (2)13 (28) 32 (70) 1 (2)Treatment after Chemotherapy Auto Transplant Allo Transplant No Transplant37 (82) 4 (9) 4 (9)44 (96) 0 (0) 2 (4)Mozobil Era (08/16/2009) Pre Post8 (18) 37 (82)3 (7) 43 (93)Mozobil Usage No Yes29 (64) 16 (36)30 (65) 16 (35)Number of Collections4 (1 – 9)3 (2 – 8)Total CD344.5 (0.3 – 100.5)5.3 (0.7 – 76.6)Total CD34 (Failure Rate) < 2.1 >/= 2.18 (18) 37 (82)2 (4) 44 (96) DisclosuresChen:Seattle Genetics: Honoraria, Research Funding, Speakers Bureau.

Hemostatic Disorders and Thrombotic Events in Multiple Myeloma Patients during Peripheral Blood Stem Cell Mobilization

Background: Patients (pts) with multiple myeloma (MM) have an increased risk of thrombosis. Previous studies have shown that 3.7–4.6% of pts undergoing peripheral blood stem cell (PBSC) mobilization developed thrombotic complications.Aims: The aim of the study was to investigate the blood coagulation status in MM pts during PBSC mobilization.Methods: 20 pts: 12 males, 8 females at the age of 27 – 64 years (median 53) were included in the study. After bortezomib-containig induction therapy 5 of them achieved CR, 11 – VGPR and 6 – PR. PBSC mobilization was performed by using cyclophosphamide (CY, 4g/m2) and granulocyte colony-stimulating factor (G-CSF, 5mcg/kg/day). A central venous catheter (CVC) was placed on the day before CY administration and heparin-sulphate continuous infusion (500 IU/hour) as thromboprophylaxis was started. Subcutaneous injections of G-CSF were administrated when WBS decreased less 1 x 109/l. The procedures of stem cell collection were carried out when the number PB CD 34+ cells exceeded 10 x 103/ml. Hemostasis analysis was performed 5 times, namely before (point 0) and 24 hours after heparin administration (point 1), the next day after CY infusion (point 2), before G-CSF administration (point 3) and on the day of PBSC collection (point 4). Condition of hemostasis was assessed by the results of activated partial thromboplasin time (APTT, normal rate: 25 – 38 sec) and thrombin generation test, namely by using endogenous thrombin potential (ETP, normal rate: 760 – 1450 nM*min) in every point.Hypercoagulation was considered in cases when APTT < 25 sec and ETP > 1450 nM*min. And hypocoagulation was estimated by data APTT > 38 sec and ETP < 760 nM*min. Time series analysis performed using TIMESERIES procedure in SAS 9.3 (SAS Institute Inc. Cary, NC). Significance level of alpha was set at 0.05.Results: Before heparin administration (point 0) APTT evaluation showed normal coagulation in 18 pts (90%) and hypocoagulation in 2 pts (mean 34 sec, 95% CI 32 – 36 sec) (Figure 1). ETP was normal in 14 pts and increased in 6 cases (mean 1441 nM*min, 95% CI 1304 – 1578 nM*min) (Figure 2). Pts in CR demonstrated no significant difference in coagulation status compared to pts in VGPR or PR. On the next day after CVC incorporation and starting thromboprophylaxis (point 1), APTT and ETP have significantly changed (p < 0,05) to hypocoagulability in response to heparin infusion: mean data APTT became 38 sec (95% CI 36 – 41 sec) and mean ETP - 938 nM*min (95% CI 801 – 1074 nM*min). On the next day after CY administration (point 2) APTT was statistically significantly decreased (p < 0.05) reaching 35 sec (95% CI 34 – 36 sec). ETP had also changed but to a lesser degree, the mean was 1079 nM*min (95% CI 929 – 1229 nM*min). Hemostasis at point 3 was characterized by APTT mean 38 sec (95% CI 37 – 40 sec) and ETP mean 1057 nM*min (95% CI 911 – 1203 nM * min). On the day of PBSC collection mean values of measured parameters have not changed as compared to the previous point: APTT was 40 sec (95% CI 37 – 43 sec) and ETP was 1046 nM*min (95% CI 916 – 1176 nM*min) (Table 1).Thrombotic events were diagnosed in 2 cases of the total of 20 pts. There were two females, 54 and 56 years, both in CR MM. The first pt developed CVC-associated thrombosis. She had additional risk factors such as obesity (BMI – 44) and heterozygous MTHFR polymorphism. The second pt demonstrated acute violation of cerebral circulation which arose due to occlusion of the right internal carotid artery. She was a hard smoker and heterozygous FV Leiden polymorphism. Having confirmed thrombotic complications, the dose of heparin had been increased. Besides, folic acid was administrated for the first pt and combination of aspirin and pentoxifilline for the second. PBSC collection was successful for both pts.Summary: The results of our study identify latent hypercoagulation status in 30% pts of confirmed by elevated ETP. Those changes were registered despite of achievement CR or PR MM. Thrombotic complications have developed in 10% from our group. Notably, one should consider individual risk factors for each pt. [Display omitted] [Display omitted] Table 1Measurements data of parameters of hemostasisMeasuring pointMean APTT (95% CI)Mean ETP (95% CI)033.6 (31.2-35.9)*1441.1 (1304.3-1577.9)*138.6 (35.7-41.6)*937.6 (801.3-1074.0)*235.2 (33.9-36.5)*1079.0 (928.8-1229.2)*338.1 (36.7-39.5)1057.2 (911.4-1203.1)439.6 (36.5-42.8)1046.2 (916.0-1176.3)* Means in the same column are significantly different (P<0.05) DisclosuresNo relevant conflicts of interest to declare.

