No abstract available.
Neoplasm, Residual*

BACKGROUND: Peripheral blood progenitor cells collected from normal donors after granulocyte- colony stimulating factor (G-CSF) treatment are increasingly used for allogeneic transplantation. Previously, we investigated the mobilization kinetics of CD34+/Thy-1dim progenitor cells during subcutaneous administration of G-CSF in normal donors (Transfusion 1997;37:406-10). Although it is considered to be a relatively safe procedure, there are still uncertainties about the most efficient method of progenitor cell mobilization. Due to the short elimination half-life of G-CSF of about 3-4 hours we considered the subcutaneous administration of G-CSF once or twice daily might be suboptimal. The aim of the present study is to evaluate the mobilization kinetics of CD34+ cells during continuous intravenous (IV) administration of G-CSF in normal donors. METHODS: Fifteen healthy donors were enrolled in this study. The median age was 38 years (range, 20-56). G-CSF (Filgrastim, 10 microgram/kg/day) was administered for 4 consecutive days through continuous IV infusion. Then, we collected PBPC on the day following the 4th dose of G-CSF using a blood cell separator. For measurement of complete blood counts and CD34+ cell levels, peripheral blood sampling was performed immediately before the administration of G-CSF (steady-state) and after 4, 8, 24, 48, 72, 96, 120 hours of continuous IV administration of G-CSF. RESLUTS: After continuous IV administration of G-CSF, the WBC counts increased up to day 5 and reached approximately 8.4-fold above the steady-state level. Changes in the granulocyte count were similar to those in WBC counts. The number of lymphocytes increased up to day 4 (2.7-fold above the steady-state level), but no further increase occurred on day 5. Although there were considerable variations among the healthy donors, the statistical peaks of CD34+ cell levels were consistently observed on day 3 or day 4. Up to the fourth day of G-CSF treatment, the circulating CD34+ cells expanded by 25-fold. The percentage and absolute number of CD34+ cells significantly increased on day 3 (0.55 +/- 0.09%, 51.12 +/- 24.83x103/mL) and day 4 (0.47 +/- 0.09%, 46.66 +/- 24.93x103/mL), compared with steady-state values (0.06 +/- 0.09%, 2.03 +/- 5.69x103/mL). CONCLUSION: Our results showed that continuous IV administration of G-CSF apparently results in more rapid mobilization of CD34+ cells at least 24-48 hours compared with daily subcutaneous administration of G-CSF in normal donors.
Administration, Intravenous* ; Blood Cell Count ; Blood Cells ; Colony-Stimulating Factors ; Granulocyte Colony-Stimulating Factor* ; Granulocytes ; Half-Life ; Humans ; Kinetics* ; Lymphocytes ; Stem Cells ; Tissue Donors* ; Transplantation, Homologous

No abstract available.
Stem Cells*

BACKGROUND: The stromal cells, which one part of the microenvironment, including adipocyte, fibroblasts, endothelial cells, macrophage and smooth muscle cells provide a suitable environment for self-renewal, proliferation and differentiation of hematopoietic stem cells, as well as providing other regulators, cytokines to stimulate and enhance hematopoiesis. Unlike bone marrow, the in vitro development of this stroma cells from the umbilical cord blood (UCB) is not well established and has met very limited success. To see if stromal elements capable of supporting hematopoiesis could be grown from CD34+ enriched umbilical cord blood, we was done stroma-free liquid culture system with thrombopoietin (TPO), flt3-ligand (FL) and the granulocyte- colony stimulating factor (G-CSF) using cord blood CB CD34+ cells. METHODS: Purified umbilical cord blood CD34+ cells were cultured for five weeks in a stroma-free liquid culture system using TPO, FL, and G-CSF in the density of 1x105 cells/mL. When a firmly adherent monolayer of fibroblast like cells were fully formed, we evaluate morphology, immunopheno-type, immunohistochemistry and functional assay, including non-adherent expansion, clonogenic progenitor expansion and apoptosis of established adherent cell layers. RESULTS: By the 4th~5th week, we found that firmly adherent fibroblast-like cells were established. These cells showed characteristics of endothelial cells expressing with VWF, VCAM-1, ICAM-1, CD31 and E-selectin. Furthermore, comparing without established endothelial monolayer, with established endothelial monolayer better expansion of total nucleated cells, CD34+ cells and colony forming units granulocyte-macrophage (CFU-GM) during culture were found. CONCLUSION: These results suggest that the UCB CD34+ cell fraction contains stromal cell precursors and can establish the hematopoietic microenvironment to expand and sustain UCB CD34+ cells through down-regulating apoptosis.
Adipocytes ; Apoptosis ; Bone Marrow ; Colony-Stimulating Factors ; Cytokines ; E-Selectin ; Endothelial Cells ; Fetal Blood* ; Fibroblasts ; Granulocyte Colony-Stimulating Factor* ; Hematopoiesis ; Hematopoietic Stem Cells ; Humans* ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; Macrophages ; Myocytes, Smooth Muscle ; Stem Cells ; Stromal Cells ; Thrombopoietin ; Vascular Cell Adhesion Molecule-1

