Collection, cryopreservation and thawing of stem cells for children weighing less than 25 Kg with high-risk neuroblastoma: A single center results in Morocco

ABSTRACT Introduction: An important component of the advances made in neuroblastoma treatment has been the use of peripheral blood stem cells to support high-dose chemotherapy. In this study, we report our experience on a series of small children who have undergone standard and large volume leukaphersis (LVL) procedures, provide an update on a single institution's experience with cryopreservation of autologous peripheral blood stem cells (PBSCs), using 10% dimethyl sulfoxide (DMSO) and applying post-thaw DMSO depletion and analyze a number of variables that may affect viability. Methods: A total of 36 aphereses were performed on 29 children weighing less than 25 kg between July 2016 and October 2019 at the Ibn Sina university hospital. Results: Seven females and twenty-two males, median bodyweight 14 kg (9 - 22). A single apheresis was sufficient to obtain at least 3 × 106/kg body weight (BW) of CD34+ cells in 82.8% of the cases. The LVL was performed in 22 aphereses. A median number of 5.9 × 106/ kg CD34 cells were collected per apheresis. A total of 60 PBSC samples were cryopreserved and 46 samples were infused. The mean cell viability percentage decreased from 94.75 ± 1.14% before freezing to 70.84 ± 8.6% after thawing (p < 0.001). No correlation was found between post-thaw viability and storage time (r = -0.233; p = 0.234) or number of total nucleated cells (r = 0.344; p = 0.073). Conclusion: Leukapheresis is safe and feasible in small pediatric patients if the appropriate measures are used. Cryopreservation poses numerous challenges, especially a decrease in cell viability after thawing.

A Single Center 25-year Experience in Autologous Peripheral Blood Stem Cell Collection: A Focus on the Collection Efficiency

Background:Harvest of hematopoietic progenitor cells via leukapheresis is being used increasingly for autologous transplantation. Adequate yield of cells per kilogram body weight of recipient is required for a successful engraftment. Collection efficiency(CE)is a useful parameter to assess quality of peripheral blood stem cell (PBSC) collection program. In this study, we report a 25-year experience in a tertiary care Hospital in Italy.Patients and Methods:1,026 consecutives autologous PBSC collection procedure, performed in 763 patients, from January 1996 to December 2020 were retrospectively considered.Data regarding patients, Blood Cells Separators (BCS) , apheresisprocedures and PBSC products were collected in our database. In these 25 years different BCSwere adopted in our Apheresis Unit (AU). In the first period (1996-1999) we used Fresenius Com.Tec, in the central period (2000-2013) we used Cobe Spectra and inthe last period (2014-2020) Spectra Optia. Results:As regards the evaluation of patients before leukapheresis, the most significant data was the increasing number of CD34+ cells. Considering the PBSC collection procedure, there was a progressive increase in the processed blood volume with a shorter apheresis duration. Data related to the PBSC collection demonstrated an increasing CD34+ cell yield and efficiency a raise in CE that was 43% using Fresenius COM-TEC BCS, 49% using Cobe Spectra BCS and 53% using Spectra Optia BCS. . Conclusions:These results were observed considering a 25-year period, thus a great number of factors likely contributing to the observed results, including technological improvement of the instrumentation for leukapheresis,increased experience of the team operating in the Apheresis Unit, improved mobilization protocols, better criteria for patients’ selection. Focusing our attention on CE we observed quite satisfactory results with a median which rose from 43% to 53% with an increase of 10% in the observation period.

Effects of Leukapheresis on Hemostatic Function in Patients with Hyperleukocytic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To analyze and compare the effects of leukapheresis on hemostatic function in patients with hyperleukocytic leukemia.@*METHODS@#A total of 139 patients with AML, ALL and CML who underwent leukapheresis from June 2009 to February 2020 and did coagulation test before and after operation were included in this study. The clearance efficiency of each group and the difference among three groups were evaluated, as well as hemostatic function including platelet counts, coagulation indicators, CDSS score and incidence of adverse events. The difference of hemostatic function caused by leukapheresis in different leukemia patients were compared.@*RESULTS@#After leukapheresis, the WBC counts were decreased significantly in the three groups of patients (P<0.001), and the clearance efficiency was highest in ALL patients. However, the platelet counts also were decreased significantly (AML:P＜0.001, ALL: P＜0.001, CML: P＜0.01) in the three groups of patients, particularly for acute leukemia patients with a positive correlation with WBC clearance efficiency(r=0.284). After leukapheresis, fibrinogen decreased, PT and APTT prolonged. For acute leukemia patients, higher CDSS score was related to an elevated incidence of bleeding events (P<0.05).@*CONCLUSION@#Leukapheresis is an effective method to decrease the leukemic burden, but it is necessary to monitor the impact on hemostatic function. It is recommended to assess the CDSS socre for acute leukemia patients, in order to identify the predictive value for bleedings.