LACE - an Effective Conditioning Regimen for Lymphoma Patients Undergoing Autologous Transplant- Analysis of Outcomes and Prognostic Factors

Background: High dose chemotherapy followed by autologous stem cell transplantation (ASCT) is the treatment of choice for patients with relapsed and refractory lymphomas. BEAM (BCNU, etoposide, cytarabine, melphalan) and CBV (cyclophosphamide, BCNU , VP-16) are most widely used conditioning regimens prior to ASCT in lymphomas. LACE has been found to be an effective regimen though outcome data is sparse.As BCNU was difficult to procure in India, we began using this regimen since November 2007. This study is a retrospective analysis to evaluate outcomes and possible prognostic factors in this cohort of patients.Methods: All patients between November 2007- January 2014 who received LACE regimen for primary progressive, chemotherapy sensitive relapse or relapsed-refractory Hodgkin’s (HL) and non- Hodgkin’s lymphoma (NHL) were included.Patients received salvage chemotherapy with either GDP, ICE, MINE or DHAP with or without rituximab (NHL patients). PET-CT was performed in all patients after 2-3 cycles of salvage chemotherapy. Response assessment was performed according to Cheson’s criteria. Patients underwent peripheral blood stem cell collection after 3rd or 4th cycle of salvage chemotherapy.Conditioning regimen used was LACE (lomustine-200 mg/m2 d-7, etoposide 1000mg/m2 d-7, ara-c 2000 mg/m2 d-6, d-5 and cyclophosphamide 1800 mg/m2 d-4 to d-2). PET-CT was done on day 100, 1 year post transplant and then yearly for next 4 years. Incidence and grade of treatment related toxicity was recorded according to CTCAE V-3. Prognostic factors evaluated for overall survival (OS) and progression-free survival (PFS) included time between diagnosis – transplant; time between CR1 - first relapse; baseline, presalvage and pretransplant serum albumin,LDH and B2 microglobulin; PET positivity pretransplant, at day 100 and day 360; remission status at time of transplant; IPI (NHL) and IPS (HL) at baseline and at relapse; stage at diagnosis and at relapse. Probabilities of OS and PFS were estimated using the Kaplan–Meier method. Univariate comparisons of survival times for potential prognostic factors were made using the log-rank test. Multivariate analysis of significant factors was performed by Cox –regression analysis.Results: Seventy patients had HL while 30 NHL (Male-73, Female-27) with median age at transplant of 23 years. In NHL cohort, 19 were DLBCL, 6 were T-cell lymphomas, 4 mantle cell lymphoma and 1 Burkitt’s lymphoma.The median time from complete remission (CR) to first relapse and time from diagnosis to transplant were 1.4 and 1.9 years respectively. The median number of lines of chemotherapy pre transplant was 2. GDP (53 patients) was the most commonly used salvage chemotherapy. At the time of transplant, 68% were in CR, 29% in partial remission (PR) and 3% had refractory state. The incidence of grade 3-4 oral mucositis was 8% with a median duration of 3.5 days. The incidence of grade 3-4 diarrhea was 4% with median duration of 3 days. Median days to myeloid and platelet engraftment were 10 and 13 respectively. The median follow-up was 2.9 years. The probability of OS and PFS at 5 years was 67% and 57% for the whole group, 73% and 62% in HL group and 51% and 46% in NHL group. Seven patients died due to transplant related causes which included 6 due to sepsis and 1 due to herpes simplex encephalitis. In univariate analysis for OS, PET negativity pre transplant (P= 0.03), at day 100 (P= 0.019) and at day 360 (P= 0.01) were associated with better OS. Similarly, HL patients with IPS 0-2 (P=0.002) at time of relapse had better OS. Univariate analysis for PFS showed that PET negativity pre transplant (P=0.002), at day 100 (P= 0.0001) and at day 360 (P=0.00) was associated with better PFS. HL patients with IPS 0-2 (P=0.000) and NHL patients with IPI 0-2 at time of relapse (P=0.007) had better PFS. Patients in CR at the time of transplant had better PFS than those in not in CR (P=0.008). Overall HL patients had better OS (P=0.037) and PFS (P=0.097) compared to NHL group. Multivariate analysis for OS (P= 0.021) and PFS (P=0.001) revealed PET negativity at day 100 as the only significant prognostic factor.Conclusions: This is the largest reported cohort of lymphoma patients transplanted with LACE regimen. LACE is effective and well tolerated conditioning regimen in lymphoma transplant. PET negativity at various time points pre and post transplant prognosticates for better survival. DisclosuresNo relevant conflicts of interest to declare.

