Histochemical Demonstration of Phosphorylase in Blood and Bone Marrow Cells

Abstract Phosphorylase activity has been demonstrated histochemically in blood and bone marrow cells. The polysaccharide newly synthesized from glucose-1-phosphate by the enzyme of these cells gave a blue or violet blue iodine staining reaction. The reaction occurred in cytoplasm and the nuclei did not stain. This was shown in the neutrophilic, eosinophilic or pseudoeosinophilic leukocytes of man, dog, rabbit and pigeon peripheral blood. It was likewise demonstrated in the bicolored leukocytes of snake, fish and frog. The reaction also appeared in the neutrophilic, pseudoeosinophilic or eosinophilic leukocytes, metamyelocytes, myelocytes and promyelocytes of human and rabbit bone marrow smears.

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Manifestation of Pig-a mutant bone marrow erythroids and peripheral blood erythrocytes in mice treated with N-ethyl-N-nitrosourea: Direct sequencing of Pig-a cDNA from bone marrow cells negative for GPI-anchored protein expression

Mutation Research/Genetic Toxicology and Environmental Mutagenesis ◽

10.1016/j.mrgentox.2011.03.016 ◽

2011 ◽

Vol 723 (1) ◽

pp. 36-42 ◽

Cited By ~ 46

Author(s):

Takafumi Kimoto ◽

Kumiko Suzuki ◽

Xiao mei Kobayashi ◽

Vasily N. Dobrovolsky ◽

Robert H. Heflich ◽

...

Keyword(s):

Bone Marrow ◽

Protein Expression ◽

Peripheral Blood ◽

Bone Marrow Cells ◽

Direct Sequencing ◽

Marrow Cells

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Adult Mice With Targeted Mutation of the Interleukin-11 Receptor (IL11Ra) Display Normal Hematopoiesis

Blood ◽

10.1182/blood.v90.6.2148 ◽

1997 ◽

Vol 90 (6) ◽

pp. 2148-2159 ◽

Cited By ~ 118

Author(s):

Harshal H. Nandurkar ◽

Lorraine Robb ◽

David Tarlinton ◽

Louise Barnett ◽

Frank Köntgen ◽

...

Keyword(s):

Bone Marrow ◽

Peripheral Blood ◽

Blood Cells ◽

Bone Marrow Cells ◽

White Blood Cells ◽

Null Mutation ◽

Wild Type ◽

Normal Numbers ◽

Interleukin 11 ◽

Marrow Cells

Abstract Interleukin-11 (IL-11) is a pleiotropic growth factor with a prominent effect on megakaryopoiesis and thrombopoiesis. The receptor for IL-11 is a heterodimer of the signal transduction unit gp130 and a specific receptor component, the α-chain (IL-11Rα). Two genes potentially encode the IL-11Rα: the IL11Ra and IL11Ra2 genes. The IL11Ra gene is widely expressed in hematopoietic and other organs, whereas the IL11Ra2 gene is restricted to only some strains of mice and its expression is confined to testis, lymph node, and thymus. To investigate the essential actions mediated by the IL-11Rα, we have generated mice with a null mutation of IL11Ra (IL11Ra−/−) by gene targeting. Analysis of IL11Ra expression by Northern blot and reverse transcriptase-polymerase chain reaction, as well as the absence of response of IL11Ra−/− bone marrow cells to IL-11 in hematopoietic assays, further confirmed the null mutation. Compensatory expression of the IL11Ra2 in bone marrow cells was not detected. IL11Ra−/− mice were healthy with normal numbers of peripheral blood white blood cells, hematocrit, and platelets. Bone marrow and spleen contained normal numbers of cells of all hematopoietic lineages, including megakaryocytes. Clonal cultures did not identify any perturbation of granulocyte-macrophage (GM), erythroid, or megakaryocyte progenitors. The number of day-12 colony-forming unit-spleen progenitors were similar in wild-type and IL11Ra−/− mice. The kinetics of recovery of peripheral blood white blood cells, platelets, and bone marrow GM progenitors after treatment with 5-flurouracil were the same in IL11Ra−/− and wild-type mice. Acute hemolytic stress was induced by phenylhydrazine and resulted in a 50% decrease in hematocrit. The recovery of hematocrit was comparable in IL11Ra−/− and wild-type mice. These observations indicate that IL-11 receptor signalling is dispensable for adult hematopoiesis.

