scholarly journals Relationship between in vitro colony-forming ability and expression of Ia-like antigen of acute nonlymphocytic leukemia (ANLL) cells

Blood ◽  
1980 ◽  
Vol 56 (5) ◽  
pp. 859-865 ◽  
Author(s):  
A Hiraoka ◽  
M Yamagishi ◽  
T Ohkubo ◽  
Y Yoshida ◽  
H Uchino
Keyword(s):  
Bone Marrow Cells ◽  
Type A ◽  
Colony Growth ◽  
Leukemic Cells ◽  
Acute Nonlymphocytic Leukemia ◽  
Functional Maturation ◽  
Type C ◽  
Differentiation And Proliferation ◽  
Stimulating Factor

Abstract In vitro colony-forming ability of untreated acute nonlymphocytic leukemia (ANLL) cells determined by the CFU-C assay with and without PHA presensitization, and also human Ia-like antigen of their surfaces were investigated. In vitro colony-forming ability of 32 ANLLs was classified into four types: (A) no colony growth: 16, (B) PHA-dependent colony growth: 9, (C) colony-stimulating factor (CSF) dependent colony growth: 5, and (D) CSF and PHA independent colony growth: 2. The pattern of 12 normal bone marrow cells corresponded to type C. Leukemic cells were Ia-positive in type A, negative or weakly positive in type B, and negative in type C. Colonies as seen in types B and C consisted mostly of monocytes-macrophages with myeloid cells at varyi stages of maturation up to metamyelocyte. Incubation of type A ANLL cells with anti-Ia-serum followed by PHA presensitization resulted in development of colonies in some analyzed cases with moderately positive Ia-like antigen. Correlation of in vitro colony-forming ability with the expression of human Ia-like antigen of ANLL cells indicated not only a hematologic heterogeneity of ANLL in differentiation and proliferation but also biologic significance of Ia-like antigen in reference to functional maturation of ANLL cells.

scholarly journals Relationship between in vitro colony-forming ability and expression of Ia-like antigen of acute nonlymphocytic leukemia (ANLL) cells

Blood ◽  
1980 ◽  
Vol 56 (5) ◽  
pp. 859-865
Author(s):  
A Hiraoka ◽  
M Yamagishi ◽  
T Ohkubo ◽  
Y Yoshida ◽  
H Uchino
Keyword(s):  
Bone Marrow Cells ◽  
Type A ◽  
Colony Growth ◽  
Leukemic Cells ◽  
Acute Nonlymphocytic Leukemia ◽  
Functional Maturation ◽  
Type C ◽  
Differentiation And Proliferation ◽  
Stimulating Factor

In vitro colony-forming ability of untreated acute nonlymphocytic leukemia (ANLL) cells determined by the CFU-C assay with and without PHA presensitization, and also human Ia-like antigen of their surfaces were investigated. In vitro colony-forming ability of 32 ANLLs was classified into four types: (A) no colony growth: 16, (B) PHA-dependent colony growth: 9, (C) colony-stimulating factor (CSF) dependent colony growth: 5, and (D) CSF and PHA independent colony growth: 2. The pattern of 12 normal bone marrow cells corresponded to type C. Leukemic cells were Ia-positive in type A, negative or weakly positive in type B, and negative in type C. Colonies as seen in types B and C consisted mostly of monocytes-macrophages with myeloid cells at varyi stages of maturation up to metamyelocyte. Incubation of type A ANLL cells with anti-Ia-serum followed by PHA presensitization resulted in development of colonies in some analyzed cases with moderately positive Ia-like antigen. Correlation of in vitro colony-forming ability with the expression of human Ia-like antigen of ANLL cells indicated not only a hematologic heterogeneity of ANLL in differentiation and proliferation but also biologic significance of Ia-like antigen in reference to functional maturation of ANLL cells.

scholarly journals Colony formation in vitro by leukemic cells in acute lymphoblastic leukemia (ALL)

Blood ◽  
1978 ◽  
Vol 52 (4) ◽  
pp. 712-718 ◽  
Author(s):  
SD Smith ◽  
EM Uyeki ◽  
JT Lowman
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
Bone Marrow Cells ◽  
Lymphoblastic Leukemia ◽  
Lymphoid Cells ◽  
Assay System ◽  
Leukemic Cells ◽  
Specific Cell ◽  
Specific Cell Surface

