Large Volume Leukapheresis Procedures Research Articles

Collection efficiency and safety of large-volume leukapheresis for the manufacturing of tisagenlecleucel.

In patients with relapsed or refractory B cell acute lymphoblastic leukemia or B cell non-Hodgkin lymphoma (r/r B-ALL/B-NHL) with low CD3+ cells in the peripheral blood (PB), sufficient CD3+ cell yield in a single day may not be obtained with normal-volume leukapheresis (NVL). Large-volume leukapheresis (LVL) refers to the processing of more than three times the total blood volume (TBV) in a single session for PB apheresis; however, the efficiency and safety of LVL for manufacturing of tisagenlecleucel (tisa-cel) remain unclear. This study aimed to investigate the tolerability of LVL. We retrospectively collected data on LVL (≥3-fold TBV) and NVL (<3-fold TBV) performed for patients with r/r B-ALL/B-NHL in our institution during November 2019 and September 2023. All procedures were performed using a continuous mononuclear cell collection (cMNC) protocol with the Spectra Optia. Although pre-apheresis CD3+ cells in the PB were significantly lower in LVL procedures (900 vs. 348/μL, p < .01), all patients could obtain sufficient CD3+ cell yield in a single day with a comparably successful rate of final products (including out-of-specification) between the two groups (97.2% vs. 100.0%, p = 1.00). The incidence and severity of citrate toxicity (no patients with grade ≥ 3) during procedures was not significantly different between the two groups (22.2% vs. 26.1%, p = .43) and no patient discontinued leukapheresis due to any complications. LVL procedures using Spectra Optia cMNC protocol was well tolerated and did not affect the manufacturing of tisa-cel.

Evaluating the efficiency and safety of large-volume leukapheresis using the Spectra Optia continuous mononuclear cell collection protocol for peripheral blood stem cell collection from healthy donors: A retrospective study.

Large-volume leukapheresis (LVL) refers to processing of more than three volumes of blood in a single session for peripheral blood stem cell collection. Recently, continuous mononuclear cell collection (cMNC) protocol has been developed using the Spectra Optia system, which is a widely used apheresis device. LVL using the novel protocol has been investigated in patients. However, the efficiency and safety of LVL in healthy donors using this protocol has not been characterized. Therefore, this study aimed to evaluate the efficiency and tolerability of CD34+ collection of LVL with the cMNC protocol in healthy donors. We retrospectively collected data on LVL (>3 total blood volume) and normal-volume leukapheresis (NVL) performed in healthy donors between October 2019 and December 2021. All procedures were performed using the cMNC protocol. Although pre-apheresis CD34+ cell count was lesser in LVL (23.5 vs. 58.0/μL, p < .001), CD34+ collection efficiency was comparable between LVL and NVL (61.2% vs. 61.4%, p = .966). Platelet loss was significantly higher in LVL compared to NVL (38.0% vs. 29.4%, p < .001), with no correlation between attrition of platelet and processing blood volume. Moreover, the incidence of citrate toxicity during procedures was comparable between the two groups (31.6% vs. 21.4%, p = .322). All LVL procedures could be completed without any adverse events. Allogeneic LVL procedure using Spectra Optia cMNC protocol was well tolerated by the donors and resulted in efficient collection of CD34+ cells, which was comparable to that of NVL.

"Tailoring" Hematopoietic Progenitor Cell Collection: Impact of a Data-Driven Prediction Algorithm for Blood Volume Processing in Large Volume Leukapheresis