Prospective identification of potential factors influencing stem cell mobilization and the necessity for plerixafor use in newly diagnosed multiple myeloma patients undergoing autologous stem cell transplantation

ABSTRACT Introduction: To study the efficacy and safety of single large volume leukapheresis by using generic G-CSF or G-CSF plus Plerixafor in achieving adequate stem cell yield and various factors influencing thereof in newly diagnosed multiple myeloma patients undergoing autologous stem cell transplant . Method: This prospective study was undertaken among 55 newly diagnosed multiple myeloma patients undergoing autologous stem cell transplant and aged between 18 and 75 years. Mobilization and harvesting of stem cells were performed by using GCSF or GCSF plus Plerixafor and large volume leukapheresis, respectively. A stem cell yield of ≥2 × 106 kg-1 and the number of apheresis procedures were primary efficacy endpoints, while the ideal stem cells yield >5 × 106 kg-1, the engraftment day and D100 response/graft sustainability were secondary endpoints. Result: The primary endpoint was achieved in all cases in both the groups by using a single LVL leukapheresis procedure. Fulfillment of all the secondary endpoints was satisfactory and comparable in both the groups. Age, pre-apheresis CD34+ count and number of interruptions during the LVL were significant factors influencing the stem cell yield (p < 0.05). Adverse drug reactions during the apheresis and post-ASCT period were manageable. Conclusion: The LVL is safe and cost-effective in attaining a minimum of CD34+ cells in a single procedure with manageable adverse reactions. Judicious intervention during the procedure may be helpful in ensuring the adequate yield.

Validation of a formula predictive of peripheral blood stem cell yield and successful collection in healthy allogeneic donors

ABSTRACT Background: An efficient mobilization and collection of peripheral blood stem cells (PBSCs) are crucial to optimize engraftment in the recipient. We aim to validate a formula that predicted CD34+ cell yield and to describe variables that correlated with high yield mobilization and collection in healthy donors. Methods: We retrospectively analyzed clinical and laboratory data from healthy donors who underwent PBSC collection from 2006 to 2015. The predicted number of collected cells was calculated using the following formula: Total number of CD34+ (cells × 106/kg) yield = [(peripheral CD34+ cells/µL) × (0.43)/recipient body weight (kg)] × total liters processed. Results: We evaluated 338 collections from 307 allogeneic PBSC donors. The predicted versus the observed number of CD34+ cells/kg collected yielded an r-value of 0.775 (0.726-0.816; p < 0.0001). Overall, 55.7% donors had an acceptable mobilization level. Donors with a body weight <67 kg were less likely to yield a satisfactory CD34+ cell count (OR = 0.44; 95% CI 0.24-0.81), while a white blood cell (WBC) count >40 × 109/L (OR = 3.69; 2.11-6.46) and platelet count ≥200 × 109/L (OR = 2.09; 1.26-3.47) on the day of collection predicted a good level of mobilization. Predictors of a CD34+ cell yield/kg of ≥4 × 106 with only one apheresis session were: circulating CD34+ cells/µL >40 (OR = 16; 6.94-36.93), hemoglobin ≥14 g/dL (OR = 3.40; 1.53-7.57), WBC >40 × 109/L (OR = 4.61; 2.10-10.10) on the first collection day, and a positive delta weight between donor and recipient (OR = 3.10; 1.36-7.06). Conclusion: The formula for predicting CD34+ cell yield is accurate and suggests the optimal length of time for successful leukapheresis. Validation of the predictors of successful mobilization will help to further refine PBSC leukapheresis procedures.

