Detection of the 4977 bp deletion of mitochondrial DNA in different human blood cells

Mohamed, S. A., Wesch, D., Blumenthal, A., Bruse, P., Windler, K., Ernst, M., Kabelitz, D., Oehmichen, M. and Meissner, C. (2004) Detection of the 4977 bp deletion of mitochondrial DNA in different human blood cells. Experimental Gerontology, 39 2: 181-188. doi:10.1016/j.exger.2003.10.011


Author Mohamed, S. A.
Wesch, D.
Blumenthal, A.
Bruse, P.
Windler, K.
Ernst, M.
Kabelitz, D.
Oehmichen, M.
Meissner, C.
Title Detection of the 4977 bp deletion of mitochondrial DNA in different human blood cells
Journal name Experimental Gerontology   Check publisher's open access policy
ISSN 0531-5565
1873-6815
Publication date 2004-02
Sub-type Article (original research)
DOI 10.1016/j.exger.2003.10.011
Volume 39
Issue 2
Start page 181
End page 188
Total pages 8
Place of publication Philadelphia, PA, United States
Publisher Elsevier
Language eng
Formatted abstract
As recently reported, it is possible to detect and quantify the amount of the deleted human mitochondrial DNA (mtDNA) in whole blood, platelets and peripheral blood mononuclear cells using real-time PCR. The aim of this study was to identify the cell types in human blood carrying the 4977 bp deleted mtDNA and their accumulation with regard to donor age. Whole blood from 10 healthy donors (five individuals aged from 19 to 22 years, five aged from 57 to 61 years) was separated in various cell populations such as granulocytes, B cells/monocytes and T cells. Purity of the cell isolates was determined by flow cytometry. Total DNA was extracted and 250 ng DNA of each cell type was subjected to PCR using fluorescent-labelled primer pairs. The specific PCR product of the 4977 bp deletion was quantified using an automated detection system. The accumulation of the 4977 bp deletion was more pronounced in T lymphocytes and granulocytes in comparison to B lymphocytes/monocytes. The amount of the 4977 bp deletion in whole blood varied from 0 to 0.00018%, in T lymphocytes from 0.00009 to 0.00160%, in granulocytes from 0 to 0.00162% and in the B lymphocyte/monocyte fraction from 0 to 0.00025%. The higher amount of the deletion in T lymphocytes may be due to a subset of lymphocytes with a longer lifespan thus facilitating the accumulation of mitochondrial damage. The higher amount in granulocytes could have the explanation in the higher release of free radicals for prevention of infectious diseases, because free radicals are supposed to damage the macromolecules of this cell type. The 10 donors displayed differences in the pattern of the accumulation with regard to the different cell types, but no age-dependent accumulation was observed. Differences of the accumulation pattern may be due to actual individual living behaviour or environmental factors.
Keyword Human peripheral blood cells
Mitochondrial DNA
Fluorescent-labelled primer
4977 bp deletion
Aging
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: UQ Diamantina Institute Publications
 
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Created: Thu, 10 Mar 2011, 10:01:54 EST