Abstract An assay system in vitro for the growth of malignant lymphoblastic colony-forming cells (CFC) was established. Growth of malignant myeloblastic CFC has been previously reported, but this is the first report of growth of malignant lymphoblastic CFC. Established assay systems in vitro have been very helpful in elucidating the control of growth and differentiation of both normal and malignant bone marrow cells. Lymphoblastic CFC were grown from the bone marrow aspirates of 20 children with acute lymphoblastic leukemia. Growth of these colonies was established on an agar assay system and maintained in the relative hypoxia (7% oxygen) of a Stulberg chamber. The criteria for malignancy of these colonies was based upon cellular cytochemical staining characteristics, the presence of specific cell surface markers, and the ability of these lymphoid cells to grow without the addition of a lymphoid mitogen. With this technique, specific nutritional requirements and drug sensitivities can be established in vitro, and these data may permit tailoring of individual antileukemic therapy.

scholarly journals In vitro interactions of PGE and cAMP with murine and human erythroid precursors

Blood ◽  
1980 ◽  
Vol 56 (1) ◽  
pp. 74-79 ◽  
Author(s):  
GB Rossi ◽  
AR Migliaccio ◽  
G Migliaccio ◽  
F Lettieri ◽  
M Di Rosa ◽  
...  
Keyword(s):  
Inhibitory Action ◽  
Colony Growth ◽  
Inhibitory Influence ◽  
Adherent Cells ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent ◽  
Stimulating Factor ◽  
In Vitro Interactions

Abstract Addition of prostaglandins of the E series (PGE1, PGE2) in methylcellulose cultures of murine marrow results in a dose-dependent inhibition of the cloning efficiency of both BFU-E and CFU-C. However, CFU-E growth is unaffected. The inhibitory action of PGE is progressively overcome by increasing amounts of colony-stimulating factor (CSF), and with some limitations, also of erythropoietin (Ep). Addition of PGF2 alpha' associated or not with indomethacin, does not exert any significant effect on these hemopoietic precursors. In an attempt to unvail the mechanism(s) underlying these phenomena, dibutyryl-cyclic AMP (db-cAMP), theophylline (an inhibitor of phosphodiesterase), or theophylline + PGE were plated at various concentrations. Both db-cAMP and theophylline induce an inhibitory influence on both BFU-E and CFU-C growth, which mimicks that by PGEs; additionally, theophylline potentiates the inhibitory action of PGE1. In all these studies, the CFU-E number was not significantly modified. PGE action on BFU-E proliferation is clearly species-dependent, since PGE1 addition to human marrow methylcellulose cultures induces a significant enhancement of the number of both BFU-E and CFU-E derived colonies. This action was abolished upon removal of adherent cells, thus suggesting that PGE1 evokes a release of factor(s) enhancing human erythroid colony growth by adherent cells.

scholarly journals In vitro restoration of polyclonal hematopoiesis in a chronic myelogenous leukemia after in vitro treatment with 4- hydroperoxycyclophosphamide

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 753-757 ◽  
Author(s):  
G Degliantoni ◽  
L Mangoni ◽  
V Rizzoli
Keyword(s):  
Bone Marrow ◽  
Chronic Myelogenous Leukemia ◽  
Early Phase ◽  
Bone Marrow Cells ◽  
Type A ◽  
G6pd Activity ◽  
Myelogenous Leukemia ◽  
Stem Cell Population ◽  
Marrow Cells

Bone marrow cells of a 45-year-old female with Philadelphia chromosome (Ph1)-positive, early-phase chronic myelogenous leukemia (CML), who was heterozygous for the glucose-6-phosphate dehydrogenase (G6PD) locus, were pretreated in vitro with 4-hydroperoxycyclophosphamide (4-HC) and tested for G6PD activity in several colony formation assays and for karyotypic abnormalities. All cells within the mixed (CFU-GEMM), the erythroid burst (BFU-E), and the granulocyte-macrophage (CFU-GM) colonies expressed type A and type B G6PD activity and a normal karyotype, whereas untreated cells expressed type A G6PD and the Ph1 chromosome. This reversal of G6PD activity type and the disappearance of the Ph1 chromosome in colonies grown from 4-HC-treated cells indicate that this cytotoxic agent spares a residual normal stem cell population in bone marrow cells of early-phase CML patients. This finding, in turn, suggests a therapeutic approach in CML based on in vitro chemotherapy of autologous bone marrow grafts.

scholarly journals Interleukin-6 enhances murine megakaryocytopoiesis in serum-free culture

Blood ◽  
1990 ◽  
Vol 75 (12) ◽  
pp. 2286-2291 ◽  
Author(s):  
K Koike ◽  
T Nakahata ◽  
T Kubo ◽  
T Kikuchi ◽  
M Takagi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Interleukin 6 ◽  
Bone Marrow Cells ◽  
Early Stage ◽  
Colony Growth ◽  
Colony Stimulating Factor ◽  
Serum Free ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Marrow Cells

