Prospective flow cytometric evaluation of nucleated red blood cells in cord blood units and relationship with nucleated and CD34(+) cell quantification

BACKGROUND: Cord blood (CB) represents an alternate source of stem cells in transplantation. Nucleated red blood cells (NRBCs) are a physiological subset of CB population. Although it is important to have an accurate estimate of CD34(+) cell number, NRBCs could compromise white blood cell count and interfere with CD34(+) cell quantification. STUDY DESIGN AND METHODS: A total of 826 CB units were analyzed for total nucleated cells (TNCs), NRBCs, and CD34(+) cells by flow cytometry. NRBCs were also counted conventionally by manual microscopy. Percentages of CD34(+) cells corrected by NRBC count (CD34+c) were determined as follows: %CD34+c = CD34(+)/CD45(+) (x10(6))/(TNCs (x10(8)) - NRBCs (x10(8))). RESULTS: The mean percentages of CD34+ cells and NRBCs were 0.27 percent (range, 0.01%-1.25%) and 7.64 percent (range, 0.13%-84%), respectively. Comparison between flow cytometric and microscopic NRBC count showed a regression of y = 0.685 + 0.719x and a coefficient of determination of r(2) = 0.721. When corrected with NRBC count, the mean percentage of CD34(+) c cells was 0.295 percent (p = 0.0008 compared with CD34(+)%) and mean TNCc count was 14.8 x 10(8) (p < 10(-4) compared to TNC count). CONCLUSION: The determination of NRBCs with a flow cytometric method might represent a new strategy for providing satisfactory quality assurance controls of CB products.     

Quantification of nucleated red blood cells in allogeneic marrow graft and impact of processing on recovery

BACKGROUND: In allogeneic bone marrow transplantation (BMT), a higher nucleated and CD34+ cell dose has been reported to improve various outcomes. Other cell types, such as lymphocyte subsets, also influenced BMT results. While nucleated red blood cells (NRBCs) represent a subset of bone marrow (BM) cell subpopulation, the question of their quantification in BM grafts and the impact of BM processing on their recovery has not been addressed. STUDY DESIGN AND METHODS: In a prospective study on 77 BM products, NRBCs were enumerated by flow cytometry and the recovery analyzed after manipulation. Because NRBCs could compromise white blood cell count, the impact of NRBC count on CD34+ cell percentage and total nucleated cell (TNC) dose were also determined. RESULTS: The mean percentage of NRBCs in BM grafts was 21.6 percent (range, 7.8%-40.9%). Mean NRBC recoveries after BM concentration or RBC depletion were 98.4 and 28.7 percent, respectively, close to those obtained for TNC cells (88.6 and 31.3%, respectively). When corrected with NRBC count, the mean percentages of corrected CD34+ cell and TNC dose were significantly modified when compared with uncorrected values, whatever the type of BM manipulation. CONCLUSION: Our data show that NRBC quantification might be of importance to improve quality control of BM products and to evaluate the influence of NRBCs cell dose on outcomes after BMT.     

Impact of donor- and collection-related variables on product quality in ex utero cord blood banking

BACKGROUND: Optimizing product quality is a current focus in cord blood banking. This study evaluates the role of selected donor- and collection-related variables. STUDY DESIGN AND METHODS: Retrospective review was performed of cord blood units (CBUs) collected ex utero between February 1, 2000, and February 28, 2002. Preprocessing volume and total nucleated cell (TNC) counts and postprocessing CD34 cell counts were used as product quality indicators. RESULTS: Of 2084 CBUs, volume determinations and TNC counts were performed on 1628 and CD34+ counts on 1124 CBUs. Mean volume and TNC and CD34+ counts were 85.2 mL, 118.9 x 10(7), and 5.2 x 10(6), respectively. In univariate analysis, placental weight of greater than 500 g and meconium in amniotic fluid correlated with better volume and TNC and CD34+ counts. Greater than 40 weeks' gestation predicted enhanced volume and TNC count. Cesarean section, two- versus one-person collection, and not greater than 5 minutes between placental delivery and collection produced superior volume. Increased TNC count was also seen in Caucasian women, primigravidae, female newborns, and collection duration of more than 5 minutes. A time between delivery of newborn and placenta of not greater than 10 minutes predicted better volume and CD34+ count. By regression analysis, collection within not greater than 5 minutes of placental delivery produced superior volume and TNC count. CONCLUSION: Donor selection and collection technique modifications may improve product quality. TNC count appears to be more affected by different variables than CD34+ count.     