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Analysis of interferon-inducible genes in cells of chronic myeloid leukemia patients responsive or resistant to an interferon-alpha treatment

Blood ◽

10.1182/blood.v76.11.2337.bloodjournal76112337 ◽

1990 ◽

Vol 76 (11) ◽

pp. 2337-2342

Author(s):

IM Clauss ◽

B Vandenplas ◽

MG Wathelet ◽

C Dorval ◽

A Delforge ◽

...

Keyword(s):

Bone Marrow ◽

Chronic Myeloid Leukemia ◽

Interferon Alpha ◽

Peripheral Blood ◽

Myeloid Leukemia ◽

Bone Marrow Cells ◽

Healthy Controls ◽

Ifn Alpha ◽

Inducible Genes ◽

Marrow Cells

Recombinant human interferon-alpha (IFN-alpha) can induce a hematologic remission in patients with chronic myeloid leukemia. However, some patients are resistant and others develop late resistance to the IFN- alpha treatment. To understand the molecular mechanism of this resistance, we have analyzed the expression of 10 IFN-inducible genes in the cells of three resistant patients, two responsive patients, and six healthy controls. Northern blot hybridizations showed that all the genes were induced in in vitro IFN-alpha treated peripheral blood cells of the patients and healthy controls. These genes were also inducible in peripheral blood and bone marrow cells of two out of two resistant patients administered an injection of IFN-alpha. We conclude that the resistance to the IFN-alpha treatment of the chronic myeloid leukemia patients we studied is not due to (1) the absence of induction of any of the 10 IFN-inducible genes we studied, including the low-molecular- weight 2′-5′oligoadenylate synthetase; (2) the presence of an antagonist of IFN-alpha in the peripheral blood or bone marrow cells; and (3) the presence of neutralizing anti-IFN-alpha antibodies.

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5-Azacytidine increases gamma-globin synthesis and reduces the proportion of dense cells in patients with sickle cell anemia

Blood ◽

10.1182/blood.v62.2.370.370 ◽

1983 ◽

Vol 62 (2) ◽

pp. 370-380 ◽

Cited By ~ 37

Author(s):

TJ Ley ◽

J DeSimone ◽

CT Noguchi ◽

PH Turner ◽

AN Schechter ◽

...

Keyword(s):

Bone Marrow ◽

Sickle Cell ◽

Sickle Cell Anemia ◽

Peripheral Blood ◽

Bone Marrow Cells ◽

Beta Thalassemia ◽

Globin Genes ◽

Gamma Globin ◽

Globin Synthesis ◽

Marrow Cells

Abstract We previously demonstrated that 5-azacytidine can selectively increase gamma-globin synthesis in a patient with beta +-thalassemia, prompting us to treat two patients with sickle cell anemia and two additional patients with beta + thalassemia. 5-Azacytidine (2 mg/kg/day) was continuously infused for 7 days with no apparent clinical toxicity. The gamma/beta-globin biosynthetic ratio increased fourfold to sixfold in the bone marrow cells of each patient after treatment and remained elevated for 7–14 additional days. Hypomethylation of DNA near the gamma-globin genes in bone marrow cells was demonstrated 2 days after beginning the 5-azacytidine infusion. The peripheral blood fetal hemoglobin (HbF) level increased from 6.0% to 13.7% in one patient with sickle cell anemia and from 1.6% to 8.9% in the second. Stractan gradient analyses of peripheral blood from patients with sickle cell anemia revealed a marked decrease in the percentage of dense cells (cells that contain increased amounts of HbS polymer when deoxygenated) following treatment. These observations provide an impetus to investigate the effects of repeated courses of 5-azacytidine in a small group of severely ill patients to determine whether this drug may have a role in the treatment of patients with sickle cell anemia and beta- thalassemia.

