Differential Regulation of Cell Membrane Receptors for Interferon-γ and -α on Peripheral Blood Cells of Patients with Chronic Myelogenous Leukemia During Interferon-γ/α Therapy

1988 ◽  
pp. 63-73
Author(s):  
H. H. Bartsch ◽  
U. Ücer ◽  
G. A. Nagel ◽  
K. Pfizenmaier
Keyword(s):  
Cell Membrane ◽  
Chronic Myelogenous Leukemia ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Interferon Γ ◽  
Differential Regulation ◽  
Membrane Receptors ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells

scholarly journals BCR-ABL protein expression in peripheral blood cells of chronic myelogenous leukemia patients undergoing therapy

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3629-3637 ◽  
Author(s):  
JQ Guo ◽  
JY Lian ◽  
YM Xian ◽  
MS Lee ◽  
AB Deisseroth ◽  
...  
Keyword(s):  
Protein Expression ◽  
Chronic Myelogenous Leukemia ◽  
Western Blotting ◽  
Peripheral Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Philadelphia Chromosome ◽  
Chronic Phase ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells

Chronic myelogenous leukemia (CML) is a myeloproliferative disorder associated with the Philadelphia chromosome (Ph1) in more than 95% of these patients. The Ph1 and the resulting BCR-ABL fused genes are markers for this type of leukemia. In CML, the product of the fused BCR- ABL gene is typically a protein of approximately 2,000 amino acids termed P210 BCR-ABL. We have developed an assay for the BCR-ABL protein involving Western blotting of circulating white blood cells (WBC) with an anti-ABL monoclonal antibody that can detect P210 BCR-ABL and P145 ABL in peripheral blood cells from chronic phase Ph1-positive leukemia patients. This assay was used to analyze the BCR-ABL protein content of circulating WBC from CML patients before and after various treatments. In parallel to changes in percentages of Ph1-positive blood cells as determined by cytogenetic analyses of bone marrow samples, BCR-ABL protein expression in blood cells decreased or increased as patients entered remission or underwent relapse. Of interest, six Ph1-negative CML patients were BCR-ABL protein-positive. All except one had a rearrangement in the major breakpoint cluster region and that patient expressed P185 BCR-ABL and not P210. Our results indicate that the BCR- ABL Western blotting assay has clinical applications for both diagnosis and prospective evaluation of Ph1-positive and Ph1-negative CML patients.

scholarly journals BCR-ABL protein expression in peripheral blood cells of chronic myelogenous leukemia patients undergoing therapy

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3629-3637 ◽  
Author(s):  
JQ Guo ◽  
JY Lian ◽  
YM Xian ◽  
MS Lee ◽  
AB Deisseroth ◽  
...  
Keyword(s):  
Protein Expression ◽  
Chronic Myelogenous Leukemia ◽  
Western Blotting ◽  
Peripheral Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Philadelphia Chromosome ◽  
Chronic Phase ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells

Abstract Chronic myelogenous leukemia (CML) is a myeloproliferative disorder associated with the Philadelphia chromosome (Ph1) in more than 95% of these patients. The Ph1 and the resulting BCR-ABL fused genes are markers for this type of leukemia. In CML, the product of the fused BCR- ABL gene is typically a protein of approximately 2,000 amino acids termed P210 BCR-ABL. We have developed an assay for the BCR-ABL protein involving Western blotting of circulating white blood cells (WBC) with an anti-ABL monoclonal antibody that can detect P210 BCR-ABL and P145 ABL in peripheral blood cells from chronic phase Ph1-positive leukemia patients. This assay was used to analyze the BCR-ABL protein content of circulating WBC from CML patients before and after various treatments. In parallel to changes in percentages of Ph1-positive blood cells as determined by cytogenetic analyses of bone marrow samples, BCR-ABL protein expression in blood cells decreased or increased as patients entered remission or underwent relapse. Of interest, six Ph1-negative CML patients were BCR-ABL protein-positive. All except one had a rearrangement in the major breakpoint cluster region and that patient expressed P185 BCR-ABL and not P210. Our results indicate that the BCR- ABL Western blotting assay has clinical applications for both diagnosis and prospective evaluation of Ph1-positive and Ph1-negative CML patients.

scholarly journals Impact of Short-Term Systemic Hypoxia on Phagocytosis, Cytokine Production, and Transcription Factor Activation in Peripheral Blood Cells

