Human blood PIG-A mutation and micronucleated reticulocyte flow cytometric assays: Method optimization and evaluation of intra- and inter-subject variation.
Human blood PIG-A mutation and micronucleated reticulocyte flow cytometric assays: Method optimization and evaluation of intra- and inter-subject variation.
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DOI：
10.1002/em.22393
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发表时间：
2020-10
影响因子：
2.8
通讯作者：
Dertinger SD
中科院分区：
文献类型：
--
作者：
Torous DK;Avlasevich SL;Khattab MG;Baig A;Saubermann LJ;Chen Y;Bemis JC;Lovell DP;Walker VE;MacGregor JT;Dertinger SD
我们之前介绍了基于流式细胞术的方法来评估啮齿动物和人类血液样本中微核网织红细胞(MN-RET)和PIG-A突变表型网织红细胞(Mut RET)的发生率。本报告描述了人类血液分析的重要方法改进，包括在MN-RET评分之前对CD71阳性网织红细胞进行免疫磁性浓缩，以及为以后的PIG-A分析储存冷冻血液的程序。量化了基于14名受试者血液样本的MN-RET和MUT RET频率的技术重复变异性，基于6名受试者连续采血的受试者内变异性，以及基于多达344名0至73岁受试者的受试者间变异性。受试者之间的差异解释了观察到的两个终点的大部分变异性(≥77%)，受试者内和技术上的重复变异性要低得多。在剔除两个极端异常值后，MN-RET(0.15±0.10%)和MUT RET(4.7±5.0/百万)的平均值和标准差明显存在较大程度的受试者间差异。研究了年龄和性别对受试者间变异的影响，发现两种因素均不影响MN-RET，但都影响MT-RET频率。11岁的受试者MUT RET值最低，男性的MUT RET频率略高于女性。这些结果表明，MN-RET和MUT RET是自动兼容的遗传毒性生物标记物，将毒理学感兴趣的物种与包括人类种群联系起来。这些数据将有助于正确设计未来的人类研究，包括这些遗传毒性的生物标记物，并突出了旨在确定本文报告的个体间变异的来源的额外工作的必要性。
We previously described flow cytometry-based methods for scoring the incidence of micronucleated reticulocytes (MN-RET) and PIG-A mutant phenotype reticulocytes (MUT RET) in rodent and human blood samples. The current report describes important methodological improvements for human blood analyses, including immunomagnetic enrichment of CD71-positive reticulocytes prior to MN-RET scoring, and procedures for storing frozen blood for later PIG-A analysis. Technical replicate variability in MN-RET and MUT RET frequencies based on blood specimens from 14 subjects, intra-subject variability based on serial blood draws from 6 subjects, and inter-subject variation based on up to 344 subjects age 0 to 73 years were quantified. Inter-subject variation explained most of the variability observed for both endpoints (≥77%), with much lower intra-subject and technical replicate variability. The relatively large degree of inter-subject variation is apparent from mean and standard deviation values for MN-RET (0.15 ± 0.10%) and MUT RET (4.7 ± 5.0 per million, after omission of two extreme outliers). The influences of age and sex on inter-subject variation were investigated, and neither factor affected MN-RET whereas both influenced MUT RET frequency. The lowest MUT RET values were observed for subjects <11 years old, and males had moderately higher frequencies than females. These results indicate that MN-RET and MUT RET are automation-compatible biomarkers of genotoxicity that bridge species of toxicological interest to include human populations. These data will be useful for appropriately designing future human studies that include these biomarkers of genotoxicity, and highlight the need for additional work aimed at identifying the sources of inter-individual variability reported herein.
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影响因子：
3.8
作者：
Guerard, Melanie;Koenig, Julie;Zeller, Andreas
通讯作者：
Zeller, Andreas
影响因子：
4.8
作者：
Offer, T;Ho, E;Ames, BN
通讯作者：
Ames, BN
PIG-A 基因突变作为人群研究中的基因毒性生物标志物：对铅暴露工人的调查。
影响因子：
2.8
作者：
Cao, Yiyi;Wang, Tuanwei;Luan, Yang
通讯作者：
Luan, Yang
DOI：
10.2307/2987937
发表时间：
1983-01-01
期刊：
JOURNAL OF THE ROYAL STATISTICAL SOCIETY SERIES D-THE STATISTICIAN
影响因子：
--
作者：
ALTMAN, DG;BLAND, JM
通讯作者：
BLAND, JM
影响因子：
4.6
作者：
Haboubi, Hasan N.;Lawrence, Rachel L.;Jenkins, Gareth J.
通讯作者：
Jenkins, Gareth J.