Cesarean section due to fetal distress increases the number of stem cells in umbilical cord blood

BACKGROUND: Umbilical cord blood (UCB) can be used as hematopoietic stem cell source for transplantation. The success of a transplantation is highly correlated with the number of total nucleated cells (TNCs) and CD34+ cells in the UCB. Certain obstetric factors increase the yield of stem cells in the UCB. It is necessary to evaluate optimal conditions in labor to decrease the rate of sample rejection due to low cell count. No data exist regarding the difference between primary and secondary Cesarean sections in terms of efficacy of stem cell harvesting. STUDY DESIGN AND METHODS: Seventy-nine consecutive UCB units from women who had a Cesarean section between 1997 and 2003 were included. The number of TNCs, CD34+ cells, colony-forming units (CFUs), white blood cells (WBCs), nucleated red blood cells (NRBCs), and the total collection volume were compared between cases with primary and secondary Cesarean section. RESULTS: UCB obtained after a Cesarean section due to fetal distress has significantly higher numbers of TNCs, CD34+ cells, NRBCs, and WBCs compared to elective Cesarean section. Of the cases with secondary Cesarean section due to fetal distress, 67 percent resulted in UCB units with sufficient TNC numbers (> or =80 x 10(7) TNCs) compared to 42 percent of the cases with primary Cesarean section. CONCLUSION: Fetal distress increases the number of hematopoietic stem cells mobilized into UCB. Particular effort should be made to collect UCB from newborns who experienced fetal distress.     

Role of large unstained cells in predicting successful stem cell collection in autologous stem cell transplantation

IntroductionSufficient stem cell collection is mandatory for Autologous stem cell transplantation (ASCT). Peripheral CD34+ stem CD34 + stem cell counting by flow cytometry is the gold standard method in both the predicting and timing of successful stem cell collection. Large unstained cells (LUC) are large peroxidase-negative cells that are displayed on certain automatic cell counters and present large lymphocytes, virocytes, blasts, abnormal cells and hematopoietic stem cells. In this study, we evaluated the role of LUC parameters in the timing and prediction of successful stem cell collection.MethodsPatients with a diagnosis of multiple myeloma, lymphoma and testis tumor who proceed to ASCT were included in this study. Preapheresis LUC parameters were analyzed with Siemens ADVIA® 2120i system., Kruskal Wallis, Mann-Whitney U, Spearman Rho and receiver–operator curve (ROC) tests were used for analyses.ResultsNinety patients were evaluated. Peripheral CD34 + cell count was positively correlated with both LUC count (p = 0014) and LUC percentage (p = 0,01). LUC percentage in peripheral blood was positively correlated with mobilized stem cell count in the yield (p = 0.003). We found a LUC count of > 0.485 × 10⁹/L as a cut-off value for detecting > 20 × 10⁶/L CD34 +cells in the peripheral blood with a sensitivity of 64.6% and specificity of 75%. We defined > 2.15% as a cut-off value for LUC percentage to collect > 5 × 10⁶/kg of stem cells with a sensitivity of 64% and specificity of 63%. Additionally, total nucleated cell (TNC) count was negatively correlated with LUC percentage (p = 0.014) and positively correlated with LUC count (p = 0.001).ConclusionLUC parameters are readily available, simple and cheap tools that can be useful in both timing of CD34 count by flow cytometry in peripheral blood and in the prediction of successful mobilization. LUCs can also be an indicator of graft composition.     

Total nucleated cell counts are driving clinician's choice rather than cryopreservation period: Lesson for cord blood banks

