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Abstract Respiratory failure, intracranial hemorrhage and infection were more common in hyperleukocytic acute myeloid leukemia patients than in non-hyperleukocytic leukemia patients. Compared with non-apheresis treatment, the white blood cells decreased significantly and the infection rate decreased after apheresis treatment. However, the treatment time of leukapheresis in patients with hyperleukocytic leukemia is very long, while it is more damaging to cells. In this study, which conducted a retrospective analysis on patients with hyperleukocytic acute myeloid leukemia, the process of centrifugation of normal cells and patients’ cells by apheresis machine was simulated in vitro. Through selecting 5 healthy persons and 11 patients with hyperleukocytic acute myeloid leukemia, extracting their blood samples and performing in vitro centrifugation at different speeds or duration, we observed the changes of the numbers and morphology of peripheral blood cells in healthy people and patients, so as to explore the optimal centrifugation parameters during leukapheresis. The cells obtained by the optimal centrifugation parameters were cryopreserved and two groups of mice (10 mice in each group) were used to establish leukemia animal models. Through the research, it is found that when the centrifugal speed is below 6000 rpm, the damage to blood cells in healthy people and in patients with hyperleukocytic leukemia is not obvious. When the centrifugal speed is above 6000 rpm, the platelets will be damaged significantly. The cells obtained under the optimal centrifugation parameters can be successfully cryopreserved and used to establish leukemia animal models. This study is of great significance for improving the efficiency and reducing the side effects of leukapheresis, and is helpful to improve the treatment of white blood cells reduction.

Hyperleukocytosis is a rare and potentially life-threatening condition that can cause leukostasis and organ dysfunction. This case report describes the case of a 69-year-old female who presented with fever, drowsiness, and consolidation in the left lung lower lobe. Initial investigations showed an increased total leukocyte count (TLC) of 89 × 103/μL, which further increased to 111.9 × 103/μL despite treating with intravenous antibiotics. A bone marrow biopsy revealed myeloid hyperplasia, but the BCR-ABL fusion gene test was negative. Due to symptomatic leukostasis and rapid deterioration of the patient’s condition, therapeutic leukapheresis (TL) was performed. During the first sitting, the device parameters were optimized for maximum platelets and the leukocytes removal. The second sitting of TL was carried out on the next day and the TLC was further reduced to 22.3 × 103/μL. The patient’s general condition and mental status improved significantly and she was taken off the mechanical ventilation and discharged after 7 days with a TLC count of 14.59 × 103/μL. She was regularly followed up for the next 3 months and the counts have remained within normal limits. The case reports highlight that TL may be helpful in similar clinical conditions with symptomatic leukostasis.

INTRODUCTION: Granulocyte transfusion is one of the best therapeutic modalities in prolonged neutropenic patients with severe bacterial/fungal infections. Granulocyte harvest using conventional acid citrate dextrose (ACD) anticoagulant (ACD-A) by apheresis is not satisfactory in comparison to the use of hydroxyethyl starch (HES), but the latter is associated with various adverse events, especially with high-molecular-weight HES. AIMS AND OBJECTIVE: This study aimed to assess the beneficial impact of the use of medium-molecular-weight (MMW)-HES and trisodium citrate combination over ACD-A in granulocyte apheresis when using Spectra Optia. MATERIALS AND METHODS: This was a retrospective study comparing granulocyte harvest results with the use of ACD or HES and trisodium citrate combination. All the donors in both the groups received single 600 μg of granulocyte colony-stimulating factor subcutaneous injection followed by 8 mg of dexamethasone tablet 10–12 h and omnacortil 60 mg orally 3 h before harvest. A number of adverse incidents, if any, were observed and noted. Donor/procedure parameters were compared using Mann–Whitney U-test/unpaired t-test. RESULTS: Granulocyte yield (mean: 3.29 × 1010/unit vs. 4.5 × 1010/unit in the ACD and HES groups, respectively, P ≤ 0.0001) was significantly better in the HES group. The collection efficiency was also better in the HES group (mean: 15.86% vs. 26.70% in the ACD and HES groups, respectively, P ≤ 0.0001) in the ACD and HES groups, respectively. There was no significant adverse event noted in any of these two groups. CONCLUSION: In our study, granulocytes with optimum yield can be easily harvested with Spectra Optia cell separator using 6% HES (MMW) and trisodium citrate combination with standard 12-h interval gap between mobilization and harvest. This strategy can also have no or minimal extra cost burden to patients.

Abstract The majority of adoptive cellular therapies are produced from peripheral mononuclear cells obtained via leukapheresis and further enriched for the cells of interest (e.g., T cells). Here, we present a first‐of‐its‐kind closed system, which effectively removes ~85% of monocytes and ~88% of platelets, while recovering ~88% of concentrated T cells in a separate output stream, as the leukapheresis sample flows through a microfluidic device at 5 mL/min. The system is driven by a common peristaltic pump, enabled by a novel pressure wave dampener, and operates in a closed bag‐to‐bag configuration, without requiring any specialized, dedicated equipment. When compared to standard density gradient centrifugation on paired samples, the new system demonstrated a 1.5‐fold increase in T cell recovery and a 2‐fold reduction in inter‐sample variability for this separation outcome. The T cell‐to‐monocyte ratio of the leukapheresis sample was increased to 20:1, whereas with density gradient processing it decreased to 2:1. As a result of superior purity and/or gentler processing, T cells enriched by the system showed a 2.7‐times higher fold expansion during subsequent culture, and an overall 3.5‐times higher cumulative yield. This centrifugation‐free and label‐free closed system for enriching lymphocytes could significantly simplify and standardize the manufacturing of life‐saving cellular therapies.