Background:The predominant source of hematopoietic progenitor cells (HPCs) for hematopoietic stem cell transplantation (HSCT) is peripheral blood (PB). HPCs are collected for HSCT using large volume leukapheresis (LVL) that typically involves processing >4 blood volumes to ensure adequate yield of HPCs. LVL is time-intensive, causes volume overload and hypocalcemia, and in small children, requires prolonged sedation. The purpose of this quality improvement project was to assess whether a collection efficiency-based, data-driven prediction algorithm for HPC collection can meaningfully decease the volume of blood processed in well-mobilized patients.MethodsWe evaluated the CD34+ cellular collection efficiency 2 (CE2; see below) in 240 consecutive adult autologous (A-auto), 59 adult allogeneic (A-allo), and 23 pediatric autologous (P-auto) collections ("training set"). In the training set, mean (1SD) CD34+ CE2s in A-auto, A-allo and P-auto collections were 47% (16%), 51% (14%), and 59% (32%), respectively (Fig. 1).CD34 Collection Efficiency 2 (CE2) was calculated as below:CD34+ CE2= CD34+ yield/[BVP x Precollection PB CD34 count]Blood Volume to be processed (BVP) was calculated as below:BVP= CD34+ cell target/ [CD34+ CE2 x Precollection PB CD34+ count]In order to have a high level of confidence that the targeted dose of CD34+ cells will be collected with implementation of the prediction formula, the CD34+ CE2 was set at the 3rd percentile (30%) in the A-auto group, and lowest attained CD34+ CE2 in the A-allo (30%) and P-auto groups (25%) for the A-auto, A-allo and P-auto prediction formulae, respectively. These CD34+ CE2s were used to calculate the BVP in a large cohort of consecutive patients from Froedtert Memorial Lutheran Hospital and the Children's Hospital Wisconsin which included 461 A-Auto, 90 A-Allo and 16 P-Auto collections over a period of 16 months (2/2015- 5/2016). Data was collected from both prediction-driven truncated and standard non-truncated LVLs and comparisons were made with regards to blood volume processed, duration of LVL, financial impact and time to neutrophil and platelet engraftment.ResultsAfter implementation of the BVP prediction formula, 91 truncations were performed in 42 (9%), 41 (46%) and 8 (50%) of 461 A-auto, 90 A-allo and 16 P-auto collections, respectively, representing an overall truncation rate of 16% (Fig. 2). Of the 91 total procedures truncated, 4 procedures (all A-Auto) failed to reach the CD34+ collection target, however, cell yields were extremely close to targets and were not deemed to be prediction failures (Table 1). In truncated LVL procedures, reduction in volume of blood processed was 34% overall, with average decreased processing of 31%, 36% and 46% (of 4 blood volumes) in A-auto, A-allo and P-auto collections, respectively (Fig. 3). The average time for a randomly selected group of 20 standard 4BV collections was 4 hr 49 mins. A 34% truncation in BVP would result in the procedure concluding 1 hr 38 mins earlier and this prediction formula based intervention is predicted to save on average $132 per patient ($35/hr Nursing time; $25/hr Cell Processing technologist time, and 35% overhead) and total savings of $12,039 over 91 truncated procedures during the 16 month period assessed. Median neutrophil and platelet engraftment were not significantly different in autologous HSCT patients with truncated vs. non-truncated procedures (10 days each for neutrophil engraftment, and 20 days vs. 19 days in truncated vs. non-truncated procedures for platelet engraftment).ConclusionThe use of a data-driven HPC prediction algorithm resulted in the truncation of 16% of all LVL procedures. The volume of truncation was significant in most cases and did not adversely impact the ability to collect the desired number of stem cells and resulted in comparable neutrophil and platelet engraftment time. This approach may be particularly important in mitigating risks of LVL in populations with higher concern for adverse events such as children, and patients with poor tolerance for LVL-induced volume overload. Finally, in our experience, this approach has resulted in enhancing patient satisfaction associated with shorter procedures and promotes optimal use of healthcare resources by limiting costs. [Display omitted] DisclosuresPadmanabhan:Bloodcenter of Wisconsin: Patents & Royalties: A patent application has been filed on a Method of detecting platelet activating antibodies that cause heparin-induced thrombocytopenia/thrombosis; PCT/US14/62591; Terumo BCT: Consultancy, Honoraria; Schlesinger & Associates: Consultancy, Honoraria; LEK Consulting: Consultancy, Honoraria; Mallinckrodt Pharmaceuticals: Consultancy, Honoraria; Fenwal (Fresenius Kabi): Research Funding.

Autologous transplant in the treatment of severe aplastic anemia – A case report

The initial use of immunosuppressive therapy (IST) in severe aplastic anemia (sAA) or reapplication of IST-centered methods following disease relapse is successful only in well-selected patients. The potential treatment by autologous stem cell (SC) transplant in sAA is still an innovative/pioneering therapeutic approach. To our best knowledge, this is the second published case of autologous SC transplant in sAA. The aim of this work was to optimize mobilization and timing for SC harvesting – using our own controlled-rate cryopreservation – with higher CD34 +/CD90 + subset yield and recovery in order to obtain complete and long-term hematopoietic reconstitution following autologous SC transplant. We report a 35 year-old sAA male patient who initially underwent IST using rabbit ATG and Cyclosporine A (CsA). He was supportive transfusion dependent for the whole period of IST-phase. After the second IST-cycle, polymorphonuclear (PMN) cell count increase (>2.0 × 10 9/L) was observed, when SC mobilization, two large volume leukapheresis procedures and following autologous transplant were performed. The yields of harvested CD34 + and CD34 +/CD90 + cells were 5.75 × 10 6/kgbm and 1.7 × 10 6/kgbm, respectively. The quantity of applied CD34 + and CD34 +/CD90 + cells in autologous SC transplant were 5.45 × 10 6/kgbm (7-AAD CD34 + viability = 95.42%) and 1.63 × 10 6/kgbm (7-AAD CD34 + /CD90 + viability = 95.42%), respectively. Hematopoietic reconstitution registered due to second month after autologous SC transplant and he is 24 months in complete medullar, hematological and clinical remission, with normal cytogenetic status – applying only continuous CsA therapy. The results obtained strongly confirm that in sAA, with no allogeneic SC donor, autologous transplant can result in a successful clinical outcome. We suggest that CD34 +/CD90 + subset count in peripheral blood and/or cell-harvest could be more valuable predictive factor than total CD34 + quantity of optimized collection-timing and superior treatment efficacy of autologous SC transplant in sAA.

