Impaired Clonogenic Capacity Contributes to Bone Marrow Hypoplasia and Late Hematological Recovery after Zevalin in the Low-Grade NHL Patients.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4610-4610
Author(s):  
Wojciech Jurczak ◽  
Marta Szostek ◽  
Zbigniew Rudzki ◽  
Dorota Krochmalczyk ◽  
Joanna Wegrzyn ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Morphological Changes ◽  
Petri Dish ◽  
Relative Increase ◽  
Significant Inhibition ◽  
Low Grade ◽  
Bone Marrow Hypoplasia

Abstract Chemoimmunotherapy with Zevalin (anty CD20,90Y) is a new therapeutic approach in the resistant or relapsed low grade malignant lymphoma patients. The most important side effect observed in the up to date studies is related to transient peripheral blood cytopenias. In an attempt to evaluate the involved pathogenetic mechanisms of the above findings, the blood and bone marrow samples from 8 low grade NHL patients were prospectively tested for the peripheral blood count, the clonogenic capacity and the histopathology abnormalities (trephine biopsies) before and every 14 days after Zevalin administration until recovery. In the majority evaluated patients the mild thrombocytopenia (30-55000/μl) was observed 4 weeks after Zevalin therapy. Bone marrow mononuclear cells were cultured in methylcellulose assay using combination of recombinant CSF (IL-3, IL-6, stem cell factor, erythropoietin; Methocult, StemCell, Canada), at a concentration of 1x10^5 cells, in 35 mm gridded Petri dish and incubated at 37°C, in a humidified atmosphere of 5% CO2. The hematopoietic colonies (CFU-GM, BFU-E and CFU-GEMM) were evaluated on 14th d. of the culture. The in vitro CFU-Meg formation in the serum-free collagen-based semisolid culture (Megacult, StemCell, Canada) were also studied, stained after 12 days with a monoclonal antibody against CD 41. In the results of our studies the significant inhibition of colony formation was found already in 7th day for CFU-GM and BFU-E and starting from 14th day after Zevalin administration for CFU-Meg. In the subsequent cultures evaluated during 4–8 weeks after Zevalin therapy the three lineages hematopoietic clonogenic capacity returned to the pre-radiotherapy values. In the repeated trephine biopsis several morphological changes were observed: oedema of stromal cells, the relative increase of erythropoietic components with inverted proportion of granulo- to erythropoiesis. In conclusion, chemoimmunotherapy with Zevalin resulted in the reversible peripheral cytopenia parallely with transient damage to bone marrow hematopoiesis and in vitro clonogenic capacity.

Abstract 697: Mobilization of Oct-4 + Bone Marrow-Derived Very Small Embryonic-Like Stem Cells in Patients with Acute Myocardial Infarction

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Wojciech Wojakowski ◽  
Magda Kucia ◽  
Boguslaw Machalinski ◽  
Edyta Paczkowska ◽  
Joanna Ciosek ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Acute Myocardial Infarction ◽  
Stem Cells ◽  
Bone Marrow ◽  
Pluripotent Stem Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Small Cells ◽  
Acute Mi

Bone marrow-derived CD34 + CXCR4 + progenitor cells are mobilized into peripheral blood early in acute myocardial infarction (MI). Adult murine bone marrow contains population of small CD34 + lin − CD45 − CXCR4 + cells expressing markers of pluripotent stem cells (PSC) SSEA, Oct-4 and Nanog. This population of very small embryonic-like cells (VSEL) has unique morphology (small size 2– 4 μm, large nucleus, euchromatin) and capability to form embrioid bodies (EB). Murine EB-derived cells can in vitro differentiate into cells from all three germ layers including cardiomyocytes. We hypothesized that in patients with acute MI small cells expressing the VSEL immunophenotype and PSC markers are present in bone marrow and mobilized into peripheral blood. Blood samples (20 mL) from 18 patients with acute MI were obtained after 12 hours, 2 and 5 days after symptoms onset. Bone marrow samples (20 mL) were obtained from 2 patients with acute MI and 3 healthy volunteers. Mononuclear cells were isolated using hypotonic lysis and samples were analyzed by FACS. Mobilization of following cell populations was confirmed: hematopoietic lin − CD45 + CXCR4 + , lin − CD45 + CD133 + , lin − CD45 + CD34 + and non-hematopoietic (VSEL) lin − CD45 − CXCR4 + , lin − CD45 − CD133 + , lin − CD45 − CD34 + . Analysis of the cell number using lymphocyte gate showed more significant increase of CD45 + (hematopoietic) populations of lin − CD34 + , lin − CD133 + and lin − CXCR4 + cells. After gating for small events (VSEL size range) we found more significant mobilization of small, non-hematopoietic populations of lin − CD34 + , lin − CD133 + and lin − CXCR4 + cells (Table ). The expression of PSC markers (Oct-4, Nanog, SSEA-1) in VSEL was confirmed using real-time RT-PCR. Conclusion: We report for the first time that acute MI is associated with mobilization of non-hematopoietic VSELs expressing pluripotent stem cells markers.

SD-208, a Novel Transforming Growth Factor Beta Receptor I Kinase Inhibitor, Can Stimulate Hematopoiesis in Myelodysplastic Syndrome Progenitors.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3448-3448
Author(s):  
Amit Verma ◽  
Tony A. Navas ◽  
Jing Ying ◽  
Aaron N. Nguyen ◽  
Perry Pahanish ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Mononuclear Cells ◽  
Transforming Growth Factor ◽  
Kinase Inhibitor ◽  
Specific Inhibitor ◽  
Low Grade ◽  
Bone Marrow Biopsies

Abstract Transforming Growth Factor β (TGF-β) is a myelosuppressive cytokine that has been implicated in the ineffective hematopoiesis seen in myelodysplastic syndromes (MDS). Overactivation of TGF-β signaling in this disease was demonstrated immunohistochemically by significantly higher nuclear SMAD2 phosphorylation observed in 20 MDS bone marrows when compared with 7 non MDS anemic controls (P < 0.0001, 2 Tailed T Test, Image Pro Plus software). This data along with high levels of membrane-bound and plasma TGF-β observed in MDS patients in previous studies support the development of therapeutics targeting the TGF-β signaling pathways in this disease. SD-208 is a novel, potent and specific inhibitor of TGF-β Receptor I (TGFβ-RI) kinase. We demonstrate that SD-208 blocks the phosphorylation of SMAD2 in hematopoietic progenitors which are at the colony forming unit-erythroid (CFU-E) stage of differentiation. SD-208 also abrogates the G0/G1 cell cycle arrest induced by TGF-β in bone marrow progenitors. SD-208 treatment leads to reversal of the myelosuppressive effects of TGF-β on erythroid and myeloid colony formation from primary human CD34+ cells. Selectivity of SD-208 in inhibiting TGF-β-mediated effects on hematopoiesis was supported by similar results observed with siRNAs targeting SMAD2, a major component of the TGF-b signaling pathway. Finally, the efficacy of SD-208 in MDS was evaluated by treating bone marrow mononuclear cells from 15 patients with early low grade MDS. SD-208 treatment led to dose-dependent increases in erythroid and myeloid colonies after 14 days of in vitro culture. The effect was most notable in patients with high levels of activated SMAD-2, as assessed by immunohistochemical staining of bone marrow biopsies. Stimulation of hematopoiesis in MDS-derived marrow culture by SD-208 demonstrates a novel concept and potential therapeutic role for TGFβ-RI inhibition in this disease. Supported by VISN-17 grant, Harris Methodist Foundation Grant and ASCO YIA to AV

scholarly journals Overlapping of mononuclear cells derived from bone marrow in rats' intervertebral discs: an in vitro study

2010 ◽  
Vol 40 (4) ◽  
pp. 900-906
Author(s):  
Erica Batista Fontes ◽  
Andréa Pacheco Batista Borges ◽  
Ney Luis Pippi ◽  
Maurício Rosa ◽  
Arícia Sprada ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Intervertebral Disc ◽  
Mononuclear Cells ◽  
Morphological Changes ◽  
In Vitro Study ◽  
Intervertebral Discs ◽  
Histopathological Analysis ◽  
Disc Regeneration ◽  
Intervertebral Disc Regeneration

In this study, bone marrow mononuclear cells (BM-MNC) derived from rats were used in order to promote intervertebral disc regeneration. These cells were isolated after centrifugation in a Ficoll-Paque™ PLUS density gradient and then placed in plastic dishes to proliferate during a period of 14 days. The BM-MNCs were previously labeled with the fluorescent membrane marker Chloromethyl-benzamidodialkylcarbocyanine (CM-DIL), and thereafter were implanted in rats' intervertebral discs explants as an in vitro experimental model. Daily analyses of the cells under a fluorescence microscope revealed morphological changes, which assumed a thin and elongated shape similar to cells that originally form the annulus fibroses. Histopathological analysis demonstrated the presence of mononuclear cells interspersed within collagen fibers. The presence of viable cells, in which were found morphological changes and their disposal in the same pattern of the layers that originate the annulus fibrosus, is an indicator that they engrafted and proliferated on the intervertebral disc. Therefore, morphological changes presented by these cells indicate that they presented mesenchymal stem-like cell characteristics.

scholarly journals Increase in circulating stem cells following chemotherapy in man

