Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan,
Cytotechnology. 2006 Nov;52(3):171-9. doi: 10.1007/s10616-006-9035-2. Epub 2006 Dec 5.
IL-6 has been known to modulate the growth of many hybridoma cells and also promote resultant antibody productivity. However, IL-6 is so expensive that the use of IL-6-dependent hybridomas for industrial antibody production is not practical. In this study, we aimed at designing antibody/gp130 and antibody/EpoR chimeras which could tightly control cell growth in response to more affordable cognate antigen. Retroviral vectors encoding V(H) or V(L) region of anti-hen egg lysozyme (HEL) antibody HyHEL-10 tethered to a pair of extracellular D2/transmembrane domains of erythropoietin receptor (EpoR) and cytoplasmic domains of either EpoR or gp130, were constructed, and a homodimeric or a heterodimeric pair of chimeric receptor combinations (V(H)-gp130 and V(L)-gp130 or V(H)-gp130 and V(L)-EpoR) were expressed in an IL-6-dependent hybridoma 7TD1. The chimeric receptor-derived growth signal was observed in both combinations, while some residual growth signal was observed in the absence of HEL. To reduce interchain interaction between the two receptor chains, we introduced mutations to the transmembrane domain of both chimera combinations. Consequently, the heterodimeric combination of V(H)-gp130 and V(L)-EpoR showed clear HEL-dependent cell growth, while the homodimeric combination of V(H)-gp130 and V(L)-gp130 showed reduced cell growth in the absence of HEL. This is the first report that an EpoR-gp130 cytoplasmic domain heterodimer could transduce a growth signal in hybridoma cells, indicating tight and economical growth control of hybridoma cells via our chimeric receptors.
白细胞介素 6（IL-6）已被证实能够调节许多杂交瘤细胞的生长，并促进相应抗体的产生。然而，IL-6 非常昂贵，因此使用依赖于 IL-6 的杂交瘤进行工业抗体生产是不切实际的。在本研究中，我们旨在设计抗体/gp130 和抗体/EpoR 嵌合体，使其能够根据更经济实惠的同源抗原严格控制细胞生长。构建了编码抗鸡卵溶菌酶（HEL）抗体 HyHEL-10 的 V(H)或 V(L)区的逆转录病毒载体，该抗体通过一对红细胞生成素受体（EpoR）的胞外 D2/跨膜结构域和 EpoR 或 gp130 的胞质结构域连接，表达在依赖于白细胞介素 6 的杂交瘤 7TD1 中。在这两种嵌合受体组合（V(H)-gp130 和 V(L)-gp130 或 V(H)-gp130 和 V(L)-EpoR）中均观察到了嵌合受体衍生的生长信号，而在没有 HEL 的情况下也观察到了一些残留的生长信号。为了减少两条受体链之间的链间相互作用，我们对两种嵌合体组合的跨膜结构域都进行了突变。结果，V(H)-gp130 和 V(L)-EpoR 的异源二聚体组合表现出明显依赖于 HEL 的细胞生长，而 V(H)-gp130 和 V(L)-gp130 的同源二聚体组合在没有 HEL 的情况下细胞生长减少。这是第一个报道 EpoR-gp130 胞质结构域异源二聚体能够在杂交瘤细胞中传递生长信号的报告，表明通过我们的嵌合受体可以对杂交瘤细胞进行严格和经济的生长控制。
