umu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Erythrocyte Flow Cytometric Analysis in Congenital Dyserythropoietic Anemia Type III-Evaluation of Eosin-5´-Maleimide, CD55, and CD59
Umeå University, Faculty of Medicine, Department of Radiation Sciences, Oncology.
Umeå University, Faculty of Medicine, Department of Medical Biosciences.
Umeå University, Faculty of Medicine, Department of Medical Biosciences, Medical and Clinical Genetics.
Umeå University, Faculty of Medicine, Department of Public Health and Clinical Medicine, Family Medicine.
Show others and affiliations
2013 (English)In: Journal of Blood Disorders & Transfusion, ISSN 2155-9864, Vol. 121, no 23, 4791-4799 p.Article in journal (Refereed) Published
Abstract [en]

Introduction: Flow cytometry with eosin-5´-maleimide (EMA), anti-CD55 and anti-CD59 is commonly used when investigating non-autoimmune hemolytic anemias. Reduced fluorescence of EMA, typically detected in hereditary spherocytosis is also seen in congenital dyserythropoietic anemia type II (CDA II). Reduction of CD55 and CD59 characterizes paroxysmal nocturnal hemoglobinuria (PNH). We studied the flow cytometric profile of EMA, CD55 and CD59 on erythrocytes in congenital dyserythropoietic anemia type III (CDA III). 

Methods: Erythrocytes from 16 CDA III positive individuals, 14 CDA III negative relatives and three normal controls per assay were studied with flow cytometry after EMA staining. Flow cytometry after anti-CD55 and anti- CD59 was performed on erythrocytes from 12 CDA III positive and 7 CDA III negative relatives with one normal control per assay. 

Results: CDA III - erythrocytes exhibited marginally stronger fluorescence after EMA-staining than normal controls. Correlation between EMA fluorescence and erythrocyte volume was confirmed. CDA III subjects did not differ from normal controls concerning CD55 and CD59. 

Conclusion: The results of the present study indicate no abnormality of the erythrocyte membrane in CDA III and show that standard flow cytometry cannot be used to discriminate between CDA III and normal controls. 

Place, publisher, year, edition, pages
OMICS International , 2013. Vol. 121, no 23, 4791-4799 p.
National Category
Hematology
Identifiers
URN: urn:nbn:se:umu:diva-117449DOI: 10.4172/2155-9864.1000172OAI: oai:DiVA.org:umu-117449DiVA: diva2:907773
Available from: 2016-02-29 Created: 2016-02-29 Last updated: 2016-04-15Bibliographically approved
In thesis
1. Congenital Dyserythropoietic Anemia type III (CDA III): diagnostics, genetics and morbidity
Open this publication in new window or tab >>Congenital Dyserythropoietic Anemia type III (CDA III): diagnostics, genetics and morbidity
2016 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The Congenital Dyserythropoietic Anemias (CDA) are rare hereditary hemolytic disorders with large bi- to multi-nucleated erythroblasts in the bone marrow. Hemolysis is negative in a direct antiglobulin test (DAT). Based on morphology and clinical picture, three major forms of CDAs, type I, II, and III have been defined. CDA III, dominantly inherited, constitutes the rarest type with a majority of cases belonging to a family in Västerbotten, Sweden. The genetic background of CDA I and CDA II has been linked to mutations in CDAN1 and SEC23B respectively. The mutation of CDA III has been linked to 15q22 in earlier studies.

In this project we have defined the causative genetic lesion in two families with CDA III. The novel mutation KIF23 c.2747C>G (p.P916R) was shown to segregate with CDA III in the Swedish and American CDA III families and was absent in 356 healthy controls. KIF23 encodes mitotic kinesin-like protein 1 (MKLP1), which plays a central role in the last step of cytokinesis. RNAi-based knock-down and rescue experiments demonstrated that the p.P916R mutation causes cytokinesis failure in HeLa cells, resulting in increasing number of bi-nuclear cells, consistent with appearance of large multinucleated erythroblasts in CDA III patients. We conclude that CDA III is caused by a mutation in KIF23, encoding MKLP1, a conserved mitotic kinesin crucial for cytokinesis.

Flow cytometry with eosin-5´-maleimide (EMA), anti-CD55 and anti-CD59 is commonly used when investigating non-autoimmune hemolytic anemias. Reduced fluorescence of EMA, typically detected in hereditary spherocytosis, is also seen in CDA II, while reduction of CD55 and CD59 characterizes paroxysmal nocturnal hemoglobinuria (PNH). We studied the flow cytometric profile of EMA, CD55, and CD59 on erythrocytes in CDA III. We found no abnormality of the erythrocyte membrane in CDA III and concluded that standard flow cytometry cannot be used to discriminate between CDA III and normal controls.

In CDA I and CDA II a majority of patients, including those who are not transfusion dependent, suffer from iron overload, which, according to earlier studies, is not the case in CDA III. We found that individuals of the Västerbotten CDA III family carry mutations in the hemochromatosis (HFE) gene. Three CDA III patients with heterozygous or compound HFE mutations need treatment with phlebotomy due to iron overload. One of them carries heterozygous H63D mutation, which is not reported to lead to iron overload by itself in otherwise healthy individuals. We propose that molecular genetic testing of the HFE gene is indicated in all patients with CDA, including CDA III.

Place, publisher, year, edition, pages
Umeå: print och media, 2016. 54 p.
Series
Umeå University medical dissertations, ISSN 0346-6612 ; 1784
Keyword
congenital dyserythropoietic anemia, KIF23, hereditary hemochromatosis, iron overload, flow cytometry
National Category
Hematology
Identifiers
urn:nbn:se:umu:diva-117454 (URN)978-91-7601-424-0 (ISBN)
Public defence
2016-04-22, E04, By 6E, 901 85, Norrlands Universitets sjukhus, Umeå, 19:35 (Swedish)
Opponent
Supervisors
Available from: 2016-04-15 Created: 2016-02-29 Last updated: 2016-04-22Bibliographically approved

Open Access in DiVA

fulltext(655 kB)76 downloads
File information
File name FULLTEXT01.pdfFile size 655 kBChecksum SHA-512
db69df41b6d2c563d4d5a1203e545c1f8e333b9154f82071120295aa29fd4276d3e3c0b3f62a45fb71fe5c48b68a9433a72e2d673e095f45dd7825b15e3a2799
Type fulltextMimetype application/pdf

Other links

Publisher's full text

Search in DiVA

By author/editor
Liljeholm, MariaGrönlund, ElisabethGolovleva, IrinaSandström, HerbertWahlin, Anders
By organisation
OncologyDepartment of Medical BiosciencesMedical and Clinical GeneticsFamily Medicine
Hematology

Search outside of DiVA

GoogleGoogle Scholar
Total: 76 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 105 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf