Modak M J, Mertelsmann R, Koziner B, Pahwa R, Moore M A, Clarkson B D, Good R A
J Cancer Res Clin Oncol. 1980;98(1):91-104. doi: 10.1007/BF00413181.
A micromethod for the determination of TdT in peripheral leukocytes and bone marrow cells has been developed that allows unequivocal identification and quantitation of TdT in less than 1 X 10(6) leukocytes from ALL patients, i.e., in 1 ml of peripheral blood and/or 0.5 ml of bone marrow obtained during routine clinical sampling. The method involves disruption of cell pellet with high salt and detergent followed by centrifugation of extracts at 12,000 X g and partial purification on phosphocellulose matrix by a batch elution technique using a standard laboratory microcentrifuge. Using this microassay, TdT activities have been determined in 500 samples of peripheral blood and bone marrow of 240 adult patients with acute leukemias (86 ALL, 108 ANLL, 44 blastic CML, two acute leukemias following P. vera). From an analysis of our data based on TdT activity, cell surface markers and growth patterns in soft agar and observations published in the literature, it can be concluded that the frequencies of TdT + phenotypes in the various clinical-morphological diagnostic groups are approximately 95% in ALL, 10% in ANLL, 50% in AUL, and 35% in blastic CML. Since the presence of high TdT activity is clearly associated with clinical response to specific forms of chemotherapy in blastic CML and most probably, also in ANLL, the determination of TdT should be considered in all cases of acute leukemias to objectively define prognostically important subgroups which can not be diagnosed by conventional means.
已开发出一种用于测定外周血白细胞和骨髓细胞中末端脱氧核苷酸转移酶（TdT）的微量方法，该方法能够在来自急性淋巴细胞白血病（ALL）患者的少于1×10⁶个白细胞中，即在1毫升外周血和/或常规临床采样时获取的0.5毫升骨髓中，明确鉴定和定量TdT。该方法包括用高盐和去污剂破坏细胞沉淀，然后以12,000×g离心提取物，并使用标准实验室微量离心机通过批量洗脱技术在磷酸纤维素基质上进行部分纯化。使用这种微量测定法，已对240例成年急性白血病患者（86例ALL、108例急性非淋巴细胞白血病（ANLL）、44例急变期慢性粒细胞白血病（CML）、2例真性红细胞增多症后急性白血病）的500份外周血和骨髓样本测定了TdT活性。根据我们基于TdT活性、细胞表面标志物以及软琼脂中的生长模式的数据分析以及文献中发表的观察结果，可以得出结论，在各种临床 - 形态学诊断组中，TdT⁺表型的频率在ALL中约为95%，在ANLL中为10%，在急性未分化白血病（AUL）中为50%，在急变期CML中为35%。由于高TdT活性的存在显然与急变期CML以及很可能在ANLL中对特定形式化疗的临床反应相关，因此在所有急性白血病病例中都应考虑测定TdT，以客观地定义通过传统方法无法诊断的具有重要预后意义的亚组。