Autologous chimeric antigen receptor-modified T-cell (CAR T) products have demonstrated un-precedent efficacy in treating many relapsed/refractory B-cell and plasma cell malignancies, leading to multiple commercial products now in routine clinical use. These positive responses to CAR T therapy have spurred biotech and big pharma companies to evaluate innovative production methods to increase patient access while maintaining adequate quality control and profitability. Autologous cellular therapies are, by definition, manufactured as single patient batches, and demand has soared for manufacturing facilities compliant with current Good Manufacturing Practice (cGMP) regulations. The use of a centralized production model is straining finite resources even in developed countries in North America and the European Union, and patient access is not feasible for most of the developing world. The idea of having a more uniform availability of these cell therapy products promoted the concept of point-of-care (POC) manufacturing or decentralized in-house production. While this strategy can potentially decrease the cost of manufacturing, the challenge comes in maintaining the same quality as currently available centrally manufactured products due to the lack of standardized manufacturing techniques amongst institutions. However, academic medical institutions and biotech companies alike have forged ahead innovating and adopting new technologies to launch clinical trials of CAR T products produced exclusively in-house. Here we discuss POC production of CAR T products.

Leukapheresis is a resource-intensive and high-risk treatment with unclear benefits when used for leukostasis syndrome in hematologic malignancies. In this case report and literature review we discuss the pathophysiology of leukostasis syndrome associated with chronic lymphocytic leukemia (CLL) and the rapidly evolving paradigm of CLL treatment through the lens of a 51-year-old man who was diagnosed with CLL. He presented with clinical manifestations of leukostasis syndrome with an absolute lymphocyte count of 522.6×109/L. We identified 15 additional cases of CLL-associated leukostasis syndrome in our literature review. Pulmonary and neurologic manifestations of leukostasis were most common. In combination with pharmacologic cytoreduction, leukapheresis was used successfully in most cases with few reports of complications. In the context of greater adoption of first line therapies for CLL that are known to induce transient leukocytosis, we explore leukapheresis as an adjunctive therapy that can rapidly reduce the lymphocyte count and in select instances possibly mitigate the effects of Bruton's tyrosine kinase inhibitor-induced hyperleukocytosis.

Objectives One of the treatment modalities that can be used for hyperleukocytosis is leukapheresis. However, the result of studies showing the benefit of early mortality through the use of leukapheresis versus no leukapheresis is still inconclusive. Hence, we aimed to conduct a systematic review with meta-analysis to determine the effect of leukapheresis on early mortality in AML patients with hyperleukocytosis. Methods We conducted a literature search on five databases (PubMed, EBSCOhost, Scopus, Clinicalkey, and JSTOR) up to October 2021 for studies comparing early mortality outcomes between hyperleukocytosis AML patients treated with leukapheresis versus no leukapheresis. Summary odds ratios (OR) and 95% confidence intervals (CI) were calculated using random-effects models. Heterogeneity tests were presented in I2 value and publication bias was analyzed using a funnel plot. Results Eleven retrospective cohort studies were eligible based on the inclusion and exclusion criteria. Pooled analysis showed that there was no significant difference in early mortality between patients receiving leukapheresis and not receiving leukapheresis in studies using hyperleukocytosis cutoff of 95,000/mm3 or 100,000/mm3 (OR: 1.17; 95% CI: 0.74-1.86; p: 0.50; I2: 0%). Similarly, studies using hyperleukocytosis cutoff of 50,000/mm3 also showed no benefits of early mortality (OR: 0.67; 95% CI: 0.43-1.05; p: 0.08; I2: 0%). Most of the studies used had a moderate risk of bias due to being observational studies. Funnel plot showed an indication of publication bias on studies using hyperleukocytosis cutoff of ≥50,000/mm3. Conclusion The use of leukapheresis does not provide early mortality benefit in adult AML patients with hyperleukocytosis.

Abstract Priapism is a rare presenting feature of chronic myeloid leukemia (CML) in male patients. Treatment aims to relieve symptoms and to prevent erectile dysfunction. Several treatment modalities exist, however no standard treatment is recommended. We evaluated literature concerning different treatment approaches and evaluate the value of leukapheresis in treatment of priapism. The literature search resulted in 57 included articles, consisting of 53 studied patients. Patients had a mean age of 25.3 years, average time from onset to presentation at the hospital was 2 days, and mean white blood cell (WBC) count was 344 × 109/L. Most patients (67.9%) were treated with a combined approach (different modalities were radiological, urological, and oncological treatment). Twelve patients, with a mean WBC count of 365 × 109/L, received leukapheresis. Only two of them reported erectile dysfunction after treatment. Priapism is an urological emergency requiring urgent multidisciplinary treatment. We highlight the importance of local urological therapy combined with systemic therapy for CML. Therapeutic leukapheresis should be applied when available and with no other contraindications.

Abstract Leukemic retinopathy is a severe complication of severe leukocytosis that results from untreated chronic myelogenous leukemia (CML). Immediate cytoreduction via leukapheresis may reverse ocular manifestations and prevent permanent vision damage. We present a case of a patient with acute unilateral vision loss found to have leukemic retinopathy in the setting of untreated CML with improvement of visual symptoms after leukapheresis and initiation of hydroxyurea.

Purpose: We aimed to identify subsets of patients who benefit from emergency LA and to establish a therapeutic algorithm for AML patients with hyperleukocytosis. Methods: In this single-center retrospective cohort study, a total of 20 consecutive patients underwent LA because of their clinical symptoms. Overall survival (OS) analysis was conducted using the Kaplan–Meier plot method. Univariate and multivariate analyses (using multiple logistic regression) were performed. At the time of diagnosis, all patients received a standard diagnostic workup for AML including FLT3-ITD mutational analysis. Results: FLT3-ITD mut AML patients receiving LA had a median OS of 437 days (range 5–2379 days) with a corresponding 14-day survival of 92.3%, while FLT3 wt AML patients displayed a significantly lower median OS of only 5 days (range 1–203 days) with a corresponding 14-day survival of 14.3% (p = 0.0001). Conclusions: Among patients with clinical symptoms of leukostasis, the subset of FLT3-ITD mut AML patients showed a better outcome with lower early mortality after emergency LA. Based on these observations, we established a therapeutic algorithm for AML patients with hyperleukocytosis.