CD34+ hematopoietic progenitor cells enumeration and mononuclear cell count: an experience from a tertiary care centre

Background: This study was conducted to assess the CD34+ hematopoietic progenitor cells enumeration by flow cytometry and the utility of performing mononuclear cell count before performing the Stem cell enumeration. Collection was done on two consecutive days with CD34+ hematopoietic progenitor cell enumeration of both the samples. Mononuclear cell counts were done in all the patients. The purpose of the study was to do counts directly from the leukapheresis pack and see the reliability of this practice.Methods: Samples were collected from the leukapheresis pack and subjected to mononuclear cell count and CD34+ hematopoietic progenitor cells enumeration by flow cytometty before harvesting.Results: A total of 66 samples from 34 patients were taken up for the study. 76.47% of our cases were that of multiple myeloma and 17.64% of the cases were that of non Hodgkin lymphoma and 2.94% cases each of neuroblastoma and Hodgkin lymphoma. It was noted that the mononuclear cell counts correlated well with the CD34+ HPC in most of the cases with MNC being above 4 x 108 per pack per kg body weight in cases where CD34+ HPC counts were more than the desired lower limit of 2 x 106 per pack per kg body weight.Conclusions: It was observed that flow cytometric enumeration of CD34+ hematopoietic progenitor cells directly from the leukapheresis pack gave satisfactory results even without doing peripheral blood CD34+ HPCs enumeration before leukapheresis. Also, in our study we were able to set a limit of mononuclear Cell at 4 x 108 per pack/kg BW as counts beyond that always correlated with the more accurate flow cytometric method of CD34+ HPC count of more than 2 x106 per pack/kg body weight, therefore acting as a crude method for assessing the mobilization.

Efficacy of Granulocyte and Monocyte Adsorptive Apheresis for Treatment of Inflammatory Bowel Disease / 胃肠病学

The prevalence of inflammatory bowel disease (IBD) in China is increasing year by year, however, the efficacy and safety of commonly used therapeutic methods are limited.Granulocyte and monocyte adsorptive apheresis (GMA) is one of the effective methods for treatment of IBD used abroad, however, there is still lacking of such research in China.Aims: To investigate the efficacy and safety of GMA in IBD patients.Methods: A retrospective study was conducted in 21 cases of IBD patients [13 cases with ulcerative colitis (UC) and 8 with Crohn's disease (CD)] who accepted GMA treatment from May 2013 to July 2014 at the Shanghai Rui Jin Hospital.All the cases were poor responders to 5-aminosalycylic acid (5-ASA) or steroid-refractory.The clinical data were collected, and the clinical activity index (CAI), endoscopic activity index (EAI), laboratory parameters including serum albumin (Alb), hemoglobin (Hb), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), leukocyte count and percentage of neutrophils, as well as the adverse effects before and two weeks after the end of GMA treatment were analyzed.Results: After GMA treatment, both CAI and EAI were decreased significantly in UC and CD groups as compared with those before treatment (P all 0.05).The treatment was well tolerated with no severe adverse effects.Conclusions: GMA is safe and effective for ameliorating clinical symptoms, attenuating intestinal mucosal injury and controlling active inflammation in IBD patient that has not responded to 5-ASA or steroid treatment.Prospective clinical studies with large samples are needed to confirm these findings.

Therapeutic Leukapheresis for the Management of Hyperleukocytosis: Ten-year Experience in a Tertiary Care Hospital

BACKGROUND: Hyperleukocytosis is a medical emergency that is characterized by increased blood viscosity and predisposition to various neurological, pulmonary, and gastrointestinal complications. In addition, patients are at risk of the tumor lysis syndrome because of the increased tumor burden. Therapeutic leukapheresis is an important treatment for these emergent states. In this study, we retrospectively analyzed therapeutic leukapheresis procedures that were performed in our institution during the last 10 yr. METHODS: We retrospectively analyzed therapeutic leukapheresis procedures conducted from July 2005 to March 2015 at a tertiary care hospital. We present our observations, especially the procedural characteristics and hematological parameters before and after the aforementioned procedures. RESULTS: Seventy-two patients underwent a total of 146 therapeutic leukapheresis procedures. The average presenting white blood cell (WBC) count was 268×10(3)/µL, and ranged from 54×10(3)/µL to 673×10(3)/µL. After an average of two sessions, a statistically significant drop in the WBC counts was observed. The average WBC removal rates during the initial and entire therapeutic leukapheresis procedures of each patient were 33% and 46%, respectively. The platelet count and hemoglobin concentration were significantly reduced. CONCLUSIONS: Therapeutic leukapheresis significantly reduces peripheral WBC counts and is a safe and effective procedure for the treatment of hyperleukocytosis.