According to the increase in both the number of cryopreserved cord blood (CB) units and the cryopreservation period for each CB unit in the largest public CB bank in Korea, we are pursuing greater efficiency in CB bank management. Thus, we analyzed whether the cryopreservation period has a negative impact on the selection of CB units for CB transplantation (CBT). Until December 2019, 468 CB units were used for transplantation. The cryopreservation period, total nucleated cell (TNC), and CD34+ cell counts were analyzed among the CB units according to the CBT-year and the donation year. The results showed that the cryopreservation period was increased in recent CBT-year groups. The transplanted CB units showed similar TNC counts irrespective of the donation year, and the mean TNC count was 13.9 × 10⁸/unit. CB units cryopreserved for a relatively long period were transplanted consistently. The mean TNC count of CB units cryopreserved for over 10 years was 16.4 × 10⁸/unit. The mean CD34+ cell counts were not significantly different among the CB units transplanted after CBT-2013 and among the CB units donated after CBT-2011. Through an analysis of the CB units selected by clinicians for CBT, this study revealed that clinicians placed more weight on the TNC counts than on the cryopreservation period of cryopreserved CB units. Therefore, the minimum TNC count of CB units suitable for cryopreservation should be increased up to 13.0 × 10⁸/unit to balance the satisfaction of clinicians’ needs with the efficiency of the CB bank.     

Effects of cell concentration,time of fresh storage,and cryopreservation on peripheral blood stem cells: PBSC fresh storage and cryopreservation

IntroductionPeripheral blood stem cells are widely used in autologous or allogeneic transplantation. The quality of the product directly impacts clinical outcomes, and the cell quality and/or functionality may be influenced by the storage conditions as time, temperature, total nucleated cells (TNC) concentration and cryopreservation requirement.ObjectiveTo verify the effects of time, cell concentration, and cryopreservation/thawing in the viability and functionality of stem cells for transplantation.MethodsWe evaluated TNC, CD45⁺ viable cells, CD34⁺ viable cells, and cell viability and functionality of 11 samples. Measurements were performed immediately and 24 h, 48 h, 72 h, and 96 h after sample collection at high and low TNC concentrations. The same parameters were also evaluated after cryopreservation and thawing of the samples.ResultDuration of storage and TNC concentration exhibited a negative effect on cell quality (CD45⁺ viable cells, CD34⁺ viable cells and functionality). Moreover, the association of these parameters increased the negative effect on graft quality. Cryopreservation and thawing also negatively affected the collected sample regarding viable CD34⁺ cells (recovery 66.2 %), viable CD45⁺ cells (recovery 56.8 %), and 7-AAD viability. No significant losses in viable CD45⁺/CD34⁺ cells and functionality were observed in the first 24 h in both TNC conditions.ConclusionThese results emphasize the importance to consider carefully the storage conditions until transplantation, measuring TNC/μL until 24 h after collection (diluting the product when TNC > 300 × 10³/μL) and infusing fresh graft as soon as possible.     

The clearance time of infused hematopoietic stem cell from the blood circulation

There is no detailed information about the clearance time of infused hematopoietic stem cell (HSC) from the blood circulation in humans. In this prospective study, peripheral blood CD34+ cell counts were detected during the 4 days period following autologous HSC transplantation in 20 patients by means of flow cytometry. The median CD34+ cells were at the highest level in the first hour and decreased below pre-infusion values on the first day after HSC infusion. By nonparametric analysis, positive correlation was found between CD34+ cell levels at the first hour and the post-thaw CD34+ cell dose (r = 0.57, p = 0.01). An inverse correlation was determined between CD34+ cell levels at the first hour and neutrophil engraftment (r = ?0.54, p = 0.01). Compared with the patients having CD34+ cell count of ?2 μL^?1 in the first hour following HSC infusion, the patients having CD34+ cell count of <2 μL^?1 had delayed both neutrophil (20 vs. 12, p = 0.008) and platelet (47 vs. 11, p = 0.01) engraftments. Our results indicated that infused HSCs were removed from the blood circulation within 1 day. In addition, CD34+ cell levels at the first hour may be used as an important indicator to predict the delay of neutrophil and platelet engraftments.     

Increased numbers of total nucleated and CD34+ cells in blood group O cord blood: an analysis of neonatal innate factors in the Korean population

BACKGROUND: We analyzed neonatal factors that could affect hematopoietic variables of cord blood (CB) donated from Korean neonates. STUDY DESIGN AND METHODS: The numbers of total nucleated cells (TNCs), CD34+ cells, and CD34+ cells/TNCs of CB in neonates were compared according to sex, gestational age, birth weight, birth weight centile for gestational age, and ABO blood group. RESULTS: With 11,098 CB units analyzed, blood group O CB showed an increased number of TNCs, CD34+ cells, and CD34+ cells/TNCs compared with other blood groups. Although TNC counts were lower in males, no difference in the number of CD34+ cells was demonstrated because the number of CD34+ cells/TNCs was higher in males. An increase in the gestational age resulted in an increase in the number of TNCs and decreases in the number of CD34+ cells and CD34+ cells/TNCs. The numbers of TNCs, CD34+ cells, and CD34+ cells/TNCs increased according to increased birth weight centile as well as birth weight. CONCLUSION: CB with blood group O has unique hematologic variables in this large‐scale analysis of Korean neonates, although the impact on the storage policies of CB banks or the clinical outcome of transplantation remains to be determined.     