IntroductionLeukapheresis reduces hyperleukocytosis in children with acute leukemia. Although the usefulness of this procedure is under debate, a repeated small-volume exchange transfusion along with leukapheresis yielded satisfactory results.MethodsForty-seven patients with acute leukemia [32 acute lymphocytic leukemia (ALL) and 15 acute myeloblastic leukemia (AML)] were enrolled between January 2017 and June 2022 and underwent repeated small-volume exchange transfusion. The following were measured: demographic and clinical characteristics, time of the procedure, PWBC (peripheral white blood cell) count, hemoglobin, platelet count, blood biochemistry, electrolytes, coagulation, leukostasis, TLS (tumor lysis syndrome), DIC (disseminated intravascular coagulopathy), adverse events (AEs), and serious AEs (SAEs).ResultsThe demographic and clinical characteristics were not significantly different between ALL and AML patients, but differences were observed in PWBC counts (424.2 ± 135.6 vs. 223.8 ± 58.0 × 109/L). The procedures needed 3–8 processes, and the average procedure time was not significantly different between ALL and AML. The PWBC count gradually reduced to

Priapism is a rare presentation of Chronic Myeloid Leukaemia (CML). It is also considered a medical emergency as delay in treatment may lead to impotence. Prompt medical and surgical interventions such as hydroxyurea, analgesia, phenylephrine injection and aspiration, open surgical shunting, and local radiation therapy are essential. Leukapheresis effectively reduces leukocyte count rapidly and effectively, thereby an important therapy along with other standard of care in CML-induced priapism. In the present case, priapism was the presenting symptom of CML. The same was managed with various modalities such as hydroxyurea, allopurinol, antibiotics, analgesics, sedatives, phenylephrine injection, and aspiration but failed to reduce priapism pain. With a single cycle of leukapheresis, priapism pain could be reduced significantly.

Pediatric chronic myeloid leukemia (CML) is a very rare malignancy (age-related incidence 0.1/100,000) typically presenting with leucocyte counts >100,000/µL. However, clinical signs of leukostasis are observed at diagnosis in only approximately 10% of all cases and among these, priapism is infrequent. Here, we analyze data from pediatric CML registries on the occurrence of priapism heralding diagnosis of CML in 16/491 (3.2%) boys (median age 13.5 years, range 4–18) with pediatric CML. In the cohort investigated, duration of priapism resulting in a diagnosis of CML was not reported in 5 patients, and in the remaining 11 patients, occurred as stuttering priapism over 3 months (n = 1), over 6 weeks (n = 1), over 1–2 weeks (n = 2), over several days (n = 2), or 24 h (n = 1), while the remaining 4 boys reported continuous erection lasting over 11–12 h. All patients exhibited splenomegaly and massive leukocytosis (median WBC 470,000/µL, range 236,700–899,000). Interventions to treat priapism were unknown in 5 patients, and in the remaining cohort, comprised intravenous fluids ± heparin (n = 2), penile puncture (n = 5) ± injection of sympathomimetics (n = 4) ± intracavernous shunt operation (n = 1) paralleled by leukocyte-reductive measures. Management without penile puncture by leukapheresis or exchange transfusion was performed in 3 boys. In total, 7 out 15 (47%) long-term survivors (median age 20 years, range 19–25) responded to a questionnaire. All had maintained full erectile function; however, 5/7 had presented with stuttering priapism while in the remaining 2 patients priapism had lasted

Aim:Autologous hematopoietic stem cell transplantation is the most frequently used treatment method in the treatment of lymphoma and myeloma patients. To apply this treatment method, first of all, a sufficient number of stem cells must be collected from the patient. With the development of apheresis methods and safe, effective mobilization methods, it is now possible to collect stem cells in an outpatient manner. In our study, we aimed to compare the efficacy and safety of outpatient based mobilization versus inpatient based mobilization of hematopoietic stem cells with granulocyte-colony stimulating factor (G-CSF) alone in patients with myeloma and lymphoma.Materials and Methods:A total of 89 patients, including 54 patients who underwent outpatient and 35 patients who underwent inpatient based mobilization of stem cells with G-CSF alone were included in the study. Outpatient and inpatient based mobilization groups were compared in terms of efficacy and safety. Statistical analyses were performed with Jamovi 1.2.27 software. The Mann-Whitney U and chi-square tests were used to examine the differences. MANCOVA was used for univariate and multivariate statistical analysis of factors influencing mobilization.Results:Three leukaphereses resulted in the collection of a mean 9.73x106/kg (4.5-16.5) CD34+ cells in the outpatient based mobilization group and a mean 11.8x106/kg (3.56-59) CD34+ cells in the inpatient based mobilization group (p=0.14). Life-threatening side effects were not observed in any of the patients. Grade 1, 2 side effects were observed and there was no significant statistical difference between the two groups.Conclusion:In this study, we found no significant difference in terms of efficacy and safety between the outpatient and the inpatient based mobilization group patients with myeloma and lymphoma who were mobilized with G-CSF. The results of our study show that outpatient based mobilization can be effectively and safely performed with g-csf, especially in patients who need autologous transplantation and avoid hospitalization, as in the current Coronavirus disease-2019 pandemic.

Drug therapy of immune-mediated inflammatory arthropathies is not always satisfactory, and there is a risk of adverse events. Granulocyte and monocyte/macrophage apheresis (GMA) is a non-pharmacological therapeutic option that is beneficial and very well tolerated. GMA involves passing blood through a column with cellulose acetate beads to remove increased and activated myeloid lineage cells and improve the cytokine profile. The technique reduces pain and inflammation. We present four clinical reports that illustrate the clinical uses of GMA with the medical device Adacolumn® in patients with different backgrounds and immune-mediated inflammatory arthritis. The results were positive, and no adverse events were reported.

