Campana D, Janossy G
Blood. 1986 Dec;68(6):1264-71.
In this paper a microplate method is described for diagnosing acute leukemia and for investigating the reactivity of monoclonal antibodies (MoAbs) against membrane antigens in combination with rabbit or murine antibodies to nuclear terminal transferase (TdT). The speed of this method facilitates the investigation of fresh leukemic cells from individual patients and assesses the cytolytic efficacy of the relevant MoAbs in the presence of complement (C'). Lymphoblasts (TdT+) are mixed in equal proportions with known numbers of "inert" cells, eg, RBC or nonleukemic bone marrow (BM). Following incubation with MoAbs and C' the ratio of residual TdT+ cells and inert cells is determined on cytospin preparations. Initially, percentages of TdT+ cells are counted in a unit volume of 5,000 inert cells, followed by the scanning of greater than 2 X 10(4) inert cells on entire slides. With this method more than 4 log cytoreduction of TdT + cells is detected. The method is also applicable for studying the cytolysis of malignant B cells by using mostly monoclonal lg expression rather than TdT for the identification of residual B cells. Ten representative patients selected from a group of greater than 100 are reported. In some cases cytoreduction of greater than 4 log with no identifiable residual TdT + cells is achieved by a single C'-fixing MoAb: anti-CD10 (RFAL3) in common acute lymphoid leukemia (ALL) and anti-CD7 (RFT2) in T cell ALL (T-ALL). Other cases require cocktails of anti-CD10, anti-CD19, and anti-CD24 in common ALL or anti-CD7 and anti-CD8 in T-ALL. In T-ALL a few TdT + cells remain that exhibit the features of normal TdT + BM cells (CD7-, HLA-DR+). This is particularly noticeable when patients are studied in partial remission or if nonleukemic BM is used as a source of inert cells. The methods described here contribute to establishing a range of MoAbs (ie, of IgM class) and techniques for efficient purging and to comparing the efficacy of "clean-up," in remission, of common ALL, T-ALL, and B cell malignancies.
本文描述了一种微孔板方法，用于诊断急性白血病，并研究单克隆抗体（MoAb）与兔或鼠抗核末端转移酶（TdT）抗体联合作用于膜抗原的反应性。该方法的速度便于对个体患者的新鲜白血病细胞进行研究，并在补体（C'）存在的情况下评估相关MoAb的细胞溶解效力。将淋巴母细胞（TdT+）与已知数量的“惰性”细胞（如红细胞或非白血病骨髓（BM））按等比例混合。在用MoAb和C'孵育后，在细胞涂片制备物上测定残留的TdT+细胞与惰性细胞的比例。最初，在5000个惰性细胞的单位体积中计数TdT+细胞的百分比，随后在整个载玻片上扫描超过2×10⁴个惰性细胞。用这种方法可检测到TdT+细胞超过4个对数级的细胞减少。该方法也适用于通过主要利用单克隆Ig表达而非TdT来鉴定残留B细胞，从而研究恶性B细胞的细胞溶解。报告了从100多名患者中选出的10名代表性患者。在某些情况下，单一的补体固定MoAb：普通急性淋巴细胞白血病（ALL）中的抗CD10（RFAL3）和T细胞ALL（T-ALL）中的抗CD7（RFT2）可实现超过4个对数级的细胞减少且无可识别的残留TdT+细胞。其他情况则需要普通ALL中的抗CD10、抗CD19和抗CD24鸡尾酒或T-ALL中的抗CD7和抗CD8鸡尾酒。在T-ALL中，仍有一些TdT+细胞保留正常TdT+ BM细胞的特征（CD7-，HLA-DR+）。当在部分缓解期研究患者或使用非白血病BM作为惰性细胞来源时，这种情况尤为明显。本文所述方法有助于建立一系列MoAb（即IgM类）和有效清除技术，并比较普通ALL、T-ALL和B细胞恶性肿瘤在缓解期“清除”的疗效。