Defibrotide for Prophylaxis of Hepatic Veno-Occlusive Disease in Autologous, Non-Purged Peripheral Stem Cell Transplantation for High Risk Neuroblastoma Patients

High-dose chemotherapy (HDC) represents the standard of treatment for high-risk neuroblastoma(NBL), hepatic veno-occlusive disease (VOD) is a common, 10-50% and serious complication of haematological stem cell transplantation (HSCT), with up to 90% mortality rates. We planned study to assess whether the use of prophylactic defibrotide in paediatric patients who were heavily treated with chemo-radiotherapy before transplant, and then underwent autologous HSCT.Seventeen patients who underwent autologous, unpurged peripheral stem cell transplantation PBSC with a high risk of developing VOD, between January 2003-July 2014, were given Defibrotide prophylaxis 25mg/kg/day for 30 days, commencing on -1 of conditioning regimen. All patients were stratified by INSS stage, age, N-MYC status. All were treated with six cycles of induction chemotherapy, myeloablative intensification, surgery for primary site, radiation therapy to the primary tumour site plus metastatic sites i.e bone metastases including skull. Meta-iodobenzylguanidin treatment as targeted radiotherapy was given on -21 prior to myeloablative chemotherapy. Oral 13-cis retinoic acid was employed on day +90 post-transplant. CD34+ cell mobilization and PBSC collection was carreid out after two-four cycles of induction cycles. Conditioning regimen and stem cell infusion was done following four-six weeks of last chemotherapy cycle, in order to reduce the toxicitiy. Conditioning regimen CEM consisted Carboplatin, Etoposide, Melfalan. There were no toxic deaths. All of the patients receieved antimicrobial prophylaxis and total parenteral nutrition support when was needed. Myeloid engraftment on day+13, erythroid engraftment on day+18 and thrombocyte engraftment was achieved on day +23.Defibrotide prophylaxis seems to reduce incidence of VOD and is well tolerated. VOD incidence and severity was reduced in the defibrotide group which suggests that defibrotide might be effective in preventing and treating VOD. Sufficiently powered randomised trials are now required to definitively test the role of defibrotide in this setting. DisclosuresNo relevant conflicts of interest to declare.