Mobilization and collection of CD34+ cells for autologous transplantation of peripheral blood hematopoietic progenitor cells in children: analysis of two different granulocyte-colony stimulating factor doses

The use of peripheral hematopoietic progenitor cells (HPCs) is the cell choice in autologous transplantation. The classic dose of granulocyte-colony stimulating factor (G- CSF) for mobilization is a single daily dose of 10 µg/kg of patient body weight. There is a theory that higher doses of granulocyte-colony stimulating factor applied twice daily could increase the number of CD34+ cells collected in fewer leukapheresis procedures. Objective: The aim of this study was to compare a fractionated dose of 15 µg G-CSF/kg of body weight and the conventional dose of granulocyte-colony stimulating factor in respect to the number of leukapheresis procedures required to achieve a minimum collection of 3 × 106 CD34+ cells/kg body weight. Methods: Patients were divided into two groups: Group 10 - patients who received a single daily dose of 10 µg G-CSF/kg body weight and Group 15 - patients who received a fractioned dose of 15 µg G-CSF/kg body weight daily. The leukapheresis procedure was carried out in an automated cell separator. The autologous transplantation was carried out when a minimum number of 3 × 106 CD34+ cells/kg body weight was achieved. Results: Group 10 comprised 39 patients and Group 15 comprised 26 patients. A total of 146 apheresis procedures were performed: 110 (75.3%) for Group 10 and 36 (24.7%) for Group 15. For Group 10, a median of three (range: 1-7) leukapheresis procedures and a mean of 8.89 × 106 CD34+ cells/kg body weight (±9.59) were collected whereas for Group 15 the corresponding values were one (range: 1-3) and 5.29 × 106 cells/kg body weight (±4.95). A statistically significant difference was found in relation to the number of apheresis procedures (p-value <0.0001). Conclusions: To collect a minimum target of 3 × 106 CD34+ cells/kg body weight, the administration of a fractionated dose of 15 µg G-CSF/kg body weight significantly decreased the number of leukapheresis procedures performed.

Clinical Effects and Safety of Therapeutic Leukapheresis in Hyperleukocytosis / 대한수혈학회지

BACKGROUND: Therapeutic leukapheresis is the cytoreduction procedure performed before chemotherapy in patients with hyperleukocytosis for prevention of complication. However, there have been clinical concerns about bleeding tendency due to anticoagulant used during the procedure. The aim of our study was to compare the clinical characteristics and hematological parameters before and after therapeutic leukapheresis in order to evaluate its effect on bleeding tendency and to provide a guideline for treatment strategy. METHODS: The clinical data for 39 procedures of therapeutic leukapheresis performed on 17 patients with hyperleukocytosis from May 2005 to October 2013 at the National Cancer Center were reviewed retrospectively. RESULTS: The patients consisted of 11 males and six females. The mean age was 41 years old (range, 8~74). The mean number of therapeutic leukapheresis per patient was two (range, 1~4). Clinical symptoms improved in 14 patients (82%) after therapeutic leukapheresis and three patients (18%) were not yet to improve. The mean WBC count was significantly reduced by 32.6% (+/-17.4) after therapeutic leukapheresis, from 250,146/microL (+/-117,000) to 174,702/microL (+/-104,700) (P<0.001). The mean volume of single removal was 298 ml with 4.25x10(11)/L (+/-1.54) WBCs. After therapeutic leukapheresis, the mean platelet count showed a decline from 85x10(9)/L (+/-43) to 71x10(9)/L (+/-26). However, the prothrombin time (PT) and activated partial thromboplastin time (aPTT) did not show a significant increase (PT, P=0.637; aPTT, P=0.054). CONCLUSION: Therapeutic leukapheresis is demonstrated as an effective and safe treatment that can improve symptoms and reduce leukocytes in hyperleukocytosis.

Effect of leukocyte-depleted intraoperative salvaged blood on erythrocyte immunity and systemic inflammatory response during perioperative period / 中华麻醉学杂志

