10.6084/M9.FIGSHARE.13624046.V1
Patrick Ostheim
Patrick
Ostheim
University of Ulm
Alan Don Mallawaratchy
Alan Don
Mallawaratchy
Thomas Müller
Thomas
Müller
University Hospital Regensburg
Friedrich-Loeffler-Institut
University of Bern
Simone Schüle
Simone
Schüle
University of Ulm
Cornelius Hermann
Cornelius
Hermann
University of Ulm
Tanja Popp
Tanja
Popp
Stefan Eder
Stefan
Eder
University of Ulm
Stephanie E. Combs
Stephanie E.
Combs
Helmholtz Zentrum München
Technical University of Munich
German Cancer Research Center
Matthias Port
Matthias
Port
University of Ulm
Michael Abend
Michael
Abend
University of Ulm
Acute Radiation Syndrome-related gene expression in irradiated peripheral blood cell populations
<p>In a nuclear or radiological event, an early diagnostic or prognostic tool is needed to distinguish the worried well from low-exposed and those individuals who may later develop life-threatening hematologic acute radiation syndrome (H-ARS). In previous studies, we identified and validated genes in peripheral blood for this purpose. To gain a deeper understanding and to make methodological improvements, we examined the contribution of the peripheral blood´s cell populations on radiation-induced gene expression changes.</p> <p>EDTA whole blood from six healthy donors was X-irradiated with 0 and 4 Gy, cultured <i>in vitro</i> for 24 h and cell populations of T-lymphocytes (CD3), B-lymphocytes (CD19), NK-cells (CD56) and granulocytes (CD15) were separated using immunomagnetic methods. Whole blood was used as a positive control to ensure the expected radiation-induced gene expression response based on previous examinations. Purity of cell separation and cell counts was validated using immunofluorescence imaging flow cytometry. After RNA isolation, gene expressions of <i>FDXR, DDB2, POU2AF1</i> and <i>WNT3</i> were examined in the cell populations and whole blood.</p> <p>The cell populations, on average, contributed to the total RNA amount with a ratio of 11.6 for T-lymphocytes: 1.2 for B-cells: 1.2 for NK-cells: 1.0 for granulocytes. In order to estimate the contribution of gene expression per cell population, the baseline (0 Gy) as well as the radiation-induced fold-change in gene expression relative to unexposed was considered for each gene. After considering all three parameters, the T-lymphocytes (74.8%/80.5%) contributed predominantly to the radiation-induced up-regulation observed for <i>FDXR/DDB2</i> and the B-lymphocytes (97.1%/83.8%) for down-regulated <i>POU2AF1/WNT3</i> with a similar effect on whole blood gene expression measurements reflecting a corresponding order of magnitude.</p> <p>T-lymphocytes and B-lymphocytes contributed predominantly to the radiation-induced up-regulation of <i>FDXR/DDB2</i> and down-regulation of <i>POU2AF1/WNT3</i>. Further separation of cell populations will not increase the diagnostic sensitivity, but complicate an efficient workflow. Also, this study identifies undesired limitations of widely used whole blood in vitro models, but it still underlines the use of <i>FDXR</i> and <i>DDB2</i> for biodosimetry purposes and <i>POU2AF1</i> and <i>WNT3</i> for effect prediction of acute health effects.</p>
Cell Biology
Genetics
Molecular Biology
59999 Environmental Sciences not elsewhere classified
Immunology
69999 Biological Sciences not elsewhere classified
Developmental Biology
Cancer
Hematology
Taylor & Francis
2021
2021-01-21
2023-01-06
Dataset
4454987 Bytes
10.1080/09553002.2021.1876953
10.6084/m9.figshare.13624046
CC BY 4.0