Peripheral Blood Progenitor Cell Collection in Low-Weight Infants (36 patients; 69 Large Volume Leukapheresis Procedures)

Peripheral blood progenitor cells (PBPC) have increasingly become the preferred source of stem cells for autologous transplantation due to their easier accessibility, rapid engraftment and higher safety. The harvest of PBPC in paediatric patients assumes critical importance, particularly in low-weight infants, in whom optimizing the procedure allows the collection of sufficient number of cells, with a reduced number of leukapheresis. The published data of these low-weight patients are scarce, and very likely the accumulation of cases will contribute to optimize the procedure.In this single centre study we retrospectively analyzed data of 69 large volume leukapheresis (LVL) from 36 low weight (< 20Kg) paediatric patients between January 1998 and April 2008. Primary diagnosis included several malignancies: neuroblastomas (21 cases), meduloblastomas (4 cases), acute leukaemia (4 cases), retinoblastoma (2 cases) and other 5 cases with various solid tumours. All the LVL procedures underwent with a continuous flow blood separator (COBE Spectra) after priming the extracorporeal line with red blood cells and acid-citrate-dextrose (ACD) was used as the anticoagulant.Mean body weight was 15,78kg (ranging 9–19Kg), and a median age of 44 months (ranging 7–88 months). Blood withdrawn at a mean rate of 33,8 mL/min (standard deviation [SD] 8,0), through central vein catheters and using 4 volemias (one case with 3), lasting a mean 133,43 minutes/procedure (ranging 212–101 min). A mean of 4881,78 ml of blood was processed (ranging 2101–7327mL).Leukapheresis was performed on the fifth day of G-CSF stimulation in most of the procedures (66,7%), while 21 patients were submitted to stimulation for a longer period (18,8% with 6 days, 7,2% with 7 days and 4,3% with eight). The mean G-CSF dose used was 203,06 μg/d (ranging 105–400 μg/d).The mean number of CD34+ cells in peripherical blood, before leukapheresis, was 20,4 ×106 cells (ranging 1–216 ×106 cells), while the PBPC collection yielded 2,73 ×106 CD34+/Kg (ranging 0,1–28,94 ×106 CD34+/Kg).The majority of LVL procedures (53,6%) were initiated with a peripherical blood CD34+ count inferior to 15×106 cells. Only 33,3% of each LVL procedure harvest sufficient PBPC (> 2,5 ×106 CD34+/Kg), most of which from patients with peripherical blood CD34+ count superior to 15×106 cells (59,4%). There is a significant correlation between peripherical blood CD34+ count, before leukapheresis, and collected PBPC CD34+ (Pearson chi-square 18,212; p<0,01).Every patient underwent a mean of 1,9 apheresis procedures (range 1–5), 20 patients required only a single apheresis to collect the minimum requirement of CD34+/Kg, 6 patients underwent leukapheresis on two consecutive days, while 7 patients required a second course of mobilization with G-CSF in the following month, to achieve the necessary amount of CD34+ cells.The leucocyte count variation during the procedure was + 8,12 cells/μL (18,37 [SD 9,0]–26,49 cells/μL [SD 28,6]); the platelet (plt) count variation was negative 63,78 ×103 plt/μL (145 [SD 78,5]–81,53×103 plt/μL [SD 39,47]); the haemoglobin count variation was negative 1,45 g/dL (9,79 [SD 1,35]–8,34 g/dL [SD 1,49]). There were no significant differences between heart rate (105,86 [SD 22,9]–113,31 bpm [SD 22,5]) and systolic blood pressure before and after the procedure (114,28 [SD 12,54]–108,38 mmHg [SD 15,72]).There were adverse events in 14,5% of LVL procedures, most of them related with symptomatic hypocalcaemia (6 cases), corrected with calcium injection. Two cases of catheter malfunctioning were reported, one of them associated with haemodinamic instability. There was also a case of high blood pressure peak with no haemodinamic instability and another case with vomiting. All these patients underwent PBPC collection, in the following days.Twenty five percent of the cases needed transfusion support, with 9 cases requiring blood transfusion, and 8 cases requiring platelet transfusion (platelet count fell below 50×103 plt/μL).Nineteen patients underwent autologous transplantation, using the collected cells. There were no incidents regarding the storage of the PBPC.In conclusion, harvesting PBPC in small children is effective and safe, as this data shows. Future publications will allow to gather the necessary experience in order to optimize this procedure.