Difficulties in the collection of hematopoietic stem and progenitor cells (HSPCs) from Fanconi anemia (FA) patients have limited the gene therapy in this disease. We have investigated (ClinicalTrials.gov, NCT02931071) the safety and efficacy of filgrastim and plerixafor for mobilization of HSPCs and collection by leukapheresis in FA patients. Nine of eleven enrolled patients mobilized beyond the threshold level of 5 CD34+ cells/μL required to initiate apheresis. A median of 21.8 CD34+ cells/μL was reached at the peak of mobilization. Significantly, the oldest patients (15 and 16 years old) were the only ones who did not reach that threshold. A median of 4.27 million CD34+ cells/kg was collected in 2 or 3 aphereses. These numbers were markedly decreased to 1.1 million CD34+ cells/kg after immunoselection, probably because of weak expression of the CD34 antigen. However, these numbers were sufficient to facilitate the engraftment of corrected HSPCs in non-conditioned patients. No procedure-associated serious adverse events were observed. Mobilization of CD34+ cells correlated with younger age, higher leukocyte counts and hemoglobin values, lower mean corpuscular volume, and higher proportion of CD34+ cells in bone marrow (BM). All these values offer crucial information for the enrollment of FA patients for gene therapy protocols.

Dongdong Zhang,1 Yufan Zhu,2 Yanxia Jin,3 Natasha Mupeta Kaweme,3 Youhong Dong1 1Department of Oncology, Xiangyang No. 1 People’s Hospital, Hubei University of Medicine, Xiangyang, Hubei, 441000, People’s Republic of China; 2Orthopedics department, Zhongnan Hospital, Wuhan University, Wuhan, People’s Republic of China; 3Department of Hematology, Zhongnan Hospital, Wuhan University, Wuhan, People’s Republic of ChinaCorrespondence: Youhong DongDepartment of Oncology, Xiangyang No. 1 People’s Hospital, Hubei University of Medicine, Xiangyang, Hubei, 441000, People’s Republic of ChinaEmail dongyouhong2005@163.comAbstract: Hyperleukocytosis is a hematologic crisis caused by excessive proliferation of leukemic cells and has a relatively high early mortality due to a series of severe complications. Therefore, prompt and effective intervention is required. Leukapheresis performed using apheresis equipment to separate leukocytes from peripheral blood, at the same time returns autologous plasma, platelets and erythrocytes to the patient, is applied clinically for the treatment of hyperleukocytosis. Leukapheresis not only removes excessive leukocytes rapidly and corrects metabolic abnormalities but also alleviates the symptoms of leukostasis. In addition, the procedure of leukapheresis is generally well tolerated. Leukapheresis has become one of the most imperative adjuvant therapies to treat hyperleukocytosis, especially in the patient who was not inappropriate to cytoreduce with Ara-C or hydroxyurea. In this review, we present the background of leukapheresis development and highlight its clinical application in hyperleukocytic leukemia patients.Keywords: hyperleukocytosis, leukapheresis, leukostasis, hyperleukocytic leukemia

Ikhwan Rinaldi,1 Resti Mulya Sari,2 Vanya Utami Tedhy,3 Kevin Winston3 1Division of Hematology and Medical Oncology, Department of Internal Medicine, Cipto Mangunkusumo National General Hospital, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; 2Division of Hematology and Medical Oncology, Department of Internal Medicine, Dharmais National Cancer Hospital, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; 3Faculty of Medicine, Universitas Indonesia, Jakarta, IndonesiaCorrespondence: Ikhwan RinaldiDivision of Hematology and Medical Oncology, Department of Internal Medicine, Cipto Mangunkusumo National General Hospital, Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, IndonesiaTel +628111001758Email ikhwan.rinaldi@ui.ac.idIntroduction: Leukostasis is a medical emergency with high mortality which often occurs in acute myeloid leukemia patients with hyperleukocytosis. One of the therapies that can be used for leukostasis in acute myeloid leukemia is leukapheresis. However, whether leukapheresis can provide better survival benefit when compared with patients not receiving leukapheresis is still unclear. Hence, we aimed to evaluate the effect of chemotherapy plus leukapheresis combination versus chemotherapy only on 28-day survival of acute myeloid leukemia patients with leukostasis.Methods: This study was a dual-center retrospective cohort using secondary data from medical records collected from November 2018 to March 2019. Inclusion criteria were adult patients aged 18 years old or above, diagnosed with acute leukemia with hyperleukocytosis status defined by WBC count greater than 100,000/uL, and with symptoms of leukostasis. One-month survival analysis was conducted using Kaplan–Meier curve method. Univariate and multivariate analyses were then conducted using Cox proportional hazards model to obtain value of hazard ratio (HR) with a 95% confidence interval (CI).Results: A total of 38 patients were obtained for analysis. The median overall survival was 25 days (95% CI: 17.001– 32.999 days) in the chemotherapy only group and 20 days (95% CI: 1.497– 38.503) in the chemotherapy with leukapheresis group. The use of leukapheresis did not affect 28-day survival (HR: 1.140; 95% CI: 0.396– 3.283; p value: 0.809) and 7-day survival (HR: 1.073; 95% CI: 0.277– 4.152; p value: 0.919). In the multivariate analysis, age ≥ 60 years, blast percentage ≥ 90%, creatinine ≥ 1.4 mg/dL, and presence of disseminated intravascular coagulation were associated with worse 28-day survival.Conclusion: AML patients with leukostasis who received both chemotherapy and leukapheresis did not have better 28-day survival and 7-day survival when compared with patients receiving chemotherapy only. Old age, high blast percentage, high creatinine, and presence of disseminated intravascular coagulation were prognostic factors for worse 28-day survival.Keywords: leukemia, hyperleukocytosis, leukostasis, leukapheresis, chemotherapy, survival

INTRODUCTION: Peripheral blood stem cell has become the preferred source for hematopoietic stem cells in both autologous and allogenic transplants. CD34+ cells represent a small proportion of the leukapheresis product content. The present study aims to analyze the cellular composition of leukapheresis product collected by intermittent-flow Mobile Collection System®+ (MCS+) and continuous-flow (Spectra Optia ®) apheresis equipment. METHODS: In this retrospective study, 97 leukapheresis procedures for 85 individuals mobilized only with granulocyte colony-stimulating factor were analyzed. The product samples were analyzed for CD34+ cells, red blood cells (RBCs), white blood cells, and platelet contents. The difference in overall product composition was compared between the equipment through Mann–Whitney U-test. RESULTS: Both the equipment had similar CD34 and mononuclear cells (MNC) harvested in the product. However, Spectra Optia had statistically significantly lower product volume than MCS+ (215 mL vs. 260 mL, P = 0.04). Similarly non-CD34 composition such as RBC content per apheresis was six-fold higher (27 mL vs. 4 mL) and platelet contamination (1601 vs. 1275 × 109/L) was relatively higher with MCS+ over Spectra Optia. No relationship was observed between the CD34 concentration in the product and RBC and platelet contamination between both the equipment. CONCLUSION: Both equipment collect adequate CD34 and MNC cells; however, Spectra Optia is preferred due to product quality in terms of less product volume with minimal RBC and platelet contamination.

