Quantification of γ-H2AX Foci in Human Lymphocytes: A Method for Biological Dosimetry after Ionizing Radiation Exposure

Abstract Recent studies have suggested that visualization of γ-H2AX nuclear foci can be used to estimate exposure to very low doses of ionizing radiation. Although this approach is widely used for various purposes, its suitability for individual human biodosimetry has not yet been assessed. We therefore conducted such an assessment with the help of available software for observing and automatically scoring γ-H2AX foci. The presence of γ-H2AX foci was evaluated in human peripheral blood lymphocytes exposed ex vivo to γ rays in a dose range of 0.02 to 2 Gy. We analyzed the response of γ-H2AX to ionizing radiation in relation to dose, time after exposure, and individual variability. We constructed dose–effect calibration curves at 0.5, 8 and 16 h after exposure and evaluated the threshold of detection of the technique. The results show the promise of automatic γ-H2AX scoring for a reliable assessment of radiation doses in a dose range of 0.6 Gy to 2 Gy up to 16 h after exposure. This γ-H2AX-based assay may be useful for biodosimetry, especially for triage to distinguish promptly among individuals the ones who have received negligible doses from those with significantly exposures who are in need of immediate medical attention. However, additional in vivo experiments are needed for validation.

[1]  P. R. Barber,et al.  Gamma-H 2 AX Foci Counting : Image processing and control software for high-content screening , 2007 .

[2]  A. Takahashi,et al.  Heat induces gammaH2AX foci formation in mammalian cells. , 2008, Mutation research.

[3]  George Iliakis,et al.  Computational Methods for Analysis of Foci: Validation for Radiation-Induced γ-H2AX Foci in Human Cells , 2006, Radiation research.

[4]  Ruth C Wilkins,et al.  Canadian Cytogenetic Emergency Network (CEN) for biological dosimetry following radiological/nuclear accidents , 2007, International journal of radiation biology.

[5]  C. Streffer,et al.  Democratic Republic of the Congo , 2009, Prevention of Tax Treaty Abuse – Third Peer Review Report on Treaty Shopping.

[6]  Vicky Goh,et al.  Leukocyte DNA damage after multi-detector row CT: a quantitative biomarker of low-level radiation exposure. , 2007, Radiology.

[7]  E. Rogakou,et al.  Megabase Chromatin Domains Involved in DNA Double-Strand Breaks in Vivo , 1999, The Journal of cell biology.

[8]  E. Rogakou,et al.  DNA Double-stranded Breaks Induce Histone H2AX Phosphorylation on Serine 139* , 1998, The Journal of Biological Chemistry.

[9]  F. Cucinotta,et al.  Induction and quantification of gamma-H2AX foci following low and high LET-irradiation. , 2006, International journal of radiation biology.

[10]  D. Lloyd,et al.  Criticality accident dosimetry by chromosomal analysis. , 2004, Radiation protection dosimetry.

[11]  P. Olive,et al.  Phosphorylation of histone H2AX as a measure of radiosensitivity. , 2004, International journal of radiation oncology, biology, physics.

[12]  Takeo Ohnishi,et al.  Does γH2AX foci formation depend on the presence of DNA double strand breaks , 2005 .

[13]  V. Durand,et al.  The cytogenetic dosimetry of recent accidental overexposure. , 2001, Cellular and molecular biology.

[14]  J. Savage Radiation-induced chromosomal aberrations in the plant Tradescantia: Dose-response curves I. Preliminary considerations , 1975 .

[15]  Michael Uder,et al.  In vivo formation and repair of DNA double-strand breaks after computed tomography examinations. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[16]  Takeo Ohnishi,et al.  Does gammaH2AX foci formation depend on the presence of DNA double strand breaks? , 2005, Cancer letters.

[17]  M. Engelhard,et al.  gamma-H2AX foci formation in peripheral blood lymphocytes of tumor patients after local radiotherapy to different sites of the body: Dependence on the dose-distribution, irradiated site and time from start of treatment , 2007, International journal of radiation biology.

[18]  Kai Rothkamm,et al.  Evidence for a lack of DNA double-strand break repair in human cells exposed to very low x-ray doses , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Barry S Rosenstein,et al.  Development of an Automated γ-H2AX Immunocytochemistry Assay , 2009, Radiation research.

[20]  Borivoj Vojnovic,et al.  Gamma-H2AX foci counting: image processing and control software for high-content screening , 2007, SPIE BiOS.

[21]  Philippe Voisin,et al.  Why can't we find a better biological indicator of dose? , 2004, Radiation protection dosimetry.

[22]  Jan Nyman,et al.  DNA double strand break quantification in skin biopsies. , 2004, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[23]  G. Cuttone,et al.  Induction and Repair of DNA Double-Strand Breaks in Human Cells: Dephosphorylation of Histone H2AX and its Inhibition by Calyculin A , 2005, Radiation research.

[24]  J. Harper,et al.  Induction and quantification of γ-H2AX foci following low and high LET-irradiation , 2006 .

[25]  Matthew A. Coleman,et al.  Candidate protein biodosimeters of human exposure to ionizing radiation , 2006, International journal of radiation biology.