PBPC collection techniques: standard versus large volume leukapheresis (LVL) in donors and in patients

Transplantations of autologous and allogeneic peripheral blood progenitor cells (PBPC) are able to assure a complete hematopoietic and immunologic reconstitution in patients. PBPC are collected by leukapheresis technique after prior mobilization therapy, but procedures and results remain still highly variable and are poorly characterized. An optimum regimen for PBPC collections has not yet been recommended, but 2–3 total blood volumes (TBV) of the donor or patient are regarded as a standard. Another promising technique is large volume leukapheresis (LVL) with processing of 3–6 TBV of donor or patient. The aim of this paper is to find the most efficient and safe collection technique for an individual donor or patient and, consequently minimize the number of procedures required. Finding the optimal collection procedure would be helpful while considering which method would be preferred in an individual donor or patient with respect to the result of mobilization, health state and required yield of CD 34+ cells for transplantation. We evaluated the results in a total of 134 standard and LVL procedures, which were performed in 21 well mobilized donors (Group I), in 65 well mobilized patients (Group II), and in 14 weakly mobilized patients (Group III) with hemato-oncological diseases. A precollection concentration of CD 34+ cells in peripheral blood higher than 20 × 10 3/mL was considered to be the criterion for efficient mobilization. Such levels of concentration indicating the start of PBPC collections could be easily reached in Group I of donors and Group II of well mobilized patients. Heavily pretreated patients at advanced stages of disease (Group III) did not respond to mobilization sufficiently and had a concentration of CD 34+ cells lower than 20 × 10 3/mL. LVL technique made it possible to obtain higher numbers of CD 34+ cells than in the standard collection in well mobilized donors (Group I), well mobilized patients (Group II), and even in weakly mobilized patients in Group III. In donors and well mobilized patients (Group I and Group II) it was possible to collect sufficient amounts of CD 34+ cells for allogeneic or for autologous transplantation from one LVL collection. The median yield of CD 34+ cells from one LVL collection was 5.5 × 10 6/kg b.w. in donors, and 6.0 × 10 6/kg b.w. in well mobilized patients. Due to the linear dependence of the yield of collected CD 34+ cells on the concentration of CD 34+ cells in blood, it can be used as a simple prediction of the success of collection in Group II (correlation coefficient 0.93 for standard procedures, and correlation coefficient 0.88 for LVL). In Group III of weakly mobilized patients the standard collections were usually ineffective and the relationship between the yield of CD 34+ cells/kg in the product and the precollection concentration of CD 34+ cells was much less significant (correlation coefficient 0.56 for standard procedures and correlation coefficient 0.66 for LVL). The median of CD 34+ cells collected from one standard procedure was only 0.7 × 10 6/kg but using LVL the median increased to 1.4 × 10 6/kg. Our results prove that the yield of CD 34+ cells in the product can be enhanced by large volume leukapheresis (LVL). Based on the results obtained, we recommend LVL in all donors and patients who can tolerate it due to a greater chance of collecting higher yields of progenitor cells while minimizing adverse reactions. LVL procedures should also be preferred in weakly mobilized patients where it is not possible to collect sufficient amounts of CD 34+ cells for transplantation using the standard regime. In weakly mobilized patients LVL provides a greater chance to at least collect a minimum amount of CD 34+ cells necessary. LVL should be used in circumstances where extremely high doses of CD 34+ cells has to be prepared, e.g. planned “tandem” transplantations or manipulations with a graft in which a significant loss of cells is expected.

Hematopoietic progenitor cell large volume leukapheresis (LVL) on the Fenwal Amicus blood separator.

