Abstract
An experimental model system is presented that allows the identification and follow-up of mice exposed to ionizing radiation using flow-cytometric measurements of peripheral blood cells. In an experiment, properties of peripheral blood cells were analysed with flow cytometry for a rapid identification of individuals exposed to radiation. Individuals were then followed longitudinally in an attempt to identify those developing neoplasias. Male CBA mice, 25 days old, were subjected to fractionated x-irradiation (4 × 1.31 Gy) to induce haematopoietic malignancies. By repeated blood sampling followed by flow cytometry, frequencies of micronucleated erythrocytes and of proliferating nucleated cells were determined. Neoplasias were diagnosed by histopathology. Five days after the end of radiation exposure, increased frequencies of proliferating cells, polychromatic erythrocytes and micronucleated normochromatic erythrocytes clearly distinguished the exposed group from the control group. Increased cell proliferation in peripheral blood cells could be used to identify animals with manifest tumours, although these animals were at a late stage of tumour development. Animals with thymic lymphoma (not generalized) could not be identified with the flow-cytometric parameters used. We consider that this model system has a potential use when a small number of risk individuals need to be identified and monitored within a large population.
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Abramsson-Zetterberg L, Grawé J, Zetterberg G (1995) Flow-cytometric analsis of micronucleus induction in mice by internal exposure to137Cs at very low dose rates. Int J Radiat Biol (in press)
Andreeff M (1990) Flow cytometry of leukemia. In: Melamed MR, Lindmo T, Mendelsohn ML (eds) Flow cytometry and sorting, 2nd edn. Wiley-Liss, New York, pp 697–724
Bonvier J, Humblet C, Rongy A-M, Delvenne C, Delvenne P, Greimers R, Thiry A, Courtoy R, Defresne MP (1990) Cellular events in radiation-induced lymphomagenesis. Int J Radiat Biol 57:693–698
Carlton JC, Terry NHA, White RA (1991) Measuring potential doubling times of murine tumours using flow cytometry. Cytometry 12:645–650
Crippen TL, Jones IM (1989) Cell proliferation in the bone marrow, thymus and spleen of mice studied by continuous, in vivo bromodeoxycytidine labelling and flow cytometric analysis. Cell Tissue Kinet 22:203–212
Cristaldi M, Ieradi LA, Mascanzoni D, Matte T (1991) Environmental impact of the Chernobyl accident: mutagenesis in bank voles from Sweden. Int J Radiat Biol 59:31–40
Cronkite EP, Bond VP, Carsten AL, Inoue T, Miller ME, Bullis JE (1987) Effects of low level radiation upon the hematopoietic stem cell: implications for leukemogenesis. Radiat Environ Biophys 26:103–114
Dolbeare F, Gratzner H, Pallavicini MG, Gray JW (1983) Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc Natl Acad Sci USA 80:5573–5577
Erp PEJ van, Brons PPT, Boezeman JBM, Jongh GJ de, Bauer FW (1988) A rapid flow cytometric method for bivariate bromodeoxyuridine/DNA analysis using simultaneous proteolytic enzyme digestin and acid denaturation. Cytometry 9:627–630
Giordano M, Riccardi A, Danova M, Brugnatelli S, Mazzini G (1991) Cell proliferation of human leukemia and solid tumors studied with in vivo bromodeoxyuridine and flow cytometry. Cancer Detect Prev 15:391–396
Grawé J, Zetterberg G, Amnéus H (1992) Flow-cytometric enumeration of micronucleated polychromatic erythrocytes in mouse peripheral blood. Cytometry 13:750–758
Grawé J, Zetterberg G, Amnéus H (1993) Effects of extended low dose-rate exposure to137Cs detected by flow cytometric enumeration of micronucleated erythrocytes in mouse peripheral blood. Int J Radiat Biol 63:339–347
Hollander Z, Loken MR (1988) Simultaneous analysis of DNA content and surface antigens in human bone marrow. Cytometry 9:485–490
Järplid B (1968) Radiation induced assymetry and lymphoma of thymus in mice. Acta Radiol Suppl (Stockh) 279:1
Jenssen D, Ramel C (1976) Dose response at low doses of X-irradiation and MMS on the induction of micronuclei in mouse erythroblasts. Mutat Res 41:311–320
Johannisson A, Amnéus H (1993) Characterization and enumeration of spontaneously proliferating leukocytes by multiparameter flow cytometry. Cell Prolif 26:317–326
Johannisson A, Eriksson B, Amnéus H, Zetterberg G (1992) Attempts to use the HPRT-assay as an automated short-term monitor for an acute exposure to mutagens. Cell Biol Toxicol 8:233–253
Kaplan HS, Brown MB (1952) A quantitative dose-response study of lymphoid tumor development in irradiation C57 black mice. J NCI 13:185
Mavournin KH, Blakey DH, Cimino MC, Salamone MF, Heddle JA (1990) The in vivo micronucleus assay in mammalian bone marrow and peripheral blood. A report of the U.S. Environmental protection agency gene-tox program. Mutat Res 239:29–80
Melamed MR, Lindmo T, Mendelsohn ML (eds) (1990) Flow cytometry and sorting, 2nd edn. Wiley-Liss, New York
Muto M, Kubo E, Kamisaku H, Sado T (1990) Phenotypic characterization of thymic prelymphoma cells of B 10 mice treated with split-dose irradiation. J Immunol 144:849–853
Pattengale PK, Frith CH (1983) Immunomorphological classification of spontaneous lymphoid cell neoplasms occurring in female BALB/c mice. J NCI 70:169–179
Riccardi A, Danova M, Wilson G, Ucci G, Dörmer P, Mazzini G, Drugnatelli S, Girino M, McNally NJ, Ascari E (1988) Cell kinetics in human malignancies studied with in vivo administration of bromodeoxyuridine and flow cytometry. Cancer Res 48:6238–6245
Rongy A-M, Defresne M-P, Greimers R, Humblet C, Boniver J (1987) Relations entre la capacité de repeuplement thymique et de formation de lymphomes exprimée par des cellules préleucémiques radio-induites chez la souris C57BL/Ka. C R Soc Biol (Paris) 181:88–93
Rongy A-M, Humblet C, Lelievre P, Greimers R, Defresne M-P, Boniver J (1990) Abnormal thymocyte subpopulations in split dose irradiated C57BL/Ka mice before the onset of lymphomas. Thymus 16:7–27
Shapiro HM (1988) Practical flow cytometry, 2nd edn. Liss, New York
Sprecher E, Giloh H, Rahamim E, Yefenof E, Becker Y (1989) Cytofluorometric analysis of thymic interdigitating cells from C57BL/6 mice prior and after leukemogenic X-irradiation. Leuk Res 9:799–809
Uma Devi P, Sharma ASKVS (1990) Mouse bone-marrow response to low doses of whole-body gamma irradiation: induction of micronuclei. Int J Radiat Biol 57:97–101
Young IT (1977) Proof without prejudice: use of the Kolmogorov-Smirnov test for analysis of histograms from flow systems and other sources. J Histochem Cytochem 25:935–941
Zetterberg G, Grawé J (1993) Flow-cytomeric analysis of micronucleus induction in mouse erythrocytes by γ-irradiation at very low dose rates. Int J Radiat Biol 65:555–564
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Johannisson, A., Grawé, J., Nilsson, A. et al. Flow-cytometric identification and follow-up of mice exposed to x-irradiation: Evaluation of a model system. Radiat Environ Biophys 34, 161–168 (1995). https://doi.org/10.1007/BF01211543
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DOI: https://doi.org/10.1007/BF01211543