Objective To investigate the effect of leukocyte-depleted intraoperative salvaged blood on the erythrocyte immunity and systemic inflammatory response during the perioperative period in patients.Methods Twenty ASA Ⅰ or Ⅱ patients (aged 20-44 years and weighing 40-75 kg) requiring blood salvage during operation (with the estimated intraoperative blood loss of 15-20 ml/kg) were randomly divided into two groups (n =10 each):control group (group C) and leukocyte depletion group (group D).Anesthesia was induced with midazolam,fentanyl,propofol and vecuronium and maintained with isoflurane inhalation and intravenous infusion of propofol and remifentanil.The patients were mechanically ventilated after endotracheal intubation.Intraoperative blood salvage and reinfusion were performed in all the patients.In group D,the salvaged blood was filtered by a leukocyte depletion filter placed in the line of the reinfusion circuit.Blood samples were collected from the central vein before anesthesia (T1),at the end of surgery (T2) and 12 hours (T3) and 36 hours (T4) after operation in all the patients.The rosette rates of red blood cell-C3b receptors (RBC-C3bRR) and RBC-immune complex (RBC-ICR) were determined.The number of leukocytes and polymorphonuclear neutrophils (PMNs) were counted.The plasma levels of interieukin-6 (IL-6),tumor necrosis factor-alpha (TNF-α) and malondialdehyde (MDA) were measured,too.Results Compared with group C,the plasma levels of IL-6,TNF-α and MDA were significantly lower at T2-T4,the RBC-C3bRR was significantly higher at T3-T4 (P ＜ 0.01),and the number of PMNs was significantly lower at T4 in group D(P＜0.05).Conclusion Leukocyte-depleted intraoperative salvaged blood is helpful to improve the erythrocyte immunity during the perioperative period in patients,and the decrease of systemic inflammatory response may be involved in the mechanism.

Effects of leukocyte-depleted allogeneic blood transfusion on perioperative cellular immunity in children / 中华麻醉学杂志

Objective To evaluate the effects of leukocyte-depleted allogeneic blood transfusion on perioperative cellular immunity in children.Methods Three hundred and fifty-nine ASA Ⅰ or Ⅱ children (aged 3 months-14 years and weighing 5-74 kg) requiring allogeneic blood transfusion during operation were randomly divided into two groups:163 children receiving normal allogeneic blood transfusion (control group,group C) and 196 children receiving leukocyte-depleted allogeneic blood transfusion (group D).Blood samples were collected from the peripheral vein before blood transfusion,and 2 and 6 days after blood transfusion for determination of the levels of CD3+,CD4+,CD8 +,and CD56+ by flow cytometry.CD4+ /CD8+ ratio was calculated.The volume of allogeneic blood transfusion during operation,the duration of operation,postoperative drainage,antibiotic administration,hospital stay and the incidence of postoperative infection were recorded.Rssults The levels of CD3+,CD4+,CD56+ and CD4+/CD8+ ratio significantly increased at 6 days after blood transfusion while the duration of postoperative drainage,postoperative antibiotic administration,hospital stay and the incidence of postoperative infection significantly decreased in group D compared with group C.Conclusion Leukocyte-depleted allogeneic blood transfusion is helpful in improving the postoperative cellular immunity in children.

Effect of leukocyte-depleted intraoperative salvaged blood on erythrocyte immunity and systemic inflammatory response during perioperative period / 中国医师进修杂志

Objective To investigate the effect of leukocyte-depleted intraoperative savaged blood on erythrocyte immunity and systemic inflammatory response during perioperative period patients.Methods Sixty patients required blood salvage were randomly divided into two groups by random digits table method with 30 cases each.The patients in control group were given routine autologous blood transfusion,while in observation group,the salvaged blood was filtered with a leukocyte depleting filter placed in the reinfusion circuit.Blood samples were collected from the central vein before anesthesia (T1),at the end of surgery(T2),and at 12 h (T3) and 36 h (T4) after operation in two groups.The rosette rates of RBC-C3b receptors (RBC-C3bRR) and RBC-immune complex (RBC-ICR) were determined.The leukocyte and neutrophil were counted.The plasma levels of interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α) and malondialdehyde (MDA) were measured.Results Compared with control group,the plasma level of IL-6,TNF-α,MDA at T2-T4 in observation group was decreased[T2:(17.8 ± 1.6) ng/L vs.(20.8 ± 1.3) ng/L,(17.4 ± 2.3) ng/L vs.(23.1 ± 2.1) ng/L,(4.31 ± 0.33) nmol/L vs.(4.64 ± 0.73) nmol/L;T3:(21.2 ± 2.0) ng/L vs.(24.3 ± 2.4) ng/L,(12.3 ±2.1) ng/L vs.(18.6 ±1.9) ng/L,(3.97 ±0.35) nmol/L vs.(4.43 ±0.64) nmol/L;T4:(22.0 ± 1.3) ng/L vs.(28.4 ± 1.5) ng/L,(10.6 ± 1.7) ng/L vs.(14.6 ± 2.2) ng/L,(3.45 ± 0.57) nmol/L vs.(3.95 ± 0.40) nmol/L],RBC-C3bRR at T3-T4 in observation group was increased [T3:(15.3 ± 1.3)/100 RBC vs.(12.8 ± 1.5)/100 RBC ;T4:(15.8 ± 1.2)/100 RBC vs.(13.0 ± 1.5)/100 RBC],and there were significant differences (P ＜0.05).There was no significant difference in RBC-ICR,the leukocyte and neutrophil between two groups (P ＞ 0.05).Conclusion Leukocyte-deleted intraoperative salvaged blood is helpful to improve the erythrocyte immunity during perioperative period in patients,and the decrease in the systemic inflammatory response may be involved in the mechanism.