A technique for large volume leukapheresis (LVL) for hematopoietic progenitor cell (HPC) collection using the Fenwal Amicus is presented. It was compared to standard collections (STD) with regard to CD34+ cell yields and cross-cellular content. Optimal cycle volumes and machine settings were evaluated for LVL procedures. A total of 68 patients underwent 80 HPC collection procedures. Because of differences in CD34+ cell yields associated with peripheral white blood cell counts (WBC), the comparison was divided into groups of 20 with WBC < or =35 x 10(9)/L (< or =35 K) and those >35 x 10(9)/L (>35 K). Baseline CD34+ cell counts (peripheral count when patient started HPC collection) were used (median 18-23 cells/microl). Significantly more whole blood (corrected for anticoagulant) was processed with LVL (LVL 20 l vs. STD 13.5 l). For < or =35 K, median CD34+ x 10(6), WBC x 10(9), RBC ml, Plt x 10(11) yields/collection were 183, 21.2, 14, 0.8, respectively, for STD vs. 307, 22.1, 11, 1.0, respectively, for LVL. For >35 K, median CD34+ x 10(6), WBC x 10(9), RBC ml, Plt x 10(11) yields/collection were 189, 32.7, 15, 1.4, respectively, for STD vs. 69, 40.8, 21, 1.3, respectively, for LVL. We have described a method of LVL using the Amicus that, in patients with pre-procedure WBC < or =35 x 10(9)/L, collects more CD34+ cells than a standard procedure with acceptable cross-cellular content. This method is not recommended when pre-procedure WBC counts are >35 x 10(9)/L.

Collection of More Hematopoietic Progenitor Cells with Large Volume Leukapheresis in Patients with Multiple Myeloma

Reinfusion of mobilized peripheral blood stem cells (PBSC) after high dose chemotherapy accelerates hematopoietic recovery. Because of the relatively low content of hematopoietic progenitors in the peripheral blood even after mobilization, multiple leukapheresis procedures are necessary to reach the required target number of CD34 cells to ensure prompt engraftment post-transplantation. Our previous studies have shown that the highest proportions of hematopoietic progenitors cells (CD34) are collected during the first three days of apheresis, whereas peak levels of myeloma cells are observed during subsequent days. Therefore, large volume leukapheresis (LVL), defined as processing of greater than 3 blood volumes or a total of at least 15 liters, was explored in 23 myeloma patients, undergoing 91 procedures; 14 patients were mobilized with high dose cyclophosphamide (6g/m2) and hematopoietic growth factors and 9 with G-CSF only. CD34 yields were measured separately for the first and last two hours of collection. We observed no decrease in CD34 cells/kg during the last two hours of collection and when the LVL collections were compared to historical matched controls, mobilized with the same regimen, the median quantity of CD34 cells/kg/liter collected remained equivalent during all days of apheresis. When compared to G-CSF only, mobilization with high dose cyclophosphamide appeared to result in superior hematopoietic stem cell collections. Interestingly, the G-CSF group experienced a progressive decrease in platelets during consecutive days of LVL, while the opposite was seen in the cyclophosphamide group. LVL procedures were not associated with a higher complication rate than standard volume apheresis. We conclude that LVL procedures allow collection of more CD34 cell per session while not jeopardizing progenitor cell collections during subsequent sessions. Since more CD34 cells are collected, fewer days are required to attain the optimal target of progenitor cells. This should result in PBSC grafts with less tumor contamination.

Engraftment with peripheral blood stem cells collected by large-volume leukapheresis for patients with lymphoma.

Seven patients with refractory lymphomas underwent marrow reconstitution with peripheral blood stem cells (PBSCs) harvested by large-volume leukapheresis (LVL). PBSCs were collected from all patients more than 1 month after the last cycle of chemotherapy, and no patient received growth factors. The median number of LVL procedures performed per patient was 4.5, with a mean volume of 24.5 L of blood processed per procedure to obtain 7 x 10(8) mononuclear cells per kg. Autologous PBSCs and platelets were frozen at a controlled rate in plasma and 10-percent dimethyl sulfoxide and stored in the vapor phase of liquid nitrogen. This group of patients was compared to a control group (n = 18) who received medullary marrow (MM) transplants for the same diagnoses under the same protocols during the same period. Posttransplant days to white cell engraftment (PBSC = 17, MM = 15.5) were no different. Days to platelet independence were significantly longer in the LVL PBSC group (PBSC = 33, MM = 16; p < 0.05). This pattern of engraftment is typical of patients treated in this manner. Although Day 0 platelet counts (PBSC = 75.5 x 10(9)/L, MM = 85 x 10(9)/L) and total single-donor unit platelet use (PBSC = 8, MM = 9) were no different, Day 1 platelet counts (PBSC = 128 x 10(9)/L, MM = 61.5 x 10(9)/L; p < 0.05) and Day 14 platelet use (PBSC = 5, MM = 8; p < 0.05) were significantly different, because of the transfusion of cryopreserved autologous platelets with PBSCs on Day 0.