Clinical Results on the Use of Human Ceruloplasmin in Aplastic Anemia

Shimizu, M.

doi:10.1046/j.1537-2995.1979.19680104101.x

Cited by 15 publications

(7 citation statements)

References 4 publications

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“…The studies with patients have not been confirmed in studies of mice with a disrupted Cp gene in which plasma hemoglobin levels were normal (6). Finally, a role of Cp in erythropoiesis has been shown directly in aplastic anemia patients; injection of purified Cp rapidly and dramatically increases both reticulocyte number and hemoglobin level (61). To our knowledge, Cp is the first positive acute phase reactant protein shown to be regulated by HIF-1 activation.…”

Section: Discussionmentioning

confidence: 97%

Role of Hypoxia-inducible Factor-1 in Transcriptional Activation of Ceruloplasmin by Iron Deficiency

Mukhopadhyay

Mazumder

Fox

2000

Journal of Biological Chemistry

258

163

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A role of the copper protein ceruloplasmin (Cp) in iron metabolism is suggested by its ferroxidase activity and by the tissue iron overload in hereditary Cp deficiency patients. In addition, plasma Cp increases markedly in several conditions of anemia, e.g. iron deficiency, hemorrhage, renal failure, sickle cell disease, pregnancy, and inflammation. However, little is known about the cellular and molecular mechanism(s) involved. We have reported that iron chelators increase Cp mRNA expression and protein synthesis in human hepatocarcinoma HepG2 cells. Furthermore, we have shown that the increase in Cp mRNA is due to increased rate of transcription. We here report the results of new studies designed to elucidate the molecular mechanism underlying transcriptional activation of Cp by iron deficiency. The 5-flanking region of the Cp gene was cloned from a human genomic library. A 4774-base pair segment of the Cp promoter/enhancer driving a luciferase reporter was transfected into HepG2 or Hep3B cells. Iron deficiency or hypoxia increased luciferase activity by 5-10-fold compared with untreated cells. Examination of the sequence showed three pairs of consensus hypoxia-responsive elements (HREs). Deletion and mutation analysis showed that a single HRE was necessary and sufficient for gene activation. The involvement of hypoxia-inducible factor-1 (HIF-1) was shown by gel-shift and supershift experiments that showed HIF-1␣ and HIF-1␤ binding to a radiolabeled oligonucleotide containing the Cp promoter HRE. Furthermore, iron deficiency (and hypoxia) did not activate Cp gene expression in Hepa c4 hepatoma cells deficient in HIF-1␤, as shown functionally by the inactivity of a transfected Cp promoter-luciferase construct and by the failure of HIF-1 to bind the Cp HRE in nuclear extracts from these cells. These results are consistent with in vivo findings that iron deficiency increases plasma Cp and provides a molecular mechanism that may help to understand these observations.

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Section: Discussionmentioning

confidence: 97%

Role of Hypoxia-inducible Factor-1 in Transcriptional Activation of Ceruloplasmin by Iron Deficiency

Mukhopadhyay

Mazumder

Fox

2000

Journal of Biological Chemistry

258

163

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“…The detailed procedure was described in the literature using o-dianisidine dihydrochloride (4,4'-diamino-3,3'-dimethoxybiphenyl) as a substrate. It was reported that human Cp treatment had beneficial effects on aplastic anemia patients by intravenous injection (Shimizu, 1979). However, the isolation and purification procedures of Cp are too lengthy and time-consuming, causing undesirable proteolytic degradation products.…”

Section: Resultsmentioning

confidence: 99%

A novel and one-step purification of human ceruloplasmin by acharan sulfate affinity chromatography

et al. 2009

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Human ceruloplasmin, a copper binding alpha(2)-glycoprotein, was purified by a single-step procedure using acharan sulfate affinity chromatography. Acharan sulfate was immobilized to amine-functionalized agarose matrix through carboxylic acids. Ceruloplasmin in human plasma was obtained from 0.4 M NaCl salt elution and characterized by SDS-PAGE (132 and 125 kDa), isoelectric focusing (pI 4.6), Western blotting, and MALDI-TOF-MS peptide mass fingerprinting. Ceruloplasmin was purified 106 fold with a specific oxidase activity of 0.53 U/mg protein.

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“…Recently, we have shown that ceruloplasmin (CP), the main serum Cu‐binding protein, is, by itself, a strong inducer of HL‐60 cell differentiation (Peled et al , 1998). In this context, it is interesting to note that Cp has been shown to have a therapeutic effect in patients with aplastic anaemia (Shimizu, 1979).…”

Section: Discussionmentioning

confidence: 99%

Cellular copper content modulates differentiation and self‐renewal in cultures of cord blood‐derived CD34⁺ cells

et al. 2002

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Summary. Several clinical observations have suggested that copper (Cu) plays a role in regulating haematopoietic progenitor cell (HPC) development. To further study this role we used an ex vivo system. Cord blood-derived CD34 + cells were cultured in liquid medium supplemented with Kitligand, FLt3, interleukin 6 (IL-6), thrombopoietin and IL-3. Under these conditions, Cu content, measured by atomic absorption, was 7 ng/10 7 cells. Modulation of intracellular Cu was achieved by supplementing the cultures with the Cu chelator tetraethylenepentamine, which reduced cellular Cu (4 ng/10 7 cells), or ceruloplasmin or Cu sulphate that elevated cellular Cu (18 and 14 ng/10 7 cells respectively). The results indicated that low Cu content delayed differentiation, as measured by the surface antigens CD34, CD14 and CD15, colony-forming unit (CFU) frequency and cell morphology, while high Cu accelerated differentiation compared with Cu unmanipulated cultures. As a result, expansion of total cells, CFU and CD34 + cells in low Cu was extended (12-16 weeks), and in high Cu was shortened (2-4 weeks), compared with control cultures (6-8 weeks). These effects required modulation of intracellular Cu only during the first 1-3 weeks of the culture; the long-term effects persisted thereafter, suggesting that the decision process for either self-renewal or differentiation is taken early during the culture. This novel method of controlling cell proliferation and differentiation by copper and copper chelators might be utilized for ex vivo manipulation of HPC for various clinical applications.

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Clinical Results on the Use of Human Ceruloplasmin in Aplastic Anemia

Cited by 15 publications

References 4 publications

Role of Hypoxia-inducible Factor-1 in Transcriptional Activation of Ceruloplasmin by Iron Deficiency

Role of Hypoxia-inducible Factor-1 in Transcriptional Activation of Ceruloplasmin by Iron Deficiency

A novel and one-step purification of human ceruloplasmin by acharan sulfate affinity chromatography

Cellular copper content modulates differentiation and self‐renewal in cultures of cord blood‐derived CD34⁺ cells

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Clinical Results on the Use of Human Ceruloplasmin in Aplastic Anemia

Cited by 15 publications

References 4 publications

Role of Hypoxia-inducible Factor-1 in Transcriptional Activation of Ceruloplasmin by Iron Deficiency

Role of Hypoxia-inducible Factor-1 in Transcriptional Activation of Ceruloplasmin by Iron Deficiency

A novel and one-step purification of human ceruloplasmin by acharan sulfate affinity chromatography

Cellular copper content modulates differentiation and self‐renewal in cultures of cord blood‐derived CD34+ cells

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Cellular copper content modulates differentiation and self‐renewal in cultures of cord blood‐derived CD34⁺ cells