Brentuximab Vedotin (BV) Does Not Impair Autologous Peripheral Blood Stem Cell Mobilization and Collection in Refractory/Relapsed CD30+ Lymphoma Patients Undergoing High-Dose Chemotherapy and Autologous Stem Cell Transplant (HDC-ASCT)

Introduction. Salvage chemotherapy followed by HDC-ASCT is considered standard of care for chemosensitive patients who have relapsed after initial therapy. CD30 is commonly expressed in Hodgkin Lymphoma (HL) and in some cases of non-Hodgkin lymphoma (NHL). Brentuximab vedotin (BV), an antibody-drug conjugate that targets CD30, induces high response rates and is now used in the salvage setting prior to HDC-ASCT. It is not clear if BV use peri-mobilization would influence mobilization and collection of autologous CD34+ stem cells. We therefore examined 42 patients who were treated with BV prior to HDC-ASCT.Methods. We retrospectively reviewed the HDC-ASCT databases of University Hospitals Case Medical Center (UHCMC) and MD Anderson Cancer Center (MDACC) and identified 42 patients who were treated with BV prior to HDC-ASCT between February 2009 and April 2014. The median age was 37 years (range, 18-67) and 52% (n=22) were male. Diagnoses were HL (n=30; 71%;), and NHL (n=12; 29%; anaplastic large cell, n=6; diffuse large B-cell, n=3; unknown subtype, n=3). Median times from diagnosis to transplant, from initial BV treatment to transplant and from last BV treatment to stem cell collection were: 21 months (range, 10-210), 5 months (range, 1.5-16.8), and 30 days (range, 2-280), respectively. Our subjects had failed multiple conventional treatments with a median of 3 (range, 2–8) lines of treatment before HDC-ASCT; 38% (n=16) received involved field radiation therapy. BV was given at 1.8 mg/kg IV every 21 days. Median number of BV cycles was 4 (range, 1-16) and the overall response rate to treatment was 71% (CR 55% + PR 16%). Thirty patients (71%) were in complete remission (CR) at the time of transplant (CR2 = 6; CR≥3 = 24), 4 (10%) were in partial remission (PR) (PR2 = 1; PR≥3 = 3), 6 patients (14%) had stable disease and 2 patients (5%) were transplanted with progressive disease. Stem cell collection target was 5 x 106 CD34+ cells/Kg. Mobilization regimens used were chemotherapy/G-CSF-based in 32 patients (76%) and Plerixafor/G-CSF-based in 10 patients (24%). Use of chemotherapy/G-CSF in first mobilization was standard at MDACC, whereas plerixafor/G-CSF was used as first mobilization at UHCMC.Results. Thirty-nine (92.8%) of 42 patients were successfully mobilized on the first attempt. Second mobilization was required in 3 cases (7.1%). Second mobilization regimens included Cyclophosphamide/G-CSF (n=2) and Plerixafor/G-CSF (n=1). The median number of infused CD34+ cells was 5.46 x106/kg (range, 1.65-54.78 x106/kg). All patients engrafted neutrophils and platelets at a median time of 10 days (range, 9-13), and 10.5 days (range, 7-35), respectively. The median time to RBC transfusion independence was 8 days (0-34). With a median follow-up of 12 months (range, 0–63), day 100 treatment-related mortality was 0%. The one-year actuarial event-free and overall survival is 50.5% and 84.1%, respectively.Conclusion. Within the limitations of this retrospective study, BV before HDC-ASCT did not adversely affect peripheral blood stem cell mobilization, collection and engraftment in a cohort of heavily pre-treated, relapsed/refractory patients with CD30+ lymphomas. DisclosuresCaimi:Seattle Genetics: Equity Ownership.

Lower Doses of Plerixafor Use Is Sufficient to Effectively Mobilize PBSC (Peripheral Blood Hematopoetic Stem Cells) When Compared to Standard Administration

Background: Plerixafor (CXCR4 chemokine receptor antagonist), is indicated to mobilize hematopoietic stem cells (HSCs) to the peripheral blood for collection and subsequent autologous transplantation in patients who are poor mobilizers. In this retrospective analysis, we compared the efficacy of plerixafor 0.24 mg/kg by subcutaneous (SC) injection (1-2 doses) versus plerixafor (3-4 doses) 0.24 mg/kg by SC injection in terms of their effect on the hematopoietic recovery in hematopoietic cell transplantation (HCT).Methods: We retrospectively reviewed the peripheral blood stem cell (PBSC) collections of 40 consecutive lymphoma and multiple myeloma patients who underwent autologous HCT, who had previously mobilized poorly and were given plerixafor for mobilization. Twenty-six patients received 1-2 doses of plerixafor and 14 patients received 3-4 doses. All patients gave written informed consent for the procedure. Successful engraftment time was defined as neutrophils >0.5x 109 cells/L, platelets>20 x109 cells/L for three consecutive days. SPSSincver. 20 is used for statistics.Results: All of the 40 patients were mobilized promptly after plerixafor administration without any problem. The mean amount of autologous CD34+ PBSC was 3,8 x 10(6)/kg versus 3,2 x 10(6)/kg, for 1-2 doses of plerixafor administration ve3-4 doses, respectively(p=0,4). Median times to the neutrophil engraftment to >0,5x 109/L were 12 and 10 days for the 1-2 doses and 3-4 doses, respectively (p=0,2). Additionally, median times to the platelet engraftment to >20x 109/L were 14 and 15 days for the 1-2 doses and 3-4 doses, respectively (p=0,6).Conclusions: These data show that the lower doses, i.e. 1-2 days of plerixafor administration evaluated in this study are sufficient to mobilize HSCs. This retrospective comparison suggest that larger prospective studies comparing two days vs. four days of plerixafor should be performed in order to further elucidate the role of two days of plerixafor use in HSC mobilization, and hence would a relatively cost effective way of mobilization in poor mobilizing patients. DisclosuresNo relevant conflicts of interest to declare.

What Can We Do for the Unrelated Donors of Hematopoietic Stem Cell to Lessen Their Agony?

Objectives: The study purposes are as follows. What are the incidences of adverse events (AEs) after hematopoietic stem cell donations among the unrelated donors registered in Korea Marrow Donor Program (KMDP)? What donor factors have influence on their general discomfort and the limitation of daily activity at the time of stem cell collection? What prohibits the donors from donating stem cell repetitively as a Good Samaritan?Methods: 1,666 consecutive unrelated donors were registered at KMDP from November 2007 to October 2013. General discomfort and pain were assessed using a visual analog scale (numerical measurement, 0 to 10) and other donor symptoms were assessed by making either yes or no reply to each item. The limitation of daily activity was assessed by numerical measurement (0 to 10 scales), 10 being the worst activity. The donor’s intention of repetitive donation was assessed by yes or no reply. The donor symptoms and the intention of repetitive donation were assessed within 48 hours after stem cell collection, and then at 1 week, 4 weeks, and 4 months thereafter.Results: There were 1,292 males and 374 females. 1,525 donors underwent peripheral blood stem cells (PBSC) collection, 100 bone marrow (BM) harvest and 40 leukapheresis for donor leukocyte infusion (DLI). All of 1,525 PBSC donors received granulocyte-colony stimulating factor (G-CSF). 76 donors underwent central line insertion for PB cells collection. The degree of general discomfort and the limitation of daily activity were analyzed in variables of sex, stem cell source, G-CSF administration, collection method, collection period and the intention of repetitive donation. A multiple linear regression analysis showed that general discomfort at the time of stem cell collection was significantly influenced by sex and collection period (P<0.05). The limitation of daily activity at the time of stem cell collection was significantly influenced by sex, stem cell source, and collection method (P<0.05). A multiple logistic regression analysis showed that the intention of repetitive donation at the time of stem cell collection and thereafter was significantly influenced by sex only. At least 80% of stem cell donors had the intention of repetitive donation.Compared to the donors using peripheral line, the donors using central line for PB cells collection suffered more frequently from various AEs: C-line site discomfort, hands and feet numbness, perioral numbness, dizziness, chills, nausea/vomiting and abdominal pain.Conclusions: Stem cell collection via peripheral line is recommended to alleviate the physical stress and to improve daily activity and to encourage repetitive donation, and female donors are more sensitive to stem cell collection than male donors. DisclosuresNo relevant conflicts of interest to declare.

