Schindler D, Kubbies M, Priest R E, Hoehn H, Rabinovitch P S
Department of Human Genetics, University of Würzburg, F.R.G.
Mech Ageing Dev. 1988 Sep;44(3):253-63. doi: 10.1016/0047-6374(88)90026-7.
Mononuclear cells were isolated from peripheral blood by a standard Ficoll-Hypaque technique from 127 healthy donors, ranging in age from newborns to 86 years of age. As a measure of their in vitro growth response, the fraction of non-cycling cells was determined at 48 and 72 h after phytohemagglutinin (PHA) exposure by means of BrdU-Hoechst flow cytometry. This technique provides an optimal assay system for the non-cycling cell fraction, since all cycling cells will have incorporated BrdU thereby quenching the fluorescence of the Hoechst 33258 fluorochrome. Lymphocytes from prepubertal donors showed significantly decreased non-cycling cell fractions, as did lymphocytes from an additional group of 14 adults with hypogonadism due to the 45, XO condition (Turner-Syndrome). Much to our surprise, we found no definitive correlation between donor age and the non-cycling fraction of cells from the adult lymphocyte donors. Nor did we find any age-related increase in the variance of the non-cycling cell fraction. These observations suggest that the previously reported age-related decline in the PHA response of human PBL may reflect an increasing delay, rather than an overall diminution, of the PHA response as a function of donor age.
通过标准的Ficoll-泛影葡胺技术从127名年龄从新生儿到86岁的健康供体的外周血中分离出单核细胞。作为其体外生长反应的一种衡量指标，在暴露于植物血凝素（PHA）后48小时和72小时，通过BrdU-赫斯特流式细胞术测定非循环细胞的比例。由于所有循环细胞都会掺入BrdU，从而淬灭赫斯特33258荧光染料的荧光，因此该技术为非循环细胞比例提供了一个最佳的检测系统。青春期前供体的淋巴细胞显示非循环细胞比例显著降低，因45,XO状况（特纳综合征）导致性腺功能减退的另外一组14名成年人的淋巴细胞也是如此。令我们惊讶的是，我们发现供体年龄与成年淋巴细胞供体细胞的非循环比例之间没有明确的相关性。我们也没有发现非循环细胞比例的方差随年龄有任何增加。这些观察结果表明，先前报道的人类外周血淋巴细胞（PBL）PHA反应随年龄下降可能反映了PHA反应作为供体年龄的函数，延迟增加，而不是整体减弱。
