van Kooten C, Rensink I, Aarden L, van Oers R
Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
Leukemia. 1993 Oct;7(10):1576-84.
We studied the in vitro differentiation (immunoglobulin production) of purified malignant B cells of 21 patients with different B-cell malignancies, including chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), hairy cell leukemia (HCL) and non-Hodgkin lymphoma (NHL). Direct activation of purified malignant B cells with phorbol myristate acetate (PMA) resulted in the differentiation of most CLL cells, but not of the other types of B-cell malignancies. This differentiation required the presence of interleukin 4 (IL-4). In contrast, with the use of anti-CD2-stimulated normal T cells and IL-2, immunoglobulin M (IgM) could be detected in the supernatant of all but one of the purified malignant B-cell populations. However, by analysis of the light chains of the IgM produced, monoclonality could be demonstrated in only 13/21 cases: 8/11 CLL, 3/3 PLL, 0/3 HCL, and 2/4 NHL. In two patients additional proof that the malignant B cells were the source of the IgM production could be obtained in an idiotype-specific ELISA. Apart from IgM, also the production of IgG antibodies could be detected. However, only for 2/3 HCL patients, we could confirm a monoclonal IgG production. Since HCL is a malignancy of mature B cells, already carrying IgG on the membrane, this IgG production is not the result of a switch process. In all other cases where IgG production was polyclonal, we have no indications for the induction of Ig switch. The fact that the more mature B-cell malignancies were T-cell-dependent for their differentiation might be a reflection of the in-vivo situation. The efficient induction of malignant B-cell differentiation described in this paper allows investigation of the antigen specificity of these antibodies.
我们研究了21例不同B细胞恶性肿瘤患者纯化的恶性B细胞的体外分化（免疫球蛋白产生）情况，这些患者包括慢性淋巴细胞白血病（CLL）、原淋巴细胞白血病（PLL）、毛细胞白血病（HCL）和非霍奇金淋巴瘤（NHL）。用佛波酯（PMA）直接激活纯化的恶性B细胞可导致大多数CLL细胞分化，但其他类型的B细胞恶性肿瘤细胞则无此现象。这种分化需要白细胞介素4（IL-4）的存在。相比之下，使用抗CD2刺激的正常T细胞和IL-2时，除一个纯化的恶性B细胞群体外，在所有其他群体的上清液中均可检测到免疫球蛋白M（IgM）。然而，通过分析所产生IgM的轻链，仅在13/21例中证实存在单克隆性：11例CLL中有8例、3例PLL均为3例、3例HCL均为0例、4例NHL中有2例。在两名患者中，通过独特型特异性酶联免疫吸附测定（ELISA）可进一步证明恶性B细胞是IgM产生的来源。除IgM外，还可检测到IgG抗体的产生。然而，仅在2/3的HCL患者中，我们能够证实存在单克隆IgG产生。由于HCL是成熟B细胞的恶性肿瘤，其细胞膜上已携带IgG，因此这种IgG产生并非转换过程的结果。在所有其他IgG产生为多克隆的情况下，我们没有发现Ig转换诱导的迹象。更成熟的B细胞恶性肿瘤在分化上依赖T细胞这一事实可能反映了体内情况。本文所述的恶性B细胞分化的有效诱导使得能够研究这些抗体的抗原特异性。