Quantifying loss of CD34+ cells collected by apheresis after processing for freezing and post-thaw

Introduction: CD34⁺ cells collected for autologous bone marrow transplantation (BMT) are usually quantified in the apheresis product after collection, but the necessity to repeat these measures post-thaw is controversial.Methods: We examined the loss of CD34⁺ cells after collection, preparation for freezing and post-thaw in apheresis products collected for BMT.Results: Median number of CD34⁺ cells collected per unit was 1.61 × 10⁶/kg, viability: 97–100%. This number decreased to 1.38 × 10⁶/kg, viability: 96–100% before freezing and was 1.17 × 10⁶/kg post-thaw. Viability decreased to 86–98%. The relative loss of viable PBHPC showed an inverse correlation with the ratio “CD34⁺ cells/total nucleated cells” (r = ?0.45; p = <0.0005). This relative loss was largest in patients with Hodgkin’s lymphoma.Conclusion: Cryopreservation and thawing of PBHPCs in leukapheresis products provokes a small but significant stem cell loss. So, quantification of viable CD34⁺ cells post-thaw is important, especially in poorly mobilizing patients. Besides, the ratio “CD34⁺ cells/total nucleated cells” after leukapheresis is an important parameter for prediction of neutrophil recovery after BMT.     

Storage characteristics of cord blood progenitor cells: report of a multicenter study by the cellular therapies team of the Biomedical Excellence for Safer Transfusion (BEST) Collaborative

BACKGROUND: Most hematopoietic progenitor cell (HPC) products are infused or processed shortly after collection, but in some cases this may be delayed for up to 48 hours. A number of variables such as temperature and cell concentration are of critical importance for the integrity of HPCs during this time. STUDY DESIGN AND METHODS: We evaluated critical variables using cord blood HPC units that were divided equally and stored at 4°C versus room temperature (RT) for up to 96 hours. Total nucleated cell (TNC) and mononuclear cell (MNC) counts, viable CD34+ cell counts, and CD45+ cell viability as well as colony‐forming unit–granulocyte‐macrophage (CFU‐GM) present over time at each temperature were determined. RESULTS: Overall, the data indicate that with the exception of viable CD34+ cells, there was a significant decrease in each variable measured for 72 to 96 hours and, with the exception of viable CD34+ cells and CFU‐GM, the reductions were significantly greater in RT units than 4°C units. There was an increase in viable CD34+ count for units where TNC count was greater than 8.5 × 10⁹/L, compared with units where TNC count was less than 8.5 × 10⁹/L, that was different for each storage temperature. CONCLUSIONS: Cord blood HPC collections maintained at 4°C retained higher TNC counts, MNC counts, and CD45+ cell viability over a 72‐ to 96‐hour storage period.     

脐血干细胞库的建立及其临床应用

目的建立无关供者脐血干细胞库,用于治疗骨髓衰竭、恶性及非恶性血液病、某些遗传病、重型免疫缺陷等疾病的干细胞移植.方法采集足月顺产或剖宫产的新生儿脐血,以羟乙基淀粉分离红细胞,DMSO/LMD冷冻保存.每份脐血常规检测细胞活率、有核细胞计数、祖细胞体外培养、CD34+细胞检测、HLA配型及病原学检测等.结果共采集2318份脐血,合格1240份,合格率为53.5%,合格脐血采集的平均体积(98.3±27.0)ml,分离后脐血有核细胞为(1.2±0.5)×109,回收率为(88.90±9.65)%.脐血细胞存活率为(97.00±2.61)%.CFU-GM数为(5.18±14.35)×105,BFU-E数为(10.35±11.55)×105,CFU-Mix数为(1.03±1.47)×105.脐血分离后CD34+细胞百分率为(0.5±0.32)%,细菌培养阳性4例,4例脐血CMV-IgM阳性.在广州脐血库的172例脐血移植登记患者中,HLA匹配4个位点以上的占91.12%.至2000年5月共提供9份应用于无关供者脐血移植,其中6例植入,植入成功率66.7%.结论脐血库的建立可使脐血能够较好地应用于造血干细胞移植治疗,建立脐血库有良好的应用前景.     