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Michael Fritzenwanger ◽  
Christian Jung ◽  
Bjoern Goebel ◽  
Alexander Lauten ◽  
Hans R. Figulla
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Cytokine Production ◽  
Interferon Γ ◽  
Peripheral Blood Cells ◽  
Immune Functions ◽  
Cellular Functions ◽  
Short Term ◽  
Transcription Factor Activation ◽  
Systemic Hypoxia

Hypoxia frequently associated with certain physiologic and pathologic conditions influences numerous cellular functions. Because the effects of short-term hypoxia are incompletely understood, we examined phagocytosis and cytokine production as well as the activation of the transcription factors HIF-1 and NFκB in peripheral blood cells of healthy volunteers exposed to an oxygen concentration equivalent to that found at a height of 5500 m. Furthermore, we analysed plasma HIF-1 and serum concentrations of various HIF-1-dependent genes. Results showed that short-term hypoxia increased phagocytosis in neutrophils without affecting monocyte phagocytosis. Hypoxia decreased basal TNFα concentration in monocytes and basal interferon γ concentration in CD4+T lymphocytes. In contrast, plasma HIF and serum VEGF concentrations were not affected by hypoxia, although serum EPO concentration was raised. In PBMC, hypoxia increased cytosolic HIF-1 concentration without affecting nuclear HIF-1 concentration and led to a rise in the nuclear NFκB in PBMC. Our results show that short-term hypoxia affects immune functions in healthy individuals. Furthermore, we speculate that the effects of hypoxia are not due to HIF-1, but are caused by the activation of NFκB .

scholarly journals Chromosome Studies in Normal and Leukemic Rats

Blood ◽  
1964 ◽  
Vol 23 (5) ◽  
pp. 564-571 ◽  
Author(s):  
G. DOWD ◽  
K. DUNN ◽  
WILLIAM C. MOLONEY
Keyword(s):  
Bone Marrow ◽  
Blood Cell ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Species Differences ◽  
Peripheral Blood Cell ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells ◽  
X Ray ◽  
Normal Rat

Abstract 1. Adequate chromosome preparations were obtained in 70 per cent of normal rat peripheral blood cell cultures. However, cultures of peripheral blood cells from leukemic rats were almost universally unsuccessful. 2. In x-ray- and 3MCA-induced leukemias direct bone marrow preparations provided adequate metaphases in eight of 12 cases. Failures were attributed in four cases to scanty material obtained from fibrotic marrows. 3. No consistent chromosome abnormalities, such as those reported in human myelogenous leukemia, were found in these leukemic rats. However, the series of cases is small, and species differences and other factors may have influenced the results of these studies.

A Distinctive Pattern of Different Gene Expression in Chronic Myelogenous Leukemia Patients

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4231-4231
Author(s):  
Carla Di Stefano ◽  
Marco Tafani ◽  
Bruna Pucci ◽  
Elisabetta Abruzzese ◽  
Margherita Trawiska ◽  
...  
Keyword(s):  
Western Blot ◽  
Chronic Myelogenous Leukemia ◽  
Molecular Chaperones ◽  
Blot Analysis ◽  
Blood Cells ◽  
Western Blot Analysis ◽  
Blast Crisis ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells ◽  
Imatinib Resistant