Blood ◽  
1976 ◽  
Vol 47 (6) ◽  
pp. 1031-1039 ◽  
Author(s):  
CM Richman ◽  
RS Weiner ◽  
RA Yankee
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Stem Cell ◽  
Cancer Patients ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Intensive Chemotherapy ◽  
Large Numbers ◽  
Intermittent Chemotherapy

Abstract The number of circulating granulocytic stem cells (CFU-C) was determined by the in vitro methylcellulose technique in cancer patients receiving intermittent chemotherapy. In 17 patients studied prior to therapy, the median CFU-C concentration per 2 X 10(5) mononuclear cells plated was six, compared to a posttreatment median of 23 in 21 patients (p less than 0.001). Large numbers of stem cells were obtained by leukopheresis and cryopreserved with a 99.5% median CFU-C recovery. Cyclical changes in the concentration of stem cells with maximum values of 20 times baseline were demonstrated in a patient studied at weekly intervals during multiple courses of treatment. It was estimated that, at peak CFU-C concentrations, a quantity of stem cells equivalent to that present in a bulk bone marrow harvest could be obtained from the peripheral blood by a 17-liter pheresis. These results suggest that it may be practical to obtain an adequate number of stem cells from the peripheral blood to study autologous stem cell infusion as a means of averting myelosuppression in patients receiving intensive chemotherapy.

scholarly journals Abnormal in vitro proliferation and differentiation of T colony-forming cells in patients with lymphadenopathy syndrome

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1063-1069 ◽  
Author(s):  
Y Lunardi-Iskandar ◽  
V Georgoulias ◽  
W Rozenbaum ◽  
D Klatzmann ◽  
MC Coll ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Lymph Node ◽  
Growth Factors ◽  
Peripheral Blood ◽  
Colony Formation ◽  
Mononuclear Cells ◽  
Bone Marrow Cells ◽  
Interleukin 2 ◽  
Proliferation And Differentiation

Abstract Patients with acquired immunodeficiency syndrome (AIDS) present impaired colony growth and in vitro differentiation capacity of peripheral blood and bone marrow T colony-forming cells (T-CFC). We show that peripheral blood, bone marrow, and lymph node T-CFC from patients with persistent lymphadenopathy syndrome (LAS), a syndrome that can precede AIDS, displayed similar abnormalities. Indeed, peripheral blood T-CFC generated a low number of colonies in seven out of 12 patients, and almost no colonies were obtained from bone marrow cells of all patients. The simultaneous study of T-CFC from peripheral blood and lymph node mononuclear cells seems to provide a reliable indicator for the risk of developing AIDS. The six patients who developed AIDS displayed extremely low numbers of peripheral blood T- CFC (13 +/- 17 colonies per 5 X 10(4) cells), and in two of them, no colonies could be obtained from lymph node T-CFC. The remaining patients who had not developed AIDS displayed a higher number of peripheral blood T-CFC (141 +/- 113 per 5 X 10(4) cells) and lymph node T-CFC, which, in addition, preserved their clonogenic capacity. In some patients, peripheral blood and lymph node, but not bone marrow, T-CFC were capable of generating colonies in the absence of added growth factors or mitogens, whereas in others, colony formation was obtained with purified interleukin 2 (IL 2) alone. Both spontaneous and IL 2- induced colony formation was abrogated by a monoclonal antibody against the IL 2 receptor. Taken together, these findings suggest that at least some T-CFC expressed IL 2 receptors. Colonies generated either in the presence or in the absence of added growth factors were composed of T4+, T6+, and T8+ cells, indicating impaired in vitro T-CFC differentiation. These findings indicate that a dramatic quantitative and qualitative impairment of the proliferation and differentiation of peripheral blood and lymph node T-CFC precedes the clinical evolution from LAS to AIDS.

scholarly journals Hepatitis A Virus Suppresses Monocyte-to-Macrophage Maturation In Vitro

2002 ◽  
Vol 76 (9) ◽  
pp. 4350-4356 ◽  
Author(s):  
Sabina Wünschmann ◽  
Britta Becker ◽  
Angelika Vallbracht
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Hepatitis A ◽  
Mononuclear Cells ◽  
Hepatitis A Virus ◽  
Phagocytic Capacity ◽  
Maturation In Vitro ◽  
Bone Marrow Cultures ◽  
Marrow Stroma