Impact of apheresis automation on procedure quality and predictability of CD34+ cell yield

Success of peripheral blood stem cell (PBSC) collections depends on patient biological parameters and stable apheresis device performance. We investigated product quality and factors influencing main apheresis procedure outcomes including CD34⁺ collection efficiency (CE), product volume or platelet CE. We also assessed different CD34⁺ cell yield prediction algorithms. Autologous PBSC collections by Spectra Optia from myeloma and lymphoma patients were analyzed. Complete blood count (CBC) from patient preprocedure and from collected products were assessed. (1) Product yield was calculated, (2) Product CBC was correlated with patient preprocedure variables, and (3) Predictions of CD34⁺ yields based on (a) product CD34⁺ cell concentration in samples after two or four chamber flushes or (b) traditional CE2 benchmark, were compared. 62 procedures in 41 patients were analyzed. 84% of all procedures were run without operator intervention. Median CD34⁺ CE2 was 56.9% (48.8%‐65.2%) and quite stable irrespective of patient conditions, with minor influence from patient white blood cell (WBC) precounts (r_s = –.47; P < .001). Platelet loss correlated with WBC precount (r_s = .46; P < .001), product volume (r_s = .71; P < .0001) and number of chambers collected (r_s = .72; P < .0001). CD34⁺ cell yield was better predicted based on (a) product CD34⁺ cell concentration from samples after 2 and 4 chamber flushes, respectively (r_s = .969; P < .0001 and r_s = .9648; P < .0001) than based on (b) CE2 formula (r_s = .8262, P < .0001). Spectra Optia provides good quality PBSC products with stable and predictable yield regardless of starting conditions. CD34⁺ sampling of product after few chamber flushes could be used to predict CD34⁺ yield.     

Enhanced mobilization of CD34<Superscript>+</Superscript> progenitor cells expressing cell adhesion molecules in patients with STEMI

Background  Adult stem cells can contribute to myocardial regeneration after ischemic injury. The aim of the study was to determine (1) the amount of mobilized CD34⁺/CD117⁺, CD34⁺/KDR⁺ cells into peripheral blood (PB) in relation to inflammatory and haematopoietic cytokines, (2) the presence of circulating CD34⁺ cells, expressing cell adhesion molecules (CAM), in patients with ST-segment elevation myocardial infarction (STEMI) in comparison to patients with coronary artery disease (CAD). Materials and methods  Twenty-three patients with STEMI (<12 h), 24 patients with CAD and 15 control subjects were enrolled in this study. The patients were matched in age, 2-CAD, ejection fraction (45%) and end-diastolic volume index (70 ml/m²). The number of stem cells and the expression of adhesion molecules were quantified by use of flow cytometry. Inflammatory cytokines [interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF), vascular endothelial growth factor] and chemotactic factors as stromal cell-derived factor-1 (SDF-1), hepatocyte growth factor (HGF) were determined by ELISA. Results  The amount of circulating progenitor cells including CD34⁺/CD117⁺ and CD34⁺/KDR⁺ cells was significantly higher in patients with STEMI than in patients with CAD (CD34⁺/CD117⁺ 433 ± 128 vs. 100 ± 17, P = 0.012; CD34⁺/KDR⁺ 253 ± 41 vs. 128 ± 24, P = 0.02). The mobilization of CD34⁺ progenitor cells expressing CXCR4-receptor, lymphocyte function-associated antigen-1 (LFA-1), very late antigen-4 (VLA-4) and ICAM-1 into PB was significantly higher in patients with STEMI compared to CAD (CD34⁺/CXCR4⁺ 740 ± 327 vs. 136 ± 23, P = 0.006; CD34⁺/LFA-1 976 ± 227 vs. 329 ± 41, P = 0.025; CD34⁺/VLA4⁺ 830 ± 161 vs. 330 ± 31, P = 0.007; CD34⁺/ICAM⁺ 387 ± 66 vs. 144 ± 26, P < 0.001). Additionally, the cytokines G-CFS, IL-6 and HGF were upregulated and significantly increase in the STEMI group compared with controls and CAD (G-CSF 50.6 ± 6.8 vs. 23 ± 3 vs. 23.8 ± 2, P ^{Co vs. STEMI} < 0.001, P ^{Co vs. CAD} = n.s., P ^{STEMI vs. CAD} < 0.001; IL-6 8.4 ± 0.6 vs. 3.8 ± 1.9 vs. 2.6 ± 1, P ^{Co vs. STEMI} < 0.001, P ^{Co vs. CAD} = n.s., P ^{STEMI vs. CAD} < 0.001; HGF 4,502 ± 461 vs. 686 ± 195 vs. 1,746 ± 461, P ^{Co vs. STEMI} < 0.001, P ^{Co vs. CAD} = n.s., P ^{STEMI vs. CAD} < 0.001), while the level of SDF-1 was increased in patients with CAD compared to controls and patients with STEMI (3,035 ± 286 vs. 2,028 ± 76 vs. 2,154 ± 234, P ^{Co vs. STEMI} = n.s., P ^{Co vs. CAD} = n.s., P ^{STEMI vs. CAD} = 0.005). Conclusions  The study demonstrates in patients with STEMI an increased mobilization of progenitor cells like CD34⁺/CD117⁺ and CD34⁺/KDR⁺ compared to CAD. Furthermore, we could shown that in patients with STEMI the mobilization of CD34⁺ progenitor cells with expressed CAM was increased. It is to speculate that an enhanced expression of adhesion molecules may increase the transmigration and implantation of progenitor cells into ischemic myocardium for myocardial repair.     