BACKGROUND: The aim of the present study is to evaluate the modulation of CD44 and CD31 expression and apoptotic status on mobilized CD34+ cells during continuous intravenous (i.v.) administration of granulocyte-colony stimulating factor (G-CSF), elucidating the mechanism of peripheral blood progenitor cells (PBPC) mobilization in normal donors. METHODS: Fifteen healthy donors were enrolled in this study. G-CSF (10 microgram/kg/day) was administered for 4 consecutive days through continuous i.v. infusion. Measurement of CD34+ cell levels and their expression of CD44, CD31 and 7-aminoactinomycin D (7-AAD) was performed. PB sampling was drawn immediately before the administration of G-CSF (steady-state) and after 4, 8, 24, 48, 72, 96 and 120 hours of G- CSF administration. RESULTS: Although there were considerable variations among the healthy donors, the absolute number of CD34+ cells significantly increased at day 3 (51.12+/-24.83x103/mL) and day 4 (46.66+/-24.93x103/mL),compared to the steady-state level (2.03+/-5.69x103/mL). The expression of CD44 on CD34+ cells revealed a significant reduction from the steady-state level (579.59+/-133.69) after day 3 (281.02+/-105.15, P=0.0059) and day 4 (164.76+/-107.44, P=0.0002) of G-CSF administration. The expression of CD31 on CD34+ cells also significantly decreased from the steady-state level (145.86+/-30.52) after day 4 (34.47+/-38.87, P=0.0055) and day 5 (17.33+/-50.68, P=0.0134) of G-CSF administration. The proportions of apoptotic (7-AADdIm) CD34+ cells were also significantly decreased after day 3 of G-CSF administration. CONCLUSION: Down-regulation of CD44 and CD31 on CD34+ cells is likely to be involved in the mobilization of PBPC and G- CSF may acts as a survival factor for mobilized CD34+ cells by the suppression of apoptosis during continuous i.v. administration of G-CSF in normal donors.
Administration, Intravenous* ; Apoptosis ; Down-Regulation ; Granulocyte Colony-Stimulating Factor ; Humans ; Stem Cells ; Tissue Donors*

Emergence of new clonal chromosomal abnormality (CCA) has been reported in Philadelphia-negative cells in patients with chronic myeloid leukemia (CML) undergoing the tyrosine kinase inhibitor (TKI) treatment. However, the time of emergence and clinical significance of CCA remains to be elucidated. In this study, we report a CML patient undergoing TKI treatment who developed myelodysplastic syndrome (MDS) after 206 months since the diagnosis of CML. Results of droplet digital PCR performed with serial bone marrow samples revealed that monosomy 7 in Philadelphia-negative cells appeared at the time of MDS development that did not exist initially at the time of CML diagnosis.

Emergence of new clonal chromosomal abnormality (CCA) has been reported in Philadelphia-negative cells in patients with chronic myeloid leukemia (CML) undergoing the tyrosine kinase inhibitor (TKI) treatment. However, the time of emergence and clinical significance of CCA remains to be elucidated. In this study, we report a CML patient undergoing TKI treatment who developed myelodysplastic syndrome (MDS) after 206 months since the diagnosis of CML. Results of droplet digital PCR performed with serial bone marrow samples revealed that monosomy 7 in Philadelphia-negative cells appeared at the time of MDS development that did not exist initially at the time of CML diagnosis.

