Determination of somatic mutant frequencies at glycophorin A and T-cell receptor loci for biodosimetry of prolonged irradiation.

PURPOSE: To determine the variant frequencies (VF) at glycophorin A (GPA) and T-cell receptor (TCR) loci in persons exposed to prolonged ionizing radiation at different doses and to assess the significance of the GPA and TCR assays for biodosimetry of prolonged irradiation. MATERIALS AND METHODS: The VF values were determined by means of flow cytometry in 120 persons exposed between 1968-1996. Most exposures were in Chernobyl clean-up workers in 1986-1987. RESULTS: A significant correlation was shown between the NO GPA variant cell frequency and dose (r = 0.61, p < 0.0001). The slope of the linear regression was 6.3 x 10(-6) NO mutant cells/Gy. Dose-dependent increase in the TCR VF was found in the group with recent exposures (slope 2.1 x 10(-4) variant cells/Gy, r = 0.75, p = 0.0002). In the Chernobyl clean up workers who received doses less than 0.25 Gy the TCR VF unlike the GPA VF was significantly higher than in the control non-irradiated individuals (p < 0.01 and p > 0.05 respectively). CONCLUSIONS: The GPA assay has limited potential to be used as a biodosimeter of prolonged irradiation, at least in dose interval up to 2.0 Gy. The TCR assay is likely to have greater potential in estimation of recent radiation exposure than the GPA assay.

Intracellular calcium signaling by Jurkat T-lymphocytes exposed to a 60 Hz magnetic field.

To explore possible biochemical mechanisms whereby electromagnetic fields of around 0.1 mT might affect immune cells or developing cancer cells, we studied intracellular calcium signaling in the model system Jurkat E6-1 human T-leukemia cells during and following exposure to a 60 Hz magnetic field. Cells were labeled with the intracellular calcium-sensitive fluorescent dye Fluo-3, stimulated with a monoclonal antibody against the cell surface structure CD3 (associated with ligand-stimulated T-cell activation), and analyzed on a FACScan flow-cytometer for increases in intensity of emissions in the range of 515-545 nm. Cells were exposed during or before calcium signal-stimulation to 0.15 mTrms 60 Hz magnetic field. The total DC magnetic field of 78.2 microT was aligned 17.5 degrees off the vertical axis. Experiments used both cells cultured at optimal conditions at 37 degrees C and cells grown under suboptimal conditions of 24 degrees C, lowered external calcium, or lowered anti-CD3 concentration. These experiments demonstrate that intracellular signaling in Jurkat E6-1 was not affected by a 60 Hz magnetic field when culture and calcium signal-stimulation were optimal or suboptimal. These results do not exclude field-induced calcium-related effects further down the calcium signaling pathway, such as on calmodulin or other calcium-sensitive enzymes.

Signal transduction in lymphocytes after low dose radiation.

Mice were given whole-body irradiation (WBI) with 75 mGy X-rays which had previously been found to stimulate immunologic functions. This low dose radiation (LDR) potentiated [Ca2+]i mobilization in thymic and splenic lymphocytes in response to Con A and anti-CD3 McAb and activated protein kinase C in T and B lymphocytes of the spleen. The expression of TCR/CD3 molecules on the thymocytes was enhanced after LDR indicating an expedited maturation and differentiation process in the thymus. The changes in TCR/CD3 expression and [Ca2+]i mobilization in response to McAb-CD3 after LDR was found to be highly correlated. Meanwhile the transcription of c-fos and c-jun genes was up-regulated beginning 3 hours after LDR. The expression of IL2R in active thymocytes was potentiated 24 hours after LDR which coincided with the previous finding of increased secretion of IL2 by splenocytes after WBI with 75 mGy X-rays. It is first reported in the present paper that LDR could stimulate immunologic functions through facilitation of the signal transduction process in the lymphocytes.