We investigated the effect of interleukin-6 (IL-6) on murine megakaryocytopoiesis in a serum-free culture system. The addition of IL- 6 to a culture containing interleukin-3 (IL-3) resulted in a significant increase in the number of megakaryocyte colonies by bone marrow cells of normal mice. The megakaryocytic progenitors that survive exposure to 5-fluorouracil (5-FU) exhibited a more significant response to IL-6 and IL-3. Polyclonal anti-IL-6 antibody neutralized the stimulatory effect of IL-6 on megakaryocyte colony growth supported by IL-3. Delayed addition experiments and replating experiments of blast cell colonies showed that megakaryocytic progenitors are supported by IL-3 in the early stage of the development but require IL- 6 for their subsequent proliferation and differentiation. In addition, IL-6 increased the size of megakaryocytes in granulocyte-macrophage- megakaryocyte colonies. The combination of granulocyte colony- stimulating factor or granulocyte-macrophage colony stimulating factor with IL-3 resulted in an increase in the granulocyte-macrophage colony growth of bone marrow cells of 5-FU-treated mice or normal mice, respectively, but had little effect on the enhancement of pure and mixed megakaryocyte colony growth. These results suggest that IL-6 plays an important role in murine megakaryocytopoiesis.

scholarly journals Clonal analysis of basophil differentiation in bone marrow cultures from a Down's syndrome patient with megakaryoblastic leukemia

Blood ◽  
1985 ◽  
Vol 66 (6) ◽  
pp. 1278-1283
Author(s):  
T Suda ◽  
J Suda ◽  
Y Miura ◽  
Y Hayashi ◽  
M Eguchi ◽  
...  
Keyword(s):  
Single Cells ◽  
Down's Syndrome ◽  
Calcium Ionophore ◽  
Down’S Syndrome ◽  
Colony Growth ◽  
Leukemic Cells ◽  
Cytologic Examination ◽  
Megakaryoblastic Leukemia

We present the in vitro differentiation of marrow cells from a patient with Down's syndrome accompanied by megakaryoblastic leukemia into basophils in the presence of phytohemagglutinin-stimulated leukocyte conditioned medium, using a liquid culture and methylcellulose culture system. Identification of basophils was established by metachromatic staining with toluidine blue, transmission electron microscopy, and the presence of histamine. However, these basophils did not release histamine in response to calcium ionophore or chemotactic peptide. Samples from suspension cultures that contained 90% basophils showed chromosomal markers characteristic of leukemic cells (48, XY, +11, +21, t(1;15)) in all examined mitoses. The cellular composition of leukemic colonies grown in methylcellulose culture from single cells was studied using the micromanipulation technique. High plating efficiency and extreme predominance of basophil colonies were observed. In a total 137 cultures, 79 revealed colony growth. Of 59 colonies that were analyzed by cytologic examination, 46 were pure basophil colonies. These basophil colonies showed disperse morphology, similar to that of a normal basophil colony. The clonality of the basophil colonies and skewing of lineage expression were documented from leukemic single-cell cultures. These data showed that leukemic cells have the capacity for differentiation into some lineages that are not expressed in vivo.

scholarly journals Interleukin-6 (IL-6) is an intermediate in IL-1-induced proliferation of leukemic human megakaryoblasts

Blood ◽  
1990 ◽  
Vol 76 (10) ◽  
pp. 1972-1979 ◽  
Author(s):  
MA Brach ◽  
B Lowenberg ◽  
L Mantovani ◽  
U Schwulera ◽  
R Mertelsmann ◽  
...  
Keyword(s):  
Thymidine Incorporation ◽  
Interleukin 1 ◽  
Leukemic Cells ◽  
Megakaryoblastic Leukemia ◽  
Gm Csf ◽  
Endogenous Production ◽  
Neutralizing Monoclonal Antibody ◽  
In Vitro Effects ◽  
Stimulating Factor

Abstract We have examined the in vitro effects of recombinant human (rh) interleukin-1 (IL-1) on the growth of purified megakaryoblasts obtained from patients with acute megakaryoblastic leukemia. We demonstrate that both IL-1 alpha and IL-1 beta treatment of these cells led to stimulation of DNA synthesis (as shown by increase of 3H-thymidine incorporation up to 35-fold) and also resulted in colony formation of leukemic megakaryoblasts. However, the stimulatory effect of IL-1 was dependent on endogenous production of IL-6, because addition of neutralizing monoclonal antibody (MoAb) to IL-6 abrogated the stimulatory activity of IL-1. In contrast, neutralizing MoAbs to granulocyte (G)-colony stimulating factor (CSF), granulocyte-macrophage (GM)-CSF, and macrophage (M)-CSF failed to counteract the growth- enhancing effects of IL-1. Leukemic megakaryoblasts accumulated IL-6 mRNA and released IL-6 protein into their culture supernatant when exposed to rh IL-1 but failed to disclose transcripts for G-, GM-, and M-CSF under these conditions. Analysis of IL-6 receptor (IL-6R) transcript levels demonstrated that megakaryoblasts constitutively expressed IL-6R mRNA and that these transcripts are down-regulated to undetectable levels upon exposure to IL-1 and IL-6. Increase of 3H- thymidine incorporation by megakaryoblasts could be duplicated by exogenous IL-6 that could be blocked by neutralizing MoAb to IL-6. In conclusion, our results suggest that leukemic megakaryoblasts could produce and secrete IL-6, and express IL-6R, and that the growth- enhancing effect of IL-1 on these cells is indirect, via production of IL-6 by leukemic cells.

