Srivastava B I, Minowada J
Leuk Res. 1983;7(3):331-8. doi: 10.1016/0145-2126(83)90097-8.
We have examined alterations in terminal deoxynucleotidyl transferase (TdT) immunofluorescence (IF) in MOLT-4 cells during changes in growth conditions. Subsequently, we used cells from 48 human hematopoietic cell lines of different cell lineages and maturation stages to compare the IF and biochemical assays for expression of TdT. In addition, we have attempted to correlate the expression of TdT, adenosine deaminase (ADA), thymidine phosphorylase (TP) and immunological markers with maturation stages in these different cell lines. The results indicate that TdT positive cells remain TdT positive when assayed by either the biochemical or IF tests during growth or early plateau phase, but that cells under poor growth conditions, such as in old cultures, may give a negative TdT IF reaction. Otherwise, biochemical and IF assays for TdT gave comparable results in the 48 cell lines tested, testifying to the reliability of the IF test. Based on the comparisons of the various cell lines studied, it appears that both ADA and TdT decrease progressively as maturation of T cells from Blast I to Blast IV to mature T cells increases. TP was deficient in all T-cell lines compared to normal peripheral blood T cells, which in turn had lower activity compared to normal peripheral blood B cells. Pre-B cells, although indistinguishable from each other by immunological markers and all having low TP and ADA activity, showed heterogeneity, with TdT activity high in some and low in others. All non-T, non-B lines had high TdT activity, but low ADA and TP activity. B- and myelocytic cell lines had low ADA and TdT activity, and showed an increase in TP activity as the maturation of cells increased. These results indicate that the TdT IF test is a reliable procedure for detecting TdT positive cells, and that TdT, ADA and TP could be useful markers for studying the differentiation of human hematopoietic cells.
我们研究了在生长条件改变期间MOLT-4细胞中末端脱氧核苷酸转移酶(TdT)免疫荧光(IF)的变化。随后,我们使用来自48种不同细胞谱系和成熟阶段的人类造血细胞系的细胞,比较TdT表达的IF检测和生化检测。此外,我们试图将TdT、腺苷脱氨酶(ADA)、胸苷磷酸化酶(TP)的表达以及免疫标志物与这些不同细胞系的成熟阶段相关联。结果表明,在生长或早期平台期通过生化或IF检测时,TdT阳性细胞仍为TdT阳性,但处于生长条件较差的细胞,如传代较久的培养物中,可能会给出TdT IF阴性反应。否则,在测试的48种细胞系中,TdT的生化检测和IF检测结果相当,证明了IF检测的可靠性。基于对所研究的各种细胞系的比较,似乎随着T细胞从原始I期到原始IV期再到成熟T细胞的成熟度增加,ADA和TdT均逐渐降低。与正常外周血T细胞相比,所有T细胞系中的TP均缺乏,而正常外周血T细胞与正常外周血B细胞相比活性又较低。前B细胞虽然通过免疫标志物彼此无法区分,且均具有低TP和ADA活性,但表现出异质性,一些细胞的TdT活性高,而另一些细胞的TdT活性低。所有非T、非B细胞系均具有高TdT活性,但ADA和TP活性低。B细胞系和髓细胞系具有低ADA和TdT活性,并且随着细胞成熟度增加,TP活性升高。这些结果表明,TdT IF检测是检测TdT阳性细胞的可靠方法,并且TdT、ADA和TP可能是研究人类造血细胞分化的有用标志物。