Fatores que afetam a coleta de células-tronco hematopoiéticas do sangue periférico por leucaférese de grande volume: experiência de um único centro / Factors affecting autologous peripheral blood hematopoietic stem cell collections by large-volume leukapheresis: a single center experience

Objective: To evaluate factors affecting peripheral blood hematopoietic stem cell yield in patients undergoing large-volume leukapheresis for autologous peripheral blood stem cell collection. Methods: Data from 304 consecutive autologous peripheral blood stem cell donors mobilized with hematopoietic growth factor (usually G-CSF), associated or not with chemotherapy, at Hospital Israelita Albert Einstein between February 1999 and June 2010 were retrospectively analyzed. The objective was to obtain at least 2 x 106 CD34+ cells/kg of body weight. Pre-mobilization factors analyzed included patient?s age, gender and diagnosis. Post mobilization parameters evaluated were pre-apheresis peripheral white blood cell count, immature circulating cell count, mononuclear cell count, peripheral blood CD34+ cell count, platelet count, and hemoglobin level. The effect of pre and post-mobilization factors on hematopoietic stem cell collection yield was investigated using logistic regression analysis (univariate and multivariate approaches). Results: Premobilization factors correlating to poor CD34+ cell yield in univariate analysis were acute myeloid leukemia (p = 0.017) and other hematological diseases (p = 0.023). Significant post-mobilization factors included peripheral blood immature circulating cells (p = 0.001), granulocytes (p = 0.002), hemoglobin level (p = 0.016), and CD34+ cell concentration (p < 0.001) in the first harvesting day. However, according to multivariate analysis, peripheral blood CD34+ cell content (p < 0.001) was the only independentfactor that significantly correlated to poor hematopoietic stem cell yield. Conclusion: In this study, peripheral blood CD34+ cell concentration was the only factor significantly correlated to yield in patients submitted to for autologous collection.

Objetivo: Avaliar fatores que afetam o rendimento da coleta em pacientes submetidos à leucaférese de grande volume para obtenção de células-tronco hematopoiéticas do sangue periférico para autotransplante. Métodos: Análise retrospectiva de 304 doadores de células-tronco hematopoiéticas de sangue periférico para autotransplante submetidos à mobilização com fator de crescimento hematopoiético (geralmente G-CSF), associado ou não à quimioterapia, no Hospital Israelita Albert Einstein de Fevereiro de 1999 a Junho de 2010. O objetivo da coleta foi obter pelo menos 2x106 CD34+ células/kg peso. Os fatores pré-mobilização incluíam idade, sexo e diagnóstico do paciente. Os parâmetros pós-mobilização avaliados foram contagem de leucócitos, células imaturas, células mononucleares e células CD34+, plaquetas e nível de hemoglobina no sangue periférico. O efeito desses fatores no rendimento da coleta de CTH foi investigado por meio de regressão logística (análise univariada e multivariada). Resultados: A análise univariada revelou os seguintes fatores pré-mobilização estatisticamente significantes: diagnóstico de leucemia mieloide aguda (p = 0,017) e outras doenças hematológicas (p = 0,023), células imaturas circulantes (p = 0,001), granulócitos (p = 0,002), nível de hemoglobina (p = 0,016) e contagem de células CD34+ (p <0,001) no primeiro dia de coleta. Entretanto, só a contagem de células CD34+ no sangue periférico manteve-se associada de forma significante ao rendimento ruim da coleta de células-tronco hematopoiéticas na análise multivariada. Conclusão: Neste estudo, a contagem de células CD34+ no sangue periférico foi o único fator significantemente associado ao rendimento da coleta de células-tronco hematopoiéticas com leucaférese de grande volume para autotransplante.