Therapeutic leukapheresis is performed in patients having hyperleukocytosis, leading to leukostasis. The aim of this study was to evaluate the efficacy of therapeutic leukapheresis in reducing white cell count in leukemic patients with symptoms of leukostasis. Two cases of acute lymphoblastic leukemia, admitted to the hematology intensive care unit between January and May 2015, were taken up for therapeutic leukapheresis, using COBE Spectra Cell Separator. The first patient was a 24-year-old male, admitted with fever, weakness, headache, blurred vision, and a total leukocyte count (TLC) of 564,000/μL. The second patient was a 36-year-old male admitted with fever, headache, dyspnea, and a TLC of 530,000/μL. The first patient had a 31.38% and 30.19% reduction in TLC after first and second procedures, respectively. The second patient had a reduction in TLC of 31.7% and 31.41% after first and second procedures, respectively. This significant decrease in white cell count achieved reversal of symptoms of leukostasis. Patients with hyperleukocytosis may present as a medical emergency, requiring prompt recognition and initiation of therapy to prevent respiratory failure or intracranial hemorrhage. Leukapheresis removes circulating blasts quickly to alleviate symptoms.

Transfusion of granulocyte concentrates (GC) is an alternative therapy for neutropenic patients with life-threatening infections. While neutrophils are the main source of antimicrobial activity, only neutrophil numbers are used to certify GCs. The objective of this study was thus to functionally characterize neutrophils in GCs prepared by leukapheresis from G-CSF-stimulated donors and compare to the less characterized prednisone GCs. GCs prepared from healthy donors stimulated with prednisone and then G-CSF after a 6-month washout period were analyzed prior to and after leukapheresis, and after storage. Leukocyte composition, neutrophil viability, calcium mobilization, chemotaxis, phagocytosis, reactive oxygen species, cytokine production and metabolites were determined. G-CSF GCs contained significantly more neutrophils than prednisone GCs of which 40% were immature. In comparison to non-stimulated healthy donor neutrophils, prednisone GC neutrophils exhibited enhanced phagocytosis and G-CSF GC neutrophils showed decreased chemotaxis but increased IL-8 production. Leukapheresis altered prednisone GC neutrophil responses. Storage had a significant, negative impact on G-CSF GC neutrophils compared to prednisone GC neutrophils. G-CSF and prednisone GC neutrophils thus differ in maturity and function, and G-CSF GC neutrophils are more sensitive to storage. Functional testing of GC neutrophils and better storage conditions would improve the quality of this blood product.

Abstract Background Osteosarcoma (OSA) in dogs is an aggressive bone tumor with frequent chemotherapy failure and translational relevance for human health. Hypothesis/Objectives We hypothesized that dogs with OSA could be treated safely by ex vivo activated T‐cells that were generated by autologous cancer vaccination and supported by interleukin‐2 (IL‐2) treatment with survival more than twice that reported for amputation alone. Animals Osteosarcoma‐bearing dogs (n = 14) were enrolled in a single‐arm prospective trial after complete staging before amputation. Four healthy dogs also were treated in a safety study. Methods Autologous cancer cell vaccinations were administered intradermally and dogs underwent leukapheresis. Mononuclear cell products were stimulated ex vivo with a T‐cell‐activating agent. Activated product was transfused and 5 SC IL‐2 injections were administered q48h. Dogs were monitored for metastasis by thoracic radiography every 3 months. Results Autologous cancer cell vaccine and activated cellular therapy (ACT) products were successfully generated. Toxicity was minimal after premedicants were instituted before ACT. With premedication, all toxicities were grade I/II. Median disease‐free interval for all dogs was 213 days. One dog developed cutaneous metastasis but then experienced spontaneous complete remission. Median survival time for all dogs was 415 days. Five dogs survived >730 days. Conclusions and Clinical Importance This immunotherapy protocol without cytotoxic chemotherapy is safe and tolerable. Compared to historical amputation reports, survival was notably prolonged in this group of patients. Additional prospective studies are warranted to elucidate active immunologic mechanisms and further improve disease response and survival.

Aim To present characteristics of collecting autologous peripheral blood stem cells over a one-year period with an emphasis on efficiency and safety. Methods A retrograde analysis of 24 leukapheresis in 20 adult patients with malignant haematological diseases in the Blood Transfusion Institute of the Federation of Bosnia and Herzegovina in Sarajevo, was done. Cell separators Amicus and Spectra Optia were used for collection procedures. Results The patient’s age ranged from 27 to 65 years. Target cells were collected in one procedure in sixteen patients, while in four patients they were collected in two procedures. The mean CD34+ collection efficiency was 57.7%. The median number of CD34+ cells and percentage of CD34+ cells in the products were 5.52x10e6/kg (range 3.28-9.00) and 1.57% (range 0.96-2.91). A strong positive correlation was found between the number of CD34+ cells in peripheral blood on the apheresis day and the amount of CD34+ cells collected in the products (rs=0.73). A total of 95% of patients collected the amount of ≥3x10e6/kg and 55% of ≥5x10e6/kg CD34+ cells for a single transplant. A decrease in platelet count, haemoglobin and haematocrit values after the procedure was not significant. Potassium decrease showed statistical significance (p