A Single Centre Analysis of Spectra Optia Stem Cell Collection Data from Multiple Myeloma and Lymphoma Patients, with and without Plerixafor

Introduction:The Spectra Optia Apheresis System has been in use in our centre for 3 years for the collection of autologous peripheral blood stem cells (PBSC).We compared the Collection Efficiency (CE2) of the two Optias we have in use and also compared the CE2 of patients with Multiple Myeloma (MM) and Lymphomas, and those mobilised with and without Plerixafor. The Collection Efficiency (CE2) is calculated as follows: (Total CD34+ collected)/ (Whole Blood volume processed x Pre-CD34+ count). It is a crude estimation of the potential yield of a harvest and assumes that the CD34+ concentration is constant throughout a procedure. As the concentration of CD34+ cells in the peripheral blood will naturally drop off during a procedure, the CE2 can be expected to give a result that underestimates the yield. The pre-harvest FBC would normally be taken about 3 hours prior to commencement of the harvest, thus the peripheral CD34 may not fully reflect the count immediately before starting the harvest and this may also influence the accuracy of the CE2 result.Patients and Methods:Data on 143 peripheral blood autologous stem cell harvests from 87 patients collected using the Spectra Optia from mid-2011 to mid-2014 have been examined. Procedures without pre-harvest CD34+ counts or incomplete were excluded from the analysis. Procedures included in the analysis (138) are summarised in table 1. Median age of patients was 59 years (range 26-69). All patients were mobilised using disease specific standard protocols. Patients who fail to mobilise by the expected date were frequently given Plerixafor. A CD34+ dose of 5 x 106/kg was used as desirable target for reinfusion with enough for two reinfusion collected for MM patients to allow for repeat reinfusion if required.Statistical Analysis:Statistical analysis was performed using SPSS package. We have compared the CE2 achieved by each Optia machine, the CE2 results of the 2 main diseases MM and Lymphomas and to determine if any difference in Collection Efficiency is evident when patients are mobilised using Plerixafor.Results:143 collections were included in the analysis. Fifty patients underwent one collection, 27 patients underwent two collections, and 10 patients required three collections to achieve the minimum CD34+ cell dose required. The mean CE2 for the entire cohort was 74.23% and mean CD34+ yield was 4.6 x 106/kg/collection. There were no statistically significant differences in pre-collection CD34+ count, collection efficiencies and final CD34+ yield between groups according to instruments used, administration of Plerixafor and diseases (Table 1). A significant correlation between the Preleukapheresis peripheral blood CD34+ cell count and the CD34+ cell yield of each leukapheresis was shown by a linear regression analysis (r2 = 0.867, p < 0.0001; Figure 1).Conclusions:For maximisation of PBSC yield, the timing for PBSC collection after these mobilizations is critical. Among criteria utilized to predict yield, the circulating peripheral blood CD34+ cell count has been a good predictor of PBSC yield mobilized with protocols used in our centre. In our data, pre-leukapheresis peripheral blood CD34+ cell count on the day of collection correlates with the final CD34+ yield. Interestingly, administration of Plerixafor in patients clinically at risk of poor mobilisation allows collection of comparable quality to patients who mobilised without Plerixafor.Abstract 3848. Table 1Characterisation of collections, including device used, disease indication and use of Plerixafor.CD34_CE2Periph_cd34Actual yieldMeanSDNumber of proceduresMeanSDNumber of proceduresMeanSDNumber of proceduresOptia180.89%85.37%6251.843.9624.7334.37962268.69%17.38%7349.748.4734.6303.83473diseaselymphoma73.66%28.94%5751.157.2574.5814.60857myeloma75.39%76.69%7352.736.5735.0083.65173other65.45%11.13%515.412.05.945.6775Plerixaforno73.79%64.18%10755.147.61075.0784.208107yes76.19%35.77%2833.736.6283.1443.15528Total74%13550.6046.201354.6804.080135 [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Baseline Symptoms, Female Sex, and Younger Age Are Correlated with Higher Levels of Peri-Collection Pain, Symptoms, and Persistent Discomfort One Year after Related Donor BM and PBSC Donation: An Analysis of the Related Donor Safety Study (RDSafe)