Changes of coagulation and lung injury in response to leukocytapheresis in endotoxemia dog / 中华创伤杂志

Objective To observe the effect of leukocytapheresis(LCAP)on the coagulation,fibrinolysis system and lung injury in the endotoxemia dog and explore the mechanism in the endotoxin-induced lung injury dog. Methods Endotoxemia-induced model in dogs was established by administration of lipopolysaccharide(LPS,2 mg/kg).Separation of the leucocytes wag performed with the automated continuous flow blood cell separators.A total of 30 male mongrel dogs were randomly divided into LPS group(group L,only injected with LPS,with no LCAP),sham LCAP group(group S,received sham LCAP at 12-14 hours after administration of LPS)and LCAP treatment group(group T,received LCAP at 12-14 hours after administration of LPS),10 dogs per group.The dynamic changes of the activated protein C(APC),soluble thrombomodulin and plagminogen activator inhibitor-1 in the serum were measured at 0 hour before LPS administration,at 2,6,12,14,16,24 and 36 hours after administration of LPS.Results Through LCAP,there found the following four results:(1) the APC level in the serum of the group T wag(50.805±4.422)μg/ml and(40.480±2.993)μg/ml at 14 hours and 16 hours respectively,which were significantly higher than(45.881±4.024)μml and(35.935±4.057)μg/ml in the group L(P<0.05).(2)The expressions of soluble thrombomadulin in the group T was (9.688±O.914)μml and(10.492±O.865)μg/ml at 14 hours and 16 hours respectively,which was statistically lower than(11.005±0.854)μg/ml and(12.04±0.954)ug/ml in the group L(P<0.05).(3)Thelevel of plagminogen activatorinhibitor-1 in the group T was lower than that in the group the group T Wag statistically lower than that in the group L(ALI/ARDS occurred in 2 and 7 dogs of the groups T and L respectively within 36 hours after infusion of LPS.P<0.05). Conclusions At the decrease the incidence of acute lung injury partly due to its role in improving the function of coagulation and fibrinolysis.

Effect of leukocyte-depleted intraoperative salvaged Mood on erythrocyte immunity and systemic inflammatory response during perioperative period / 中华麻醉学杂志

Objective To investigate the effect of leukocyte-depleted intraoperative salvaged blood on the erythrocyte immunity and systemic inflammatory response during the perioperative period in patients. Methods Twenty ASA Ⅰ or Ⅱ patients, aged 20-44 yr, weighing 40-75 kg, required blood salvage during operations (intraoperative blood loss expected 15-20 ml/kg), were randomly divided into 2 groups ( n = 10 each): control group (group C) and leukocyte depletion group (group D) . Anesthesia was induced with midazolam, fentanyl, propofol and vecuronium and maintained with isoflurane inhalation and iv infusion of propofol and remifentanil. The patients were mechanically ventilated after tracheal intubation. Intraoperative blood salvage and reinfusion were performed in all patients using cell saver system. In group D, the salvaged blood was filtered with a leukocyte depleting filter placed in the line of the reinfusion circuit. Blood samples were collected from the central vein before anesthesia (T1 ) , at the end of surgery (T2 ) , and at 12 h (T3 ) and 36 h (T4 ) after operation in the two groups. The rosette rates of RBC-C3b receptors and RBC-immune complex were determined. The leukocyte and neutrophil were counted. The plasma levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and malondialdehyde (MDA) were measured. Results The plasma levels of IL-6, TNF-a and MDA were significantly lower at T2-4 , the rosette rate of RBC-C3b receptors was significantly higher at T3-4 ( P ＜ 0.01) , and the neutrophil count was significantly lower at T4 in group D than in group C ( P ＜ 0.05) . Conclusion Leukocyte-depleted intraoperative salvaged blood is helpful to improve the erythrocyte immunity during the perioperative period in patients, and the decrease in the systemic inflammatory response may be involved in the mechanism.

