Technical Report No. 3-91
Development of a flow-cytometric HLA-A locus mutation assay for human peripheral blood lymphocytes
Kushiro J, Hirai Y, Kusunoki Y, Kyoizumi S, Kodama Y, Wakisaka A, Jeffreys AJ, Cologne JB, Nakamura N, Akiyama MEditor’s note: A publication based on this report was published in Mutat Res 272:17-29, 1992.
Summary
A flow-cytometric technique was developed to measure the frequency of mutant lymphocytes lacking expression of HLA-A2 or A24 allele products among donors heterozygous for HLA-A2 or A24.
It was found that the mutant frequency of lymphocytes in peripheral blood was on 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 blot analysis. A small-scale study on atomic bomb survivors did not show a significant dose effect.
An in vitro mutagenesis study showed that the mutant frequency at the HLA-A24 locus increased at a rate of roughly 2 x 10-4/Gy, about 10 times greater than that reported at the X-chromosomal hypoxanthine phosphoribosyltransferase locus in lymphocytes. These mutants were found to be mostly derived from large chromosomal deletions.
A flow-cytometric technique was developed to measure the frequency of mutant lymphocytes lacking expression of HLA-A2 or A24 allele products among donors heterozygous for HLA-A2 or A24.
It was found that the mutant frequency of lymphocytes in peripheral blood was on 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 blot analysis. A small-scale study on atomic bomb survivors did not show a significant dose effect.
An in vitro mutagenesis study showed that the mutant frequency at the HLA-A24 locus increased at a rate of roughly 2 x 10-4/Gy, about 10 times greater than that reported at the X-chromosomal hypoxanthine phosphoribosyltransferase locus in lymphocytes. These mutants were found to be mostly derived from large chromosomal deletions.