Hyperleukocytosis in pediatric acute leukemia is associated with increased morbidity and mortality and at present there is no consensus on the use of leukapheresis (LPH) for its management. Our aim was to review characteristics and outcomes of newly diagnosed leukemia patients with hyperleukocytosis (HL) comparing those who received LPH and those who did not. An IRB approved retrospective case control study reviewed data from a single institution over a 10 year period. At our institution, LPH was used in 8 of 62 (13%) patients with hyperleukocytosis with minimal complications. Mean leukocyte count in patients who received LPH versus those who did not was 498 k cells/mm3 and 237 k cells/mm3, respectively. Patients who had symptoms of neurologic (63 vs. 17%) or pulmonary leukostasis (75 vs. 17%) were more likely to have undergone leukapheresis. The time from presentation to the initiation of chemotherapy was not different between those who received LPH and those who did not (mean of 35 h vs. 34 h). There was one death in the LPH group, that was the result of neurologic sequelae of hyperleukocytosis and not LPH itself. The use of LPH in patients with hyperleukocytosis is safe, well tolerated and does not alter time to chemotherapy at our institution.

Incidenca levkemij, odkritih v nosečnosti, znaša od 1/10.000 do 1/100.000. Nemalokrat je ob odkritju bolezni z zdravljenjem s citostatiki potrebno začeti takoj. Citostatiki so za plod teratogeni, zato moramo pred pričetkom zdravljenja pretehtati vse možnosti in izbrati najbolj sprejemljivo. Predstavljamo primera akutne mieloične levkemije (AML) in kronične mieloične levkemije (KML), odkrita v nosečnosti, ki smo ju obravnavali v Univerzitetnem kliničnem centru v Ljubljani. Gre za edina primera v zadnjih desetih letih v Sloveniji. Bolnico z AML, podvrsta akutna promielocitna levkemija (APL), smo ob začetku 2. trimesečja nosečnosti zdravili z daunorubicinom in all-transretinoično kislino (ATRA). Glede na našo izkušnjo se zdi, da so priporočila za zdravljenje APL v nosečnosti realna, izvedljiva in predvsem varna za bolnico in plod. Pri bolnici s KML, ki je prvič prišla k nam tik pred porodom, je bila v ospredju izrazita levkocitoza 335 × 109/L, trombocitoza in splenomegalija. Zavračala je vse načine zdravljenja, v prvi vrsti levkoferezo. Kljub vsemu je brez porodnih zapletov rodila zdravega otroka. Porod je bil zaradi povečane vranice in posledične nevarnosti za njeno rupturo dokončan s carskim rezom. Na osnovi naše izkušnje lahko zaključimo, da ni moč priporočiti, pri kako velikem številu levkocitov je za mati in plod koristno začeti zdraviti z levkoferezo. Enako tudi ni jasno, kakšno tveganje za rupturo vranice v teh primerih predstavlja poskus vaginalnega poroda. Uganka ostaja tudi vprašanje, kolikokrat se zaradi levkemij različnih oblik poveča tveganje za trombembolični dogodek v nosečnosti, ki že sama po sebi nagiba k trobozam.

Background: The treatment of relapsed or refractory leukemia remains a major problem. Among the new therapeutic approaches, the use of modified T lymphocytes, called chimeric antigen receptor T cells (CAR-T cells), seems promising. The first step of their preparation is leukapheresis, which involves the collection of mononuclear cells from the patient. This medical procedure requires numerous medical devices (MDs) made of plasticized polyvinylchloride (PVC). These compounds can leach out of the devices during contact with the patient’s blood. The aim of our study was to evaluate the migration of the plasticizers contained in the MD during a simulated pre-CAR-T cell leukapheresis procedure, and to measure the patient’s and their lymphocytes’ exposure to them. Methods: The qualitative and quantitative composition of the MD used for pre-CAR-T cell apheresis was determined by gas chromatography–mass spectrometry (GC–MS). Then, an ex vivo leukapheresis model using an ethanol/water simulant was performed to evaluate the plasticizers’ migration under simulated clinical conditions of pre-CAR-T cells’ cytapheresis. The plasticizers released into the simulant were quantified by GC–MS. Results: Diethylhexylphthalate (DEHP) was found in the apheresis kit, with amounts ranging from 25% to 59% (g/100 g of PVC). Bis(2-ethylhexyl) adipate was detected at trace levels. A total of 98.90 ± 11.42 mg of DEHP was released into the simulant, corresponding to an exposure dose of 1.4 mg/kg for a 70 kg patient. Conclusions: Patients undergoing a pre-CAR-T cell apheresis are mainly exposed to DEHP, which can impact their health because of its endocrine disruption effect, but could also lead to a decrease in CAR-T cells’ efficiency/quality.

Leukapheresis has been used widely to quickly reduce white blood cell count (WBC) in patients with hyperleukocytosis. Despite its wide utilization, leukapheresis has risks of complication, which have not been thoroughly reported in pediatric patients. No report was found in English literatures about leukapheresis complications in children with acute lymphoblastic leukemia (ALL).We reported a 4-year-old girl with ALL and hyperleukocytosis, with no sign of bleeding. After leukapheresis, WBC and platelet count decreased, and patient complained of headache. Neurological deficit developed quickly afterwards. Intracranial infection was suspected clinically. Contrast-enhanced head CT revealed multiple hyperdense lesions and diffuse cerebral oedema. The patient was subsequently diagnosed with multiple intraparenchymal hemorrhage, and leukapheresis was stopped.Intracranial hyperdense lesion in leukemic patients has many differential diagnosis, such as infection, granulocytic sarcoma (chloroma), and hemorrhage. Lesion characteristics in CT could help point the diagnosis. History of leukapheresis should also raise suspicion of hemorrhage. This article discussed CT characteristics of multiple brain hemorrhage in pediatric ALL after leukapheresis and how to differentiate it with other common intracranial complications of leukemia.