Prospective data regarding donation related toxicities and time to recovery in related donors (RDs) of bone marrow (BM) and peripheral blood stem cells (PBSC) are limited, in contrast to significant data available regarding unrelated donor experiences. To address this lack of data in RDs, the NHLBI-funded Related Donor Safety Study (RDSafe; NCT00948636) prospectively enrolled RDs of all ages between 2010 and 2013 at 54 transplant centers in the United States. RDs were assessed for pre-donation comorbidities and health status, and then followed for 1 year after donation, collecting detailed information on adverse events, pain levels and 10 collection-related NCI-CTC symptoms. This report describes baseline, peri-donation, and through 1 year post pain/symptoms for RDs aged 18-60: 124 BM (38 centers, med age 33, 48% female) and 919 PBSC (42 centers, med age 49, 44% female).Results: About 20% of RDs reported pain and NCI-CTC symptoms at baseline, mostly grade 1, with females reporting pain/symptoms more often than males (see figures). Pain occurred in approximately 80% of donors surrounding collection with other symptoms occurring in 50-70% of donors. Pain and symptoms persisted in 10-20% of RDs at 1, 6, and 12 months; pain/symptoms returned to baseline in only a few categories assessed, but mostly remained elevated from baseline at 1 year. In addition, for both males and females, at 1 year grade 2-4 pain or symptoms were 2-3 times baseline rates. Multivariate analysis was performed on PBSC donors, where numbers were sufficient to look at key risk factors (see table). Females had more grade 3-4 pain at collection, and grade 2-4 pain and symptoms at 1 year. Age differences were noted, with RDs age 30-39 experiencing the highest amount of pain and symptoms and RDs age 50-60 experiencing less pain compared to donors aged 18-29. Pain or symptoms at baseline were important predictors of higher levels of reported pain/symptoms, respectively, during the collection and also higher risk of grade 2-4 pain at 1 year.Conclusions: One in 5 RDs have mild/moderate pain and/or symptoms at baseline, and the presence of pain/symptoms at baseline increases risk for experiencing higher levels of pain/symptoms during collection. Although the majority of RDs return to baseline status within a month of donation, >10-20% of RDs have lingering pain/symptoms 6-12 months after donation, and rates of grades 2-4 pain and symptoms at 1 year are more than double baseline. Females have higher grades of pain with collection and more grade 2-4 pain and symptoms at 1 year compared to males. RDs should be informed of the risk of mild/moderate pain/symptoms lingering through the first year after donation. Ongoing efforts to correlate baseline comorbidities of RDs with outcomes should allow RDs to be better informed of risk and potentially identify clinical risk profiles where RDs should be deferred.*Pain = max grade 2-4 or 3-4 2 days post-donation of BM or day+5 of PBSC collection (collection day 1).**Symptoms = fever, fatigue, rash, local site reactions, nausea, vomiting, anorexia, insomnia, dizziness, and syncopeTable 1MV Analysis of PBSC RDs for pain and donation-related symptoms: Odds Ratio (p-value).Pain*Symptoms**day+5 Grd 2-4day+5 Grd 3-41yr Grd 2-4day+5 Grd 2-41yr Grd 2-4Female1.667 (0.010)1.667 (0.016)1.887 (0.041)Age(0.001)(0.008)(<0.001)Age 18-291.01.01.0Age 30-390.76 (0.306)2.2 (0.036)2.48 (0.008)Age 40-490.68 (0.115)1.46 (0.283)1.49 (0.216)Age 50-600.45 (<0.001)0.93 (0.836)0.93 (0.830)Baseline pain overall(0.001)(0.004)(<0.001)Baseline pain Grd 01.01.01.0Baseline pain Grd 12.2 (<0.001)2.6 (0.06)2.95 (<0.001)Baseline pain Grd 2-44.73 (<0.001)4.4 (0.002)3.95 (<0.001)Baseline sx overall(<0.001)Baseline sx Grd 01.0Baseline sx Grd 1-42.16 (<0.001) [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.