Cases of clonal cytogenetic abnormalities in Philadelphia-negative cells during the treatment of Philadelphia-positive CML have been previously reported. However, clonal abnormalities unrelated to the original t(8;21) or t(15;17) karyotype are not common. Deletion of 20q (del(20q)) is one of the most common recurrent cytogenetic abnormalities in myeloid neoplasms. Here we describe 3 patients with t(8;21), t(15;17), or t(9;22) who developed unrelated del(20q) after successful treatment of leukemia. We retrospectively reviewed the cytogenetic results of 23 AML patients with t(8;21)(q22;q22), 28 AML patients with t(15;17)(q22;q12), and 47 CML patients with t(9;22)(q34;q11.2). We identified 3 patients with del(20q) as the only clonal aberration unrelated to the primary karyotype when they achieved complete morphologic and cytogenetic remission. The latency period between diagnosis and emergence of del(20q) was 1, 114, and 35 months for the 3 patients, respectively. There was no evidence of therapy-related MDS/AML during the follow-up period. In 1 AML patient with t(8;21), relapse occurred in a t(8;21)(q22;q22) clone and the del(20q) clones were lost. The clinical significance of del(20q) as an unrelated clonal aberration is unknown, but our study suggests that del(20q) does not cause therapy-related MDS/AML or indicate disease progression.
Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Human, Pair 20 ; Clone Cells ; Cytogenetics ; Disease Progression ; Follow-Up Studies ; Humans ; Karyotype ; Latency Period (Psychology) ; Leukemia ; Recurrence ; Retrospective Studies

BACKGROUND: Recently, the commercial kits for measurement of the absolute number of CD34+ cells have been introduced as a standard method. The aims of this study was to investigated optimal timing of peripheral blood progenitor cell(PBPC) collection and optimal CD34+ cells dose transplanted by measurement of the absolute CD34+ cells. METHODS: We measured total leukocyte count, mononuclear cell count and the absolute number of CD34+ cells using ProCOUNT(Becton Dickinson, USA) in peripheral blood from 54 patients and 7 normal donors who underwent 101 leukapheresis for PBPC collection. We studied correlations among the absolute number of circulating CD34+ cells, other predictors and harvesting yields. We investigated relationships between the posttransplant hematopoietic recovery and CD34+ cells dose in 30 patients. RESULTS: The total number of CD34+ cells in harvesting products could be mostly predicted from the absolute number of circulating CD34+ cells. From 4 to 6 day after G-CSF mobilization, the absolute number of circulating CD34+ cells was peaked. A number of circulating CD34+ cells more than 20/microliter ensured 2.5x106 CD34+ cells/Kg in harvesting products. The patients received CD34+ cells dose >3.5x106/Kg led to a significantly faster recovery of platelets, compared with the patients receiving <3.5x106 CD34+ cells/Kg(P<0.05). CONCLUSIONS: These results suggest that PBPC collection should be started at day of circulating CD34+ cells more than 20/microliter or 4-6 days after G-CSF mobilization for successful leukapheresis and the CD34+ cell dose more than 3.5x106/Kg for PBPC transplantation could predicted rapid hematopoietic recovery.
Cell Count ; Granulocyte Colony-Stimulating Factor ; Humans ; Leukapheresis ; Leukocyte Count ; Stem Cells* ; Tissue Donors

Although neutrophilia can manifest from various causes, it is important to be able to distinguish chronic neutrophilic leukemia (CNL) from neutrophilic leukemoid reactions (NLR). In this paper, we describe four cases of leukocytosis with neutrophilia, including one case of CNL with a T618I mutation in colony stimulating factor 3 receptor (CSF3R) and three cases of NLR associated with malignancy or sepsis, which were initially suspected as CNL. Of the three NLR cases, one was associated with ovarian cancer, one with monoclonal gammopathy of undetermined significance and one with multiple myeloma with sepsis. This study demonstrated that confirming the clonality of myeloid cells with CSF3R T618I could contribute to making an accurate differential diagnosis between CNL and NLR in patients with solid cancers or plasma cell neoplasms caused by paraneoplastic syndromes and/or infection.
Colony-Stimulating Factors ; Diagnosis, Differential ; Humans ; Leukemia, Neutrophilic, Chronic* ; Leukemoid Reaction* ; Leukocytosis ; Monoclonal Gammopathy of Undetermined Significance ; Multiple Myeloma ; Myeloid Cells ; Neoplasms, Plasma Cell ; Neutrophils* ; Ovarian Neoplasms ; Paraneoplastic Syndromes ; Sepsis