Beneficial ex vivo immunomodulatory and clinical effects of clarithromycin in COVID-19

IntroductionMacrolide antibiotics have immunomodulatory properties which may be beneficial in viral infections. However, the precise effects of macrolides on T cell responses to COVID, differences between different macrolides, and synergistic effects with other antibiotics have not been explored.MethodsWe investigated the effect of antibiotics (amoxicillin, azithromycin, clarithromycin, and combined amoxicillin with clarithromycin) on lymphocyte intracellular cytokine levels and monocyte phagocytosis in healthy volunteer PBMCs stimulated ex vivo with SARS-CoV-2 S1+2 spike protein. A retrospective cohort study was performed on intensive care COVID-19 patients.ResultsCo-incubation of clarithromycin with spike protein-stimulated healthy volunteer PBMCs ex vivo resulted in an increase in CD8⁺ (p = 0.004) and CD4⁺ (p = 0.007) IL-2, with a decrease in CD8⁺ (p = 0.032) and CD4⁺ (p = 0.007) IL-10. The addition of amoxicillin to clarithromycin resulted in an increase in CD8⁺ IL-6 (p = 0.010), decrease in CD8⁺ (p = 0.014) and CD4⁺ (p = 0.022) TNF-alpha, and decrease in CD8⁺ IFN-alpha (p = 0.038). Amoxicillin alone had no effect on CD4⁺ or CD8⁺ cytokines. Co-incubation of azithromycin resulted in increased CD8⁺ (p = 0.007) and CD4⁺ (p = 0.011) IL-2. There were no effects on monocyte phagocytosis. 102 COVID-19 ICU patients received antibiotics on hospital admission; 62 (61%) received clarithromycin. Clarithromycin use was associated with reduction in mortality on univariate analysis (p = 0.023), but not following adjustment for confounders (HR = 0.540; p = 0.076).ConclusionsClarithromycin has immunomodulatory properties over and above azithromycin. Amoxicillin in addition to clarithromycin is associated with synergistic ex vivo immunomodulatory properties. The potential benefit of clarithromycin in critically ill patients with COVID-19 and other viral pneumonitis merits further exploration.     

