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CONTRIBUTION DE LA CYTOFLUORIMETRIE EN FLUX A L'EVALUATION DE LA SUREXPOSITION AUX RADIATIONS IONISANTES

Objective


Flow cytofluorimetry is used for the assessment of over exposure to ionising radiation. Work concentrated mainly on the labelling of chromosomal kinetochores. Different techniques were used in order to homogeneously stain chromosomes and to reduce background due to nonspecific staining by fluorescent antibodies. Staining and detection of reticulocytes was performed using thiazole orange.
THE PROPOSAL INVOLVES 3 ASPECTS:
1. THE UTILISATION OF RADIATION-INDUCED CHROMOSOME ABERRATIONS.
THE METHODS OF ANALYSING CHROMOSOME ABERRATIONS BY FLOW CYTOMETRY WILL BE APPLIED TO HUMAN LYMPHOCYTES IRRADIATED IN VITRO IN 1985. THE DOSE EFFECT RELATIONSHIPS WILL BE DETERMINED AND A COMPUTER PROGRAMME WILL BE DEVELOPED TO REFINE THE ANALYSIS IN 1985/86.
AN IN VIVO STUDY IN PRIMATES WILL BE CARRIED OUT IN PARALLEL AND THE CHOICE OF THE PRIMATE TO BE USED WILL BE MADE IN 1985.
2. ANALYSIS OF CHROMOSOME ABERRATIONS BY MEANS OF MONOCLONAL ANTIBODIES.
IN 1985 THE PROBLEM WILL BE APPROACHED BY STUDYING WHETHER ONE CAN DETECT THE CENTROMER WITH AN ANTI-CENTROMERE SERUM. THIS HAS BEEN PREVIOUSLY SHOWN IN FIXED CELLS AND WILL NOW (1985/86) BE EXTENDED TO CHROMOSOMES IN SUSPENSION. FOR THIS PURPOSE, A METHOD TO AMPLIFY FLUORESCENCE WILL BE DEVELOPED.
FROM 1986 THE USEFULNESS OF THIS MARKER FOR THE IDENTIFICATION OF DICENTRIC CHROMOSOMES AND ACENTRIC FRAGMENTS WILL BE INVESTIGATED BY COMPARING DIFFERENT FLUORESCENCE SIGNALS (DNA AND CENTROMERE). THE ANALYSIS OF THE RESULTS BY MEANS OF A SPECIAL COMPUTER PROGRAMME WILL BE CARRIED OUT IN 1986/87.
FROM 1987 THE DOSE EFFECT RELATIONS WILL BE VALIDATED BY COMPARING THEM WITH THOSE OBTAINED BY CONVENTIONAL DETERMINATION OF THE CARYOTYPE.
3. STUDY OF RADIATION-INDUCED CHANGES IN THE MEMBRANE.
KUBASOVA AND COWORKERS HAVE FOUND CHANGES IN THE SURFACE OF BLOOD CELLS BY MEANS OF RADIOACTIVE LECTINES AFTER IRRADIATION WITH DOSES OF MORE THAN 0.25 GY.
THE PROPOSED STUDY INTENDS TO DETECT THESE CHANGES IN THE MEMBRANE WITH THE HELP OF FLOW CYTOMETRY. THE STUDY SHALL PROCEED AS FOLLOWS: CHARACTERISATION OF THE SUBPOPULATIONS OF THE BLOOD CELLS, FIXATION OF THE LECTINES, DETERMINATION OF THE SMALLEST CHANGES IN FLUORESCENT MOLECULES DETECTABLE BY FLOW CYTOMETRY.
THE STUDY WILL BE EXTENDED TO OTHER SURFACE ANTIGENS WHICH MAY BE MARKED WITH A FLUOROCHROME AND WILL BE CARRIED OUT ACCORDING TO THE FOLLOWING SCHEDULE:
1985-1986 FEASIBILITY STUDY: 1987-1988 VALIDATION OF THE METHOD FOR DETERMINING DOSE EFFECT RELATIONSHIPS: 1987-1989 APPLICATION OF THE METHOD FOR THE EVALUATION OF PROTECTED AREAS (HETEROGENEITY OF ABSORBED DOSES).

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

Commissariat à l'Energie Atomique (CEA)
Address
Departement De Protection Sanitaire
92260 Fontenay-aux-roses
France