Aim. To identify a parameter predicting a collection of at least 2·106 CD34+ hematopoietic stem cells (HSC)/kg body weight per leukapheresis (LA) procedure. Subjects and methods. The investigation included 189 patients with hematological malignancies and 3 HSC donors, who underwent mobilization of stem cells with their subsequent collection by LA. Absolute numbers of peripheral blood leukocytes and CD34+ cells before a LA procedure, as well as a number of CD34+ cells/kg body weight (BW) in the LA product stored on the same day were determined in each patient (donor). Results. There was no correlation between the number of leukocytes and that of stored CD34+ cells/kg BW. There was a close correlation between the count of peripheral blood CD34+ cells prior to LA and that of collected CD34+ cells calculated with reference to kg BW. Conclusion. The optimal absolute blood CD34+ cell count was estimated to 20 per µl, at which a LA procedure makes it possible to collect 2·106 or more CD34+ cells/kg BW.

Extracorporeal photochemotherapy (ECP) is considered as an immune modulating therapy majorly targeting the T cells of the Immune system. ECP induces an anti-inflammatory condition with tolerogenic responses without inducing a global immunosuppression state which is a typical feature of other therapeutic options such as steroids. Clinical indication of ECP has grown over time since its initial applications. Our review discusses the technical aspects of the concept of photopheresis with the available methods for its clinical applications.

Patients presenting with hyperleukocytosis secondary to acute leukemia, with total leukocyte count or blast count more than 100,000/μL are often considered for leukapheresis, especially if clinical signs of leukostasis are present. Leukostasis is often associated with high morbidity and mortality in patients with leukemic processes. The main goal of management of hyperleukocytosis and/or leukostasis is to reduce the blast count before initiation of chemotherapy. Leukapheresis is often used prophylactically to prevent leukostasis or to provide symptomatic relief. We, as transfusion medicine specialists, present our experience of doing therapeutic leukapheresis in patients presenting with hyperleukocytosis with or without presenting features of leukostasis.

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 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.

Background: Harvest of hematopoietic progenitor cells via leukapheresis is being used increasingly for transplants in India. Adequate yield of cells per kilogram body weight of recipient is required for successful engraftment. Collection efficiency (CE) is an objective quality parameter used to assess the quality of leukapheresis program. In this study, we calculated the CE of the ComTec cell separator (Fresenius Kabi, Germany) using two different formulae (CE1 and CE2) and analyzed various patient and procedural factors, which may affect it. Materials and Methods: One hundred and one consecutive procedures in 77 autologous donors carried out over 3 years period were retrospectively reviewed. Various characteristics like gender, age, weight, disease status, hematocrit, preprocedure total leukocyte count, preprocedure CD34 positive (CD34+) cells count, preprocedure absolute CD34+ cell count and processed apheresis volume effect on CE were compared. CE for each procedure was calculated using two different formulae, and results were compared using statistical correlation and regression analysis. Results: The mean CE1 and CE2 was 41.2 and 49.1, respectively. CE2 appeared to be more accurate indicator of overall CE as it considered the impact of continued mobilization of stem cells during apheresis procedure, itself. Of all the factors affecting CE, preprocedure absolute CD34+ was the only independent factor affecting CE. Conclusion: The only factor affecting CE was preprocedure absolute CD34+ cells. Though the mean CE2 was higher than CE1, it was not statistically significant.

Introduction: 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

Patients with acute myeloid leukemia (AML) and hyperleukocytosis, defined as an initial white blood cell (WBC) count of ≥ 100 × 109/L, are often treated with leukapheresis. In this study, we have reported our experience of treating AML without leukapheresis. Methods: From November 1, 1995, to May 31, 2012, there were 74 children (≤18 years old) with de novo AML other than acute promyelocytic leukemia. Seventeen patients had an initial WBC count ≥ 100 × 109/L. Prompt chemotherapy was started within hours whereas leukapheresis was not performed. Results: The median age of the 17 patients with hyperleukocytosis was 7.4 years (range: 0–16 years), and the median initial WBC count was 177 × 109/L (range: 117–635 × 109/L). The median time between admission and initiation of chemotherapy was 4.5 hours (range: 2–72 hours) in patients with hyperleukocytosis, whereas it was 13 hours (range: 2–120 hours) in those without hyperleukocytosis. Seven patients (7/17, 41%) had one or more early complications before or during the first 2 weeks of chemotherapy. Fifteen of the 16 patients who received prompt chemotherapy achieved complete remission (93.8%), comparable with those without hyperleukocytosis (98.2%; p = 0.33). Conclusion: Children with AML and hyperleukocytosis, treated with prompt chemotherapy without leukapheresis, had minimal early morbidities.

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 × 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: Pre-mobilization 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 independent factor 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.

Objective: To evaluate factors affecting peripheral bloodhematopoietic stem cell yield in patients undergoing large-volumeleukapheresis for autologous peripheral blood stem cell collection.Methods: Data from 304 consecutive autologous peripheral bloodstem 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 106CD34+ cells/kg of body weight. Pre-mobilization factors analyzedincluded patient’s age, gender and diagnosis. Post mobilizationparameters evaluated were pre-apheresis peripheral white bloodcell count, immature circulating cell count, mononuclear cell count,peripheral blood CD34+ cell count, platelet count, and hemoglobinlevel. 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 independent factor 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.

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 ≥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≤4) in 70.2% patients (33/47). The number of submissions for colectomy was higher for severe UC at entry (CAI≥12, n=25) than for moderately severe UC at entry (7≤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. (Gut and Liver 2009;3:41-47)

Age-associated decline of immune function is believed to be largely due to alterations of immune cell regeneration. However, longitudinal changes of human immune cells with age have not yet been adequately addressed. To test the hypothesis that regeneration of lymphocytes and monocytes is robust through most of adult life until advanced age, we examined six leukapheresis donors (3 young and 3 middle-aged/old) who donated approximately 10% of their peripheral blood mononuclear cells (PBMC) every other month over 3-5 years. We found the number of both lymphocytes and monocytes were quite stable in the blood of all six donors. As expected, young donors had more TRECs, more CD31+ cells (CD4 only) and longer telomeres in T cells than did the middle-aged donors. Interestingly, more variation in TREC number, Vβ usages, and telomere lengths were observed in young donors during the 3-5 years course of donation whereas the middle-aged/old donors showed a rather striking stability in these measurements. This may reflect a more prominent role of thymic output in T cell regeneration in young than in middle-aged/old donors. Together, these findings provide an in vivo glimpse into the homeostasis of lymphocytes and monocytes in the blood at different ages, and support the notion that regeneration of lymphocytes and monocytes is robust throughout adult life up to the early 70s.