The comparison of Filgrastim (Neupogen®), biosimilar filgrastim (Leucostim®) and Lenograstim (Granocyte®) as a first line peripheral blood stem cell mobilization strategy in autologous hematopoieitic stem cell transplantation: A single center experience from Turkey

Objectives and aimPatients affected by hematological malignancies can often benefit from high dose chemotherapy followed by peripheral blood stem cells (PBSCs) transplantation. Different strategies have been used to mobilize an adequate number of PBSC, including granulocyte colony-stimulating factor (G-CSF) alone or chemotherapy plus G-CSF. In this study, we aimed to compare the efficacy profile of different G-CSF agents including filgrastim (Neupogen®), biosimilar filgrastim (Leucostim®) and Lenograstim (Granocyte®) on CD34⁺ mobilization in patients who underwent autologous hematopoietic stem cell transplantation (autoHSCT).Materials and methodsWe retrospectively analysed data of patients who underwent autoHSCT diagnosed with multiple myeloma (MM), Hodgkin Lymphoma (HL), non-Hodgkin Lymphoma (NHL) and others. Data for stem cell mobilization has been obtained from patients’ files. Patients who received Filgrastim (Neupogen®), biosimilar Filgrastim (Leucostim®, Group) and Lenograstim (Granocyte®) were evaluated mainly for total CD34⁺ cell count at the end of mobilization procedure.ResultsA total of 96 patients who underwent autoHSCT were retrospectively analyzed. 27 (28.2%) of the patients were female, and 69 (71.8%) were male. The diagnosis of the patients were; multiple myeloma (39 patients, 40.6%), Hodgkin Lyphoma (23 patients, 23.9%), non-Hodgkin lymphoma (16 patients, 16.6%), and others (18 patients, 18.9%). The median number of leukapheresis cycle necessary to harvest a minimal count of 3 × 10⁶ CD34⁺/kg was 2 in Neupogen® (min–max: 1–4) and Granocyte® (min–max: 1–3) groups and 1 (min–max: 1–2) in Leucostim® group. The median doses of G-CSF agents (μg/kg/day) in PBSC collection procedure were; 10.00 (min–max: 7.00–12.00) in the Neupogen® group, 8.00 (min–max: 7.25–9.00) in the Leucostim® group and 8.50 (6.00–9.50) in the Granocyte® group. There was no statistical significance among groups (p = 0.067). The number of total collected PB CD34⁺ cells (×10⁶/kg) was 7.64 (min–max: 4.09–13.86) in the Neupogen® group, 13.43 (min–max: 8.15–23.38) in the Leucostim® group and 5.45 (min–max: 4.28–9.40) in the Granocyte® group. The data showed that patients in the leucostim group had significantly higher PB CD34⁺ cells compared to patients in the Granocyte® group (p = 0.013).ConclusionLeucostim® was comparable to Neupogen® for PBSC mobilization in patients who underwent autoHSCT.     

Factors predicting the hematopoietic stem cells content of the umbilical cord blood

Umbilical cord blood (UCB) has been demonstrated to be alternative source of hematopoietic stem cells (HSCs). Unfortunately, the wide use of UCB Transplantation is limited due to the low number of HSCs. The aim of this study was to determine factors that affect the number of HSCs collected from UCB. 200 eligible donors were included for HSCs testing, including total nucleated cells (TNCs) and CD34+ cell number, by using univariate and multivariate analysis. In univariate analysis, factors positively associated with higher number of TNCs were maternal weight (P = 0.002), preeclampsia (P = 0.03), neonatal weight (P < 0.001), neonatal platelet count (P = 0.02), neonatal Rh (P = 0.03), gestational age (P = 0.04) and delivery type (P < 0.001). Factors positively associated with higher number of CD34+ cells were maternal weight (P < 0.007), preeclampsia (P = 0.02), maternal hypertension (P = 0.02) neonatal weight (P < 0.001), neonatal Rh type (P = 0.02) and delivery type (P = 0.04). In multivariate analysis, factors significantly influence TNCs were neonatal weight (P < 0.001), preeclampsia (P = 0.008), neonatal Rh type (P = 0.02) and delivery type (P < 0.001). While factors significantly influence number of CD34+ cells were maternal weight (P = 0.025), neonatal weight (P = 0.005), neonatal Rh (P = 0.006), nuchal cord (P = 0.026) and delivery type (P = 0.009). Conclusions factors significantly influence TNCs content of UCB were neonatal weight, preeclampsia, neonatal Rh and delivery type. While factors significantly influence number of CD34+ cells were maternal weight, neonatal weight, neonatal Rh, nuchal cord and delivery type.     

