Kyoizumi S, Nakamura N, Hakoda M, Awa A A, Bean M A, Jensen R H, Akiyama M
Department of Radiobiology, Radiation Effects Research Foundation, Hiroshima, Japan.
Cancer Res. 1989 Feb 1;49(3):581-8.
A modified method was developed for measuring the frequency of variant erythrocytes at the glycophorin A locus using a single beam cell sorter (SBS). Fluorescein- or phycoerythrin-labeled monoclonal antibodies specific for the M or N glycophorin A alleles were used for the SBS assay. To prevent contamination of nucleated cells in the sorting windows, the nucleated cells in the fixed erythrocyte sample were stained with propidium iodide before flow sorting. Blood samples were obtained from atomic bomb survivors who were heterozygous for the MN blood type, and the frequencies of the hemizygous and homozygous variant of the M or N glycophorin A allele were measured by the SBS. For the three types of variants, hemizygotes for M and N allele (Nø and Mø) and homozygotes for M allele (MM), the variant frequency measured by the SBS correlated well with that previously determined by a dual beam cell sorter. Variant frequencies of the Nø, Mø, and MM cell types in atomic bomb survivors determined by SBS measurements were found to increase with radiation dose (DS86, kerma) as well as with the frequency of chromosome aberrations in lymphocytes.
开发了一种改进方法，用于使用单光束细胞分选仪（SBS）测量血型糖蛋白A位点变异红细胞的频率。将针对M或N血型糖蛋白A等位基因的荧光素或藻红蛋白标记的单克隆抗体用于SBS检测。为防止分选窗口中有核细胞的污染，在流式分选前，用碘化丙啶对固定红细胞样本中的有核细胞进行染色。从MN血型杂合的原子弹幸存者中采集血样，并用SBS测量M或N血型糖蛋白A等位基因的半合子和纯合子变异频率。对于三种类型的变异体，即M和N等位基因的半合子（Nø和Mø）以及M等位基因的纯合子（MM），SBS测量的变异频率与先前用双光束细胞分选仪测定的结果高度相关。通过SBS测量确定的原子弹幸存者中Nø、Mø和MM细胞类型的变异频率随辐射剂量（DS86，比释动能）以及淋巴细胞中染色体畸变频率的增加而增加。