Establishment of a off-pump leukocyte depletion method in canines / 中华麻醉学杂志

Objective To establish a off-pump leukocyte depletion method in canines.Methods Twenty-one healthy adult mongrel dogs of both sexes, weighing 10-12 kg, were randomized into 2 groups: control group (group C, n = 9) and leukocyte depletion group (group LD, n = 12) . An extracorporeal leukocyte filtration end was constructed by aseptic puncture of the bilateral external jugular veins via a blood transfusion line and the leukocyte filter for extracorporeal leukocyte depletion. In group LD, blood was filtered with a MP-300 blood line and a SQ40S leucocyte depletion filter one end placed in the right external jugular vein (artery end) and the other end placed in the left external jugular vein (venous end). After heparin anticoagulation, a MP-300 blood line pump was used as the power. Blood was filtered at a rate of 75 ml/min and it was maintained for 60 min. The artery end was then closed, normal saline injected into the closed circuit, the remaining blood pumped into the body, and then the venous end closed. In group C, aseptic puncture of the bilateral external jugular veins was performed. Arterial blood samples were taken immediately before leukodepletion (baseline, T_0 ) , at 10, 20, 30, 40, 50 and 60 min of leukodepletion (T_(1-6) , depletion period), and at 10, 20, 30, 40, 50, 60, 120, 180 and 270 min after leukodepletion (T_(7-15) , recovery period) for determination of blood routine. MAP, HR, RBC.Plt and body temperature were recorded at T_0 , T_6 , T_(12) and T_(15) . Results There were no significant difference in MAP and RBC between the two groups ( P > 0.05 ) . HR, body temperature, and Plt were significantly lower in group LD than in group C (P < 0.05) .The leukocyte concentration was lower during depletion period in group LD than in group C ( P < 0.05) , while there was no significant difference in leukocyte concentration during recovery period between the two groups (P > 0.05 ). Conclusion The off-pump leukocyte depletion method is successfully established and has exact efficacy with less adverse effects in this study.

Cytapheresis in Patients with Severe Ulcerative Colitis after Failure of Intravenous Corticosteroid: A Long-Term Retrospective Cohort Study

BACKGROUND/AIMS: Cytapheresis (CAP) is a novel strategy for ulcerative colitis (UC). However, there is insufficient data on the long-term outcome of UC patients who achieve remission by CAP. This study involved patients with severe UC who refracted to intravenous (iv) corticosteroid. METHODS: Forty-seven UC patients who had received CAP therapy for the first time within 1 year after UC diagnosis were followed for 36 months. One of the inclusion criteria was a clinical activity index (CAI) of > or =7 points at the end of a 2-week iv course of corticosteroid therapy. CAP therapy consisted of ten sessions over 10 weeks. RESULTS: CAP induced clinical remission (CAI or =12, n=25) than for moderately severe UC at entry (7< or =CAI<12, p=15; p<0.02). The cumulative rates of avoiding surgery and relapse were 54.5% and 24.2%, respectively, at 36 months in patients who responded to CAP therapy. This was similar to that of iv cyclosporine reported recently. CONCLUSIONS: This study suggest that CAP is an effective therapy in patients who are refractory to conventional medications including iv corticosteroid. Increased remission rates should be expected in refractory patients with moderately severe UC.

Effect of Leukapheresis on Gene Expression Profiles of Donor's Peripheral Blood Mononuclear Cells / 대한진단검사의학회지

BACKGROUND: Leukapheresis has commonly been used to obtain the cell products intended for clinical cell therapy. Hypocalcemia related to citrate toxicity and some circulatory effects such as hypovolemia and hypotension are well-known complications of leukapheresis. In this study, we analyzed the gene expression profiles of peripheral blood mononuclear cells (PBMCs) obtained before and after leukapheresis to determine if the hemodynamic changes can affect the gene expression profiles of leukocytes. METHODS: PBMCs were isolated from EDTA blood from 5 healthy donors collected before and immediately after apheresis. RNA was isolated, amplified, and analyzed using a cDNA microarray with 17,500 genes. Hierarchical clustering analysis was performed to evaluate the differences of gene expression profiling. RESULTS: Hierarchical clustering separated PBMCs from different donors with each other, but did not separate PBMCs collected before and after leukapheresis. Comparison of gene expression by PBMCs collected before and after leukapheresis found only 25 genes were differentially expressed (15 were up-regulated and 10 were down-regulated after leukapheresis) (F-test, P<0.005). Stress induced apoptosis-related genes, ANXA3, DEDD, and ATXN2L, and cytokine-related genes, IL13RA1 and IK, which were also related to stress, were up-regulated after leukapheresis. Genes involved in DNA and protein binding, such as CLSTN3, LRBA, SATB2, and HSPA8, were down-regulated. CONCLUSIONS: Leukapheresis had little effect on gene expression of PBMCs. Some genes showing differences between before and after leukapheresis were mainly involved in stress-related reactions.

The study of performing therapeutic leukapheresis and plateletpheresis sequencially with a single apheresis kit / 中国实用内科杂志

0.05).Conclusion Performing leukapheresis and plateletpheresis sequencially with a single Apheresis Kit is a safe,effective,practicable and economic method in the treatment of chronic myeloid leukemia with leukocytosis and thrombocytosis.It can drop the medical expanse markedly.