Collste L G, Darzynkiewicz Z, Traganos F, Sharpless T K, Whitmore W F, Melamed M R
Cancer Res. 1979 Jun;39(6 Pt 1):2120-4.
The reactivity of lymphocytes in lymph nodes draining the site of a transplantable experimental bladder tumor (MBT2 in C3H/HeJ mice) has been measured in a multiparameter flow cytometry system. Acridine orange was used as a nucleic acid probe. This dye intercalates in helical DNA, emitting green (530 nm) fluorescence upon exposure to blue light; it stacks to single-stranded RNA, emitting red (640 nm) fluorescence. The relative magnitude of the increase of lymphocyte DNA and RNA has been evaluated simultaneously in tumor-draining nodes, in nondraining nodes of the same animal, and in untreated control animals. Stimulation of the regional node lymphocytes could be observed after 20 days but not after 10 days. It was uniformly high at 35 days. The transcriptive response (increased proportion of lymphocytes with high RNA) was more pronounced than the proliferative (increased proportion of lymphocytes with more than diploid DNA). The histological changes in the stimulated nodes resembled closely those described by others in human tumor-draining nodes. The described method has the advantage of being simple, rapid, and able to measure a representative part of the whole-cell population.
在多参数流式细胞术系统中，对可移植性实验性膀胱肿瘤（C3H/HeJ小鼠中的MBT2）部位引流淋巴结中的淋巴细胞反应性进行了测量。吖啶橙用作核酸探针。这种染料嵌入螺旋DNA中，在蓝光照射下发出绿色（530nm）荧光；它堆积在单链RNA上，发出红色（640nm）荧光。同时评估了肿瘤引流淋巴结、同一动物的非引流淋巴结和未处理对照动物中淋巴细胞DNA和RNA增加的相对幅度。20天后可观察到区域淋巴结淋巴细胞受到刺激，但10天后未观察到。在35天时一直很高。转录反应（高RNA淋巴细胞比例增加）比增殖反应（二倍体以上DNA淋巴细胞比例增加）更明显。受刺激淋巴结的组织学变化与其他人描述的人类肿瘤引流淋巴结的变化非常相似。所描述的方法具有简单、快速且能够测量全细胞群体代表性部分的优点。
