Numasaki M, Nakamura K, Fukuoka Y, Saeki H, Hanai N, Kudo T
Cancer Cell Repository, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Miyagi, Japan.
Immunol Lett. 2001 Jan 15;75(3):167-77. doi: 10.1016/s0165-2478(00)00308-4.
A human monoclonal antibody (HuMab) TONO-1 (IgM, lambda) recognizes cell surface antigens associated primarily with human T-leukemia/lymphoma cells. In this study, we investigated the reactivity against T-leukemia/lymphoma cells in detail, cytotoxic potential and primary nucleotide and deduced amino acid sequences of the rearranged heavy and light chains of the HuMab TONO-1. Expression of the molecules (TONO-1 Ags) detected by a HuMab TONO-1 was significantly heterogeneous even in the same T-leukemia/lymphoma cell lines HPB-MLT and MOLT-4F. The flow cytometric curves showed an unusual broad-based spread of fluorescence intensity. HuMab TONO-1 was shown to have the ability to kill the T-leukernia/lymphoma cells efficiently in the presence of rabbit complements. However, HuMab TONO-1 did not demonstrate significant antibody-dependent cellular cytotoxic activity. Furthermore, HuMab TONO-1 heavy and light chain variable regions were cloned, sequenced and analyzed. HuMab TONO-1 uses a V(H) gene member of the V(H)IV gene family V(H)71-4, and is productively rearranged with the germ line D(H) gene D(XP')1, and the germ line J(H)5 gene with multiple somatic mutations. HuMab TONO-1 Vlambda belongs to the lambda light chain variable subgroup I family and is derived from the Vlambdalc germ line gene Humlv1042, and germ line gene Jlambda1 without somatic mutations. The results reveal that the production of HuMab TONO-1, with cytotoxic potential for human T-leukemia/lymphoma cells, is achieved by rearrangement of the V(H)71-4/Humlv1042 germ line variable region gene combination, that is associated with the autoimmune repertoire.
一种人源单克隆抗体（HuMab）TONO-1（IgM，λ）可识别主要与人T淋巴细胞白血病/淋巴瘤细胞相关的细胞表面抗原。在本研究中，我们详细研究了其对T淋巴细胞白血病/淋巴瘤细胞的反应性、细胞毒性潜力以及HuMab TONO-1重链和轻链重排后的初级核苷酸序列和推导的氨基酸序列。即使在同一T淋巴细胞白血病/淋巴瘤细胞系HPB-MLT和MOLT-4F中，由HuMab TONO-1检测到的分子（TONO-1抗原）的表达也存在显著异质性。流式细胞术曲线显示荧光强度呈现异常的宽基分布。结果表明，在兔补体存在的情况下，HuMab TONO-1能够有效杀伤T淋巴细胞白血病/淋巴瘤细胞。然而，HuMab TONO-1并未表现出显著的抗体依赖性细胞毒性活性。此外，对HuMab TONO-1重链和轻链可变区进行了克隆、测序和分析。HuMab TONO-1使用V(H)IV基因家族V(H)71-4的一个V(H)基因成员，与种系D(H)基因D(XP')1以及具有多个体细胞突变的种系J(H)5基因进行了有效重排。HuMab TONO-1 Vλ属于λ轻链可变亚组I家族，源自种系基因Vlambdalc Humlv1042，且种系基因Jλ1未发生体细胞突变。结果表明，具有杀伤人类T淋巴细胞白血病/淋巴瘤细胞细胞毒性潜力的HuMab TONO-1是通过V(H)71-4/Humlv1042种系可变区基因组合的重排产生的，这与自身免疫库相关。