ABSTRACT To analyze the pathogenetic mechanism of hematopoietic dysregulation associated with hepatitis A virus (HAV) infections, we studied the influence of HAV on monocyte (MO)-to-macrophage (MAC) maturation in vitro. Exposure of peripheral blood-derived mononuclear cells (MNC) to HAV led to diminished adherence of MO to plastic. Furthermore, HAV inhibited the ability of peripheral blood MO to differentiate toward MAC. Freshly isolated and 14-day-old MO cultures demonstrated reduced differentiation and decreased phagocytic capacity after challenge with HAV. Viral replication in MO/MAC cultures was confirmed by titration of infectious virus. We also determined the influence of HAV on the MO/MAC population in human long-term bone marrow cultures (LTBMCs). Inoculation of bone marrow MNC with HAV suppressed the establishment of an adherent stromal layer containing a reduced number of MAC. Furthermore, increased MO numbers in the nonadherent fraction of HAV-challenged LTBMCs are indicative of the disturbance of MO adherence. These findings suggest that HAV infection leads to a disorder of the mononuclear phagocytic system which may contribute to functional abnormalities of the bone marrow stroma.

scholarly journals THE ORIGIN AND KINETICS OF MONONUCLEAR PHAGOCYTES

1968 ◽  
Vol 128 (3) ◽  
pp. 415-435 ◽  
Author(s):  
Ralph van Furth ◽  
Zanvil A. Cohn
Keyword(s):  
Bone Marrow ◽  
Steady State ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Calf Serum ◽  
Peritoneal Macrophages ◽  
Mononuclear Phagocytes ◽  
Sterile Inflammation

The origin and turnover of efferent populations of mouse mononuclear phagocytes has been described. Mononuclear phagocytes were defined as mononuclear cells which are able to adhere to glass and phagocytize. In vitro labeling studies with thymidine-3H showed that monocytes in the peripheral blood and peritoneal macrophages do not multiply and can be considered end cells in a normal, steady state situation. However, the mononuclear phagocytes of the bone marrow appear to be rapidly dividing cells. This conclusion was supported by in vivo labeling experiments. A peak of labeled mononuclear phagocytes of the bone marrow was found 24 hr after a pulse of thymidine-3H. This was followed, 24 hr later, by a peak of labeled monocytes in the peripheral blood. From these experiments it was concluded that the rapidly dividing mononuclear phagocytes of the bone marrow, called promonocytes, are the progenitor cells of the monocytes. Labeling studies after splenectomy and after X-irradiation excluded other organs as a major source of the monocytes. Peak labeling of both the blood monocyte and peritoneal macrophages occurred at the same time. A rapid entry of monocytes from the blood into the peritoneal cavity was observed, after a sterile inflammation was evoked by an injection of newborn calf serum. These data have led to the conclusion that monocytes give rise to peritoneal macrophages. No indications have been obtained that mononuclear phagocytes originate from lymphocytes. In the normal steady state the monocytes leave the circulation by a random process, with a half-time of 22 hr. The average blood transit time of the monocytes has been calculated to be 32 hr. The turnover rate of peritoneal macrophages was low and estimated at about 0.1% per hour. On the basis of these studies the life history of mouse mononuclear phagocytes was formulated to be: promonocytes in the bone marrow, → monocytes in the peripheral blood, → macrophages in the tissue.

scholarly journals Characterization of the potentiation effect of activin on human erythroid colony formation in vitro

Blood ◽  
1989 ◽  
Vol 73 (4) ◽  
pp. 952-960 ◽  
Author(s):  
J Yu ◽  
L Shao ◽  
J Vaughan ◽  
W Vale ◽  
AL Yu
Keyword(s):  
Bone Marrow ◽  
T Lymphocytes ◽  
Dna Synthesis ◽  
Peripheral Blood ◽  
Colony Formation ◽  
Mononuclear Cells ◽  
Erythroid Progenitors ◽  
Potentiation Effect ◽  
Proliferation And Differentiation

Activin, also named FSH-releasing protein, was previously shown to induce hemoglobin accumulation in K562 cells and potentiate the proliferation and differentiation of CFU-E in human bone marrow cultures. Present studies indicate that the potentiation effect of activin is lineage specific. In addition to CFU-E, activin caused an increase in the colony formation of BFU-E from either bone marrow or peripheral blood. It had little effect on the colony formation of CFU- GM and the mixed colonies from CFU-GEMM. In serum-depleted culture, the effect of activin was shown to be dose-dependent with doses effective at picomolar concentrations. The potentiation effect of activin was exerted indirectly through mediation of both monocytes and T lymphocytes. Activin was also found to increase specifically the proportion of DNA-synthesizing erythroid progenitors from both bone marrow and peripheral blood. It had little effect on DNA synthesis in CFU-GM and in mitogen-stimulated lymphocytes. Addition of the monocytes or T lymphocytes to their respective depleted subpopulations of mononuclear cells reconstituted the enhancing effect of activin on the colony formation and DNA synthesis of erythroid progenitors. These results strongly suggest a specific role of activin in potentiating the proliferation and differentiation of erythroid progenitors in vitro.

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