Abstract Introduction: Molecular chaperones have many functions, such as protecting other proteins against aggregation, assisting in folding of nascent proteins/refolding of damaged proteins and targeting severely damaged proteins to degradation. As one of the molecular chaperones, Hsp90 functions to facilitate the folding of newly synthesized and denatured client proteins, including mutated p53, Bcr-Abl, p185ErbB2 and Raf-1. The Bcr-Abl fusion gene encodes for the p210Bcr-Abl tyrosine kinase (TK) implicated in the pathogenesis of chronic myelogenous leukemia (CML). Studies in cultured cells have identified many signal transduction pathways activated by Bcr-Abl, including activation of the Ras, MAPK, JNK/SAPK, phosphatidylinositol-3 kinase, nuclear factor-B and STAT pathways. Imatinib mesylate (imatinib IM) is a tyrosine kinase inhibitor that competitively inhibits ATP binding in the kinase domains of both the Bcr-Abl and c-Abl kinases. It has been suggested that resistance to imatinib stems from Bcr-Abl gene amplification, leading to overexpression of Bcr-Abl protein or point mutations in the Bcr-Abl gene However, several groups suggested that there might be other forms of Bcr-Abl-independent imatinib resistance Recently, it has been reported that changes in histone deacetylase (HDAC) expression in leukemic cells could be involved in mechanisms for abnormal cellular proliferation that operate through chromatin-independent pathways and thereby could lead to acquired drug resistance of the cells In the present study, we evaluated in primary leukemic blasts, obtained from chronic myelogenous leukemia patients at onset, patients in blast crisis and patients which were imatinib-resistant The espression the sirtuin members family and HSP70, HSP90 i-NOS and bcl-2 was evaluated by Nortern blot and Western blot analysis. Material and Methods: Primary leukaemia blasts We harvested primary blast rich mononuclear cells were obtained by gradient centrifugation on ficoll-hypaque of bone marrow and peripheral blood cells after obtaining appropriate informed consent. Northern blot Total RNAs from control or treated cells were isolated using Tri Reagent Aliquots of RNA were electrophoresed and blotted onto nylon membranes, that hybridized to 32P-labelled probe. Western Blot Cells were lysed and. then were centrifugated. Protein concentration was determined by the Bradford assay.. Equivalent amounts of protein loaded and electrophoresed and were transferred to nitrocellulose membranes, that were incubated with the different primary antibodies:, Result and Discussion:. In the present study, we evaluated a pattern of different gene expression by Northern Blot and Western Blot analysis in bone marrow and peripheral blood cells from 16 CML patients at onset, from 2 patients in blast crisis evolved under IM treatment, and 14 imatinib-resistant patients. Some RNAs were underexpressed in most or all samples tested and never overexpressed (eg SIRT2, SIRT3, SIRT4 and SIRT5), while others were overexpressed in the great majority of samples and rarely, if ever, underexpressed (eg SIRT1, SIRT7, HSP90, iNOS)Furthermore, we examined the level of heat-shock related proteins HSP90 and bcl-2 in 2 patients during treatment with IM. and one patient IM-resistant by western Blot analysis: HSP90 and BCl-2 increased one patient during treatment with IM, while both protein levels was very high in one one patient IM-resistant These results suggest that the difference of genes expression might contribute to patterns of clinical response.

scholarly journals Tyrosine phosphorylation of CRKL in Philadelphia+ leukemia

Blood ◽  
1994 ◽  
Vol 84 (6) ◽  
pp. 1731-1736 ◽  
Author(s):  
J ten Hoeve ◽  
RB Arlinghaus ◽  
JQ Guo ◽  
N Heisterkamp ◽  
J Groffen
Keyword(s):  
Tyrosine Phosphorylation ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Catalytic Domain ◽  
Lymphoblastic Leukemia ◽  
Adaptor Protein ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells

Abstract The chimeric BCR/ABL protein is characteristic of Philadelphia (Ph)+ leukemia because it is the direct product of the Ph translocation and it has been shown to play a causal role in the genesis of leukemia. The BCR/ABL protein exhibits a deregulated tyrosine-kinase activity capable of phosphorylating different cellular substrates in vivo and in vitro. CRKL, an adaptor protein consisting of SH2 and SH3 domains in the absence of a catalytic domain, is one potential in vivo substrate of BCR/ABL. Previous experiments have shown that CRKL is phosphorylated on tyrosine in the chronic myelogenous leukemia (CML) cell line K562 and that CRKL is a substrate for ABL and for BCR/ABL in COS-1 cells. In the current study, we show that in peripheral blood cells a direct correlation exists between the presence of BCR/ABL and the phosphorylation status of CRKL. In Ph- peripheral blood cells, CRKL is present only in the nonphosphorylated form. In contrast, all BCR/ABL+ CML and acute lymphoblastic leukemia patient samples examined showed clear tyrosine-phosphorylation of CRKL. This result strongly suggests that CRKL is a biologically significant substrate for BCR/ABL and is likely to play a major role in the development of Ph+ leukemia.

scholarly journals Tyrosine phosphorylation of CRKL in Philadelphia+ leukemia

Blood ◽  
1994 ◽  
Vol 84 (6) ◽  
pp. 1731-1736 ◽  
Author(s):  
J ten Hoeve ◽  
RB Arlinghaus ◽  
JQ Guo ◽  
N Heisterkamp ◽  
J Groffen
Keyword(s):  
Tyrosine Phosphorylation ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Catalytic Domain ◽  
Lymphoblastic Leukemia ◽  
Adaptor Protein ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells

The chimeric BCR/ABL protein is characteristic of Philadelphia (Ph)+ leukemia because it is the direct product of the Ph translocation and it has been shown to play a causal role in the genesis of leukemia. The BCR/ABL protein exhibits a deregulated tyrosine-kinase activity capable of phosphorylating different cellular substrates in vivo and in vitro. CRKL, an adaptor protein consisting of SH2 and SH3 domains in the absence of a catalytic domain, is one potential in vivo substrate of BCR/ABL. Previous experiments have shown that CRKL is phosphorylated on tyrosine in the chronic myelogenous leukemia (CML) cell line K562 and that CRKL is a substrate for ABL and for BCR/ABL in COS-1 cells. In the current study, we show that in peripheral blood cells a direct correlation exists between the presence of BCR/ABL and the phosphorylation status of CRKL. In Ph- peripheral blood cells, CRKL is present only in the nonphosphorylated form. In contrast, all BCR/ABL+ CML and acute lymphoblastic leukemia patient samples examined showed clear tyrosine-phosphorylation of CRKL. This result strongly suggests that CRKL is a biologically significant substrate for BCR/ABL and is likely to play a major role in the development of Ph+ leukemia.

Aberrant CpG Island Methylation in AML Is Associated with Disease Progression and Relapse.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 564-564
Author(s):  
Heike Kroeger ◽  
Jaroslav Jelinek ◽  
Carlos E. Bueso-Ramos ◽  
Jean-Pierre J. Issa
Keyword(s):  
Dna Methylation ◽  
Bone Marrow ◽  
Disease Progression ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Cpg Islands ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells ◽  
Z Score ◽  
Methylation Index

The role of DNA methylation in relapse and progression of acute myelogenous leukemia (AML) is incompletely understood. We studied DNA methylation of 9 promoter-associated CpG islands of genes frequently hypermethylated in leukemic cell lines. These were NOR1, NPM2, HIN1, SLC26A4, CDH13, PGRA, PGRB, OLIG2 and the tumor suppressor gene p15INK4b. We examined bone marrow and/or peripheral blood cells collected at the time of diagnosis and at the first relapse from 32 patients (13 females, 19 males) with AML. The median age was 58 years (20–68), the median survival was 18 months (8–80), the median blast count was 64% (20–98), and 10 patients had additional solid tumors and/or lymphatic/hematologic malignancies. Bisulfite treatment of DNA, followed by PCR and pyrosequencing were used to quantitatively measure levels of cytosine methylation in promoter-associated CpG islands. We analyzed methylation data for individual genes and for a methylation index derived after Z-score (z = [value –mean]/standard deviation) transformation to equalize absolute differences between individual genes and we used paired t-tests for statistical analysis. Abnormal hypermethylation (≥10%) in bone marrow and peripheral blood cells at diagnosis was detected in all 9 investigated genes, with a range of 7/29 (24%) for HIN1 to 24/32 (75%) for CDH13. On average, an increase in methylation between diagnosis and relapse was detected in all genes, and was significant for CDH13 (mean 10%, p=0.0006), SLC26A4 (mean 7%, p=0.0012), HIN1 (mean 8%, p=0.0037), NPM2 (mean 7%, p=0.0073), p15INK4b (mean 13%, p=0.0081), NOR1 (mean 4%, p=0.0124), PGRB (mean 6%, p=0.0144), PGRA (mean 9%, p=0.0275), and OLIG2 (mean 3%, p=0.0732). When analyzed by change in methylation status: negative (methylation below 10%) turning positive (methylation ≥ 10%) and vice versa, of 238 analyses, 39 (16%) showed a negative to positive switch, 15 (6%) showed a positive to negative switch, and the remaining 184 (77%) were either positive or negative unchanged. Finally, when analyzed in individual patients, an increase in methylation was noted in 29 of 32 patients (91%). The median increase in methylation index between diagnosis and relapse calculated as a delta-Z-score was 30% (range from −10% to 147%), and was highly significant (p<0.0001). In summary, abnormal hypermethylation in bone marrow and/or peripheral blood cells from AML patients was detected in all investigated genes at diagnosis. Methylation levels further increased at relapse of the disease in 29 of 32 patients in 1 to 8 of 9 investigated genes. Based on quantitative analyses, we propose that methylation of CDH13, PGRB, PGRA and OLIG2 CpG islands are early markers for AML, while hypermethylation of HIN1, NPM2 and p15INK4b CpG islands is associated with disease progression and predominantly appears at relapse. Thus, aberrant hypermethylation is clearly associated with disease progression and relapse in AML, and likely mediates drug resistance in this setting. Increase of Methylation Index in Relapse Increase of Methylation Index in Relapse

Sign in / Sign up

Export Citation Format

Share Document