scholarly journals Enhanced expression of the granulocyte-macrophage colony stimulating factor gene in acute myelocytic leukemia cells following in vitro blast cell enrichment

Blood ◽  
1988 ◽  
Vol 72 (4) ◽  
pp. 1329-1332 ◽  
Author(s):  
DC Kaufman ◽  
MR Baer ◽  
XZ Gao ◽  
ZQ Wang ◽  
HD Preisler
Keyword(s):  
T Cell ◽  
Leukemic Cells ◽  
Acute Myelocytic Leukemia ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Myelocytic Leukemia ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Expression of the granulocyte-macrophage colony-stimulating factor (GM- CSF) gene in acute myelocytic leukemia (AML) was assayed by Northern blot analysis. GM-CSF messenger RNA (mRNA) was detected in the freshly obtained mononuclear cells of only one of 48 cases of AML, in contrast with recent reports that GM-CSF mRNA might be detected in half of the cases of AML when RNA is prepared from T-cell- and monocyte-depleted leukemic cells. We did find, however, that expression of the GM-CSF gene was detectable in five of ten cases after in vitro T-cell and monocyte depletion steps. Additional studies suggest that expression of GM-CSF in the bone marrow of the one positive case, rather than being autonomous, was under exogenous control, possibly by a paracrine factor secreted by marrow stromal cells. These studies emphasize the potential for altering in vivo patterns of gene expression by in vitro cell manipulation.

scholarly journals Effects of novel retinoid X receptor-selective ligands on myeloid leukemia differentiation and proliferation in vitro

Blood ◽  
1996 ◽  
Vol 87 (5) ◽  
pp. 1977-1984 ◽  
Author(s):  
M Kizaki ◽  
MI Dawson ◽  
R Heyman ◽  
E Elster ◽  
R Morosetti ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Clonal Growth ◽  
Myeloid Leukemia ◽  
Target Genes ◽  
Synergistic Effects ◽  
Leukemic Cells ◽  
Acute Myeloid Leukemia Cells ◽  
Differentiation And Proliferation ◽  
Acute Myeloid

The biologic effects of retinoids such as all-trans-retinoic acid (ATRA) and 9-cis-retinoic acid on proliferation and differentiation of hematopoietic cells are mediated by binding and activating two distinct families of transcription factors: the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs). The RARs require heterodimerization with RXRs; in addition, RXRs can form homodimers, which can bind to DNA response elements that are either distinct or the same as those bound by the RAR/RXR heterodimers. Therefore, the two retinoid pathways provide sequences that are specific for effective DNA binding and activation of target genes. We have developed several series of novel synthetic retinoids that selectively interact with RXR/RXR homodimers and RAR/RXR heterodimers. We show here that SR11236 and SR11246, which are RXR-selective analogs, had little ability to inhibit clonal growth and induce differentiation of leukemic cells (HL- 60 cells and fresh acute myeloid leukemia cells). However, SR11249, SR11256, and LGD1069, which activated both RXR/RXR homodimers and RAR/RXR heterodimers, could inhibit clonal growth and induce differentiation of HL-60 cells as well as leukemic cells from patients, including those with acute promyelocytic leukemia (APL). This is similar to results observed with RAR/RXR-specific ligands. Interestingly, the combination of ATRA and either SR11249, SR11256, or LGD1069 showed synergistic effects in inducing differentiation of HL-60 cells. A retinoid (SR11238) with strong anti-AP-1 activity that did not activate the RARs and RXRs for gene transcription from the response element TREpal was inactive in our assay systems, suggesting that the antiproliferative effects of retinoids on leukemic cells is not mediated by inhibiting the AP-1 pathway. We conclude that the RAR/RXR pathway is more important than RXR/RXR pathway for differentiation and proliferation of acute myeloid leukemic cells, and certain retinoids or combination of retinoids with both RAR and RXR specificities may synergistically enhance the differentiation activity of ATRA, which may be relevant in several clinical situations.

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