Development of a flow-cytometric HLA-A locus mutation assay for human peripheral blood lymphocytes. |
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Authors: | J Kushiro Y Hirai Y Kusunoki S Kyoizumi Y Kodama A Wakisaka A Jeffreys J B Cologne K Dohi N Nakamura |
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Affiliation: | Department of Radiobiology, Radiation Effects Research Foundation, Hiroshima, Japan. |
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Abstract: | A flow-cytometric technique was developed to measure the frequency of variant lymphocytes lacking expression of HLA-A2 or A24 allele products among donors heterozygous for HLA-A2 or A24. It was found that the variant frequency of lymphocytes in peripheral blood was of the order of 10(-4) and increased with donor age. Molecular analyses of mutant clones revealed that about one-third were derived from somatic recombinations and that the remaining two-thirds did not show any alterations after Southern blotting analysis. In contrast, mutants obtained after in vitro X-ray mutagenesis study were found to be mostly derived from large chromosomal deletions. A small-scale study on atomic bomb survivors did not show a significant dose effect. |
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