Circulating tumor cells (CTCs) are believed to be responsible for the development of metastatic disease. Over the last several years there has been a great interest in understanding the biology of CTCs to understand metastasis, as well as for the development of companion diagnostics to predict patient response to anti-cancer targeted therapies. Understanding CTC biology requires innovative technologies for the isolation of these rare cells. Here we review several methods for the detection, capture, and analysis of CTCs and also provide insight on improvements for CTC capture amenable to cellular therapy applications.

Background. In the study we tried to define a predictive value of the circulating CD34+ cells in patients/ donors blood for estimation of the hematopoietic stem cells (HSC) collection efficacy determine the optimal time to initiate the collection by leukapheresis procedure.Methods. We retrospectively analyzed 75 collections of HSC using the Amicus cell separator in 39 patients and 15 donors. Circulating CD34+cell counts in patients/donors were compared to the achieved CD34+ cell yields to determine its predictive value for the collection of a targeted yield of > 2 × 106 CD34+ cells/kg body weight of patient.Results. The results of cell counts confirmed that mobilization regimens were successful and HSC collections efficient. High correlation coefficient (r = 0.82) between the number of circulating CD34+ cells before collection and CD34+ cell yield/kg of patient’s body weight was statistically significant (p < 0.05). With ROC analysis we determined the cut-off value 42 × 106/l CD34+ cell counts in the blood of patients/donors before collection that had a positive predictive value 87% and a negative predictive value 91.6%.Conclusions. Analysis showed that the number of circulating CD34+ cells before the procedure express a very high predictive value and can be used for determining the optimal time to initiate collection of HSC by leukapheresis.

Cell separators in India are routinely used for plateletpheresis, peripheral blood stem cell collections and therapeutic plasma exchange. Therapeutic leukapheresis, particularly as an emergency procedure, has been uncommonly performed and reported. Here, a case of a 53-year-old male, diagnosed with acute myeloid leukemia subtype M5 (AML M5) with hyperleukocytosis, who underwent emergency leukaphereis, is reported. After two procedures, there was a decrease of WBC count by 85%, which enabled cytotoxic therapy to be initiated.

Se realizó un estudio descriptivo de la totalidad de las colectas de células progenitoras hematopoyéticas realizadas en el Banco de Sangre del Hospital "Hermanos Ameijeiras" desde abril del 2001 hasta abril del 2002, mediante leucoféresis. Se utilizó un equipo de flujo continuo (FRESENIUS ASTEC 204) y se empleó el programa mononucleares con el set P1Y. Se determinó peso, talla, hematócrito, hemoglobina, conteo de leucocitos y plaquetas de los donantes previo a la colecta. En el producto de la leucoféresis se analizó el conteo de leucocitos con diferencial, conteo de células CD34+/ CD45+ (mediante técnica de inmunofluorescencia en citómetro de flujo) y determinación de viabilidad celular con yoduro de propidio en este mismo equipo (Facscan, Becton Dickinson). Recibieron trasplante autólogo 5 pacientes y 2 trasplantes alogénicos. Los primeros con diagnósticos de: linfoma no hodgkiniano (3), enfermedad de Hodgkin (1), artritis reumatoidea (1); y los segundos afectados de leucemia mieloide crónica (1) y linfoma no hodgkiniano (1) ; todos ellos atendidos en el Servicio de Hematología de nuestro centro. La media del conteo de células CD34+ por cada leucoféresis para trasplante autólogo fue 2,4 x 106 Kg y en el alogénico 2,96 x 106 Kg . Los resultados evidencian que mediante el ajuste del volumen de los ciclos, la velocidad centrífuga, el volumen de rebosado y el volumen celular, se obtuvo un conteo de células precursoras hematopoyéticas en la colecta similar a lo reportado por otros autores, lo que garantiza que el producto transfundido cuente con la celularidad necesaria para repoblar la médula ósea, luego que el paciente sea tratado con altas dosis de terapia inmunosupresora y/o radiacionesA descriptive study of all the collections of hematopoietic progenitor cells obtained by leukophoresis at the Blood Bank of "Hermanos Ameijeiras" Hospital from April, 2001, to April, 2002, was conducted. A continual flow equipment (FRESENIUS ASTEC 204) was used and the mononuclear program with the P1Y set was applied. Weight, height, haematocrit, haemoglobin, leukocyte and platelet count of the donors were determined before the collection. In the product of the leukophoresis, the leukocyte count with differential, the CD34+/CD45+ cells count (by immunofluorescence technique in flow cytometer) and the determination of cellular viability with propidium iodide in this equipment (Facscan, Becton Dickinson), were analyzed. 5 patients recieved autologous transplantations and 2 allogeneic transplantations. The first were diagnosed non-Hodgkin lymphoma (3), Hodgkin disease (1), and rheumatoid arthritis (1); whereas the others were affected by chronic myeloid leukemia (1) and non-Hodgkin lymphoma. All of them were attended at the Hematology Service of our center. The mean of the CD34+ cell count per each leukophoresis for autologous transplantation was 2.4 x 106 Kg, whereas in the allogeneic it was 21.96 x 106 Kg. The results show that by the adjustment of the cycle volume, the centrifugal speed, the overflowing volume and the cellular volume, it was possible to obtain a count of haematopoietic precursor cells in the collection similar to the reported by other authors, guaranteeing that the transfused product has the necessary cellularity to repopulate the bone marrow after the patient is treated with high doses of immunosuppressor therapy and/or radiations