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Detection by enzymatic amplification of bcr-abl mRNA in peripheral blood and bone marrow cells of patients with chronic myelogenous leukemia

Blood ◽

10.1182/blood.v73.6.1735.1735 ◽

1989 ◽

Vol 73 (6) ◽

pp. 1735-1741 ◽

Cited By ~ 44

Author(s):

W Lange ◽

DS Snyder ◽

R Castro ◽

JJ Rossi ◽

KG Blume

Keyword(s):

Bone Marrow ◽

Chronic Myelogenous Leukemia ◽

Peripheral Blood ◽

Bone Marrow Cells ◽

Philadelphia Chromosome ◽

Chromosome 9 ◽

Myelogenous Leukemia ◽

Enzymatic Amplification ◽

Polymerase Chain ◽

Marrow Cells

Abstract The Philadelphia chromosome of chronic myelogenous leukemia (CML) patients is caused by a translocation of the c-abl gene from chromosome 9 to the breakpoint cluster region (bcr) on chromosome 22. A new bcr- abl mRNA is expressed in these cases. We have developed a modified polymerase chain reaction (PCR) for the detection of this mRNA. The method is extremely sensitive, reliable, and relatively fast. The analysis of peripheral blood or bone marrow cells from CML patients treated with chemotherapy shows that the two possible mRNAs are expressed in various combinations. Our results show that even after myeloablative therapy for bone marrow transplantation bcr-abl mRNAs are still expressed. Further studies, however, are necessary to determine the clinical relevance of a small number of persisting cells expressing the bcr-abl mRNA.

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Long-term multilineage expression in peripheral blood from a Moloney murine leukemia virus vector after serial transplantation of transduced bone marrow cells

Blood ◽

10.1182/blood.v95.3.829.003k18_829_836 ◽

2000 ◽

Vol 95 (3) ◽

pp. 829-836 ◽

Cited By ~ 8

Author(s):

Timothy W. Austin ◽

Suzan Salimi ◽

Gabor Veres ◽

Franck Morel ◽

Heini Ilves ◽

...

Keyword(s):

Bone Marrow ◽

Peripheral Blood ◽

Blood Cells ◽

Transgene Expression ◽

Bone Marrow Cells ◽

Leukemia Virus ◽

Moloney Murine Leukemia Virus ◽

Marrow Cells

Using a mouse bone marrow transplantation model, the authors evaluated a Moloney murine leukemia virus (MMLV)-based vector encoding 2 anti-human immunodeficiency virus genes for long-term expression in blood cells. The vector also encoded the human nerve growth factor receptor (NGFR) to serve as a cell-surface marker for in vivo tracking of transduced cells. NGFR+ cells were detected in blood leukocytes of all mice (n=16; range 16%-45%) 4 to 5 weeks after transplantation and were repeatedly detected in blood erythrocytes, platelets, monocytes, granulocytes, T cells, and B cells of all mice for up to 8 months. Transgene expression in individual mice was not blocked in the various cell lineages of the peripheral blood and spleen, in several stages of T-cell maturation in the thymus, or in the Lin−/loSca-1+ and c-kit+Sca-1+ subsets of bone marrow cells highly enriched for long-term multilineage-reconstituting activity. Serial transplantation of purified NGFR+c-kit+Sca-1+bone marrow cells resulted in the reconstitution of multilineage hematopoiesis by donor type NGFR+ cells in all engrafted mice. The authors concluded that MMLV-based vectors were capable of efficient and sustained transgene expression in multiple lineages of peripheral blood cells and hematopoietic organs and in hematopoietic stem cell (HSC) populations. Differentiation of engrafting HSC to peripheral blood cells is not necessarily associated with dramatic suppression of retroviral gene expression. In light of earlier studies showing that vector elements other than the long-terminal repeat enhancer, promoter, and primer binding site can have an impact on long-term transgene expression, these findings accentuate the importance of empirically testing retroviral vectors to determine lasting in vivo expression.

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