Comparative Efficacy and Safety of Fostemsavir in Heavily Treatment-Experienced People With HIV-1

PurposeHeavily treatment-experienced (HTE) people with multidrug-resistant HIV-1 have limited treatment options. Treatment with the first-in-class attachment inhibitor fostemsavir in addition to optimized background therapy (OBT) resulted in sustained virologic and immunologic responses in HTE participants throughout 96 weeks in the BRIGHTE trial. In the absence of long-term direct comparative evidence between fostemsavir-based and other antiretroviral regimens, this analysis indirectly compares efficacy and safety across relevant available trials, adjusting for demographic and baseline characteristics.MethodsA systematic literature review was conducted to identify trials with designs and populations comparable to BRIGHTE. Using matching-adjusted indirect comparison analyses, individual participant data from BRIGHTE were reweighted to create balanced populations across trials, and efficacy and safety outcomes were compared.FindingsThree comparator trials were identified, 2 of which reflected an optimized therapy without fostemsavir (OBT alone): TMB-301 (ibalizumab and OBT), BENCHMRK-1/-2 (OBT alone), and VIKING-3 (OBT alone). Compared with ibalizumab and OBT (N = 40), fostemsavir and OBT (unadjusted, N = 347; adjusted, N = 236) were associated with numerically higher nonsignificant odds of virologic suppression (odds ratio [OR] = 1.44; 95% CI, 0.74–2.80; P = 0.284) and a similar increase in CD4⁺ cell count of approximately 65 cells/mm³ from baseline through week 24 (mean difference = 7.05 cells/mm³; 95% CI, ?60.88 to 74.98 cells/mm³; P = 0.834). Compared with OBT from BENCHMRK-1/-2 (N = 237), fostemsavir and OBT (adjusted, N = 126) were associated with significantly higher odds of virologic suppression (OR = 3.26; 95% CI, 2.08–5.11; P < 0.001) and increased CD4⁺ cell count (135.78 cells/mm³; 95% CI, 91.93–179.63 cells/mm³; P < 0.001) at week 96. Compared with OBT from VIKING-3 (N = 183), fostemsavir and OBT (adjusted, N = 78) were associated with numerically higher odds of virologic suppression (OR = 1.34; 95% CI, 0.78–2.30; P = 0.297) and a modest CD4⁺ cell count increase (26.86 cells/mm³; 95% CI, ?10.79 to 64.52; P = 0.162) through week 48; however, differences were not significant. All-cause discontinuations and safety comparisons varied across studies.ImplicationsAlthough matching-adjusted indirect comparison analyses have limitations, these results support the use of fostemsavir and OBT as an important treatment option in HTE people with multidrug-resistant HIV-1.     

The relationship of CD34+ dosage and platelet recovery following high dose chemotherapy and autologous CD34+ reinfusion in multiple myeloma

Autologous hematopoietic stem cell transplantation (ASCT) is an established treatment for multiple myeloma (MM), yet the impact of transplanted CD34+ cell dose remains unresolved, especially in patients over the age of 65 years. Data was collected from 207 consecutive ASCT patients to determine the relationship between CD34+ infusion count and short-term and long-term platelet recovery. For MM patients under the age of 65 years (n = 155), CD34+ dosage correlates with time to platelet engraftment (p < 0.001) and platelet count at 30 days (p = 0.003), but not with long-term platelet counts at 180 or 360 days from the CD34+ reinfusion. For MM patients aged 65 years or older (n = 46), CD34+ dosage did not correlate with time to platelet engraftment, but did correlate with both short-term and long-term platelet counts at 30 (p < 0.001), 180 (p = 0.021), and 360 days (p = 0.005). Exploratory regression analysis was done to explore platelet stability following the current minimum CD34+ dosage reinfusion. For MM patients under the age of 65 years, the minimum standard CD34+ dosage of 2 × 10⁶ cells/kg was sufficient for a timing to platelet engraftment of <21 days and short-term platelets count ≥150 × 10⁹/L at 30 days. Alternatively, for MM patients aged 65 years or older, the CD34+ dosage of 2 × 10⁶ cells/kg was insufficient for platelet counts ≥150 × 10⁹/L at 30 and only marginally attainable at 360 days suggesting that in elderly MM patients a higher CD34+ dosage may be required for platelet recovery and possibly long-term platelet stability.