Moritz D, Groner B
Institute for Experimental Cancer Research, Tumor Biology Center, Freiburg, Germany.
Gene Ther. 1995 Oct;2(8):539-46.
The elimination of tumor cells by cytotoxic T lymphocytes (CTLs) could become the basis of a biological cancer therapy. The recognition specificity of cytotoxic T lymphocytes (CTLs) can be genetically modified by stable introduction of chimeric T cell receptor genes and thus be directed towards tumor cells. We designed a recombinant T cell receptor (TCR) component consisting of a single chain Fv derivative of a monoclonal antibody (scFv) serving as the extracellular antigen-binding domain and the zeta-chain of the TCR/CD3 complex serving as a signal transducing domain. Three chimeric receptor constructs differing in their molecular structure were derived and their functions in transduced T cells compared. A construct in which the scFv domain, specific for the ErbB-2 receptor, was fused directly to the zeta-chain, and two constructs containing different hinge regions between the functional domains, were made. The hinge regions serve as spacers which increase the distance of the scFv moiety from the plasma membrane. Only the two scFv-zeta chimeras containing a hinge region showed ErbB-2 binding activity, when expressed in T cells. The scFv-zeta construct which lacks a spacer segment did not. Consistently, only the spacer-containing chimeras transduced T cell receptor signals following ErbB-2 mediated crosslinking. An increase in intracellular Ca2+ concentrations and cytokine secretion was observed. ErbB-2 expressing tumor cells were efficiently lysed by CTLs which expressed the spacer-containing scFv-zeta chimeras. Our results will help to optimize the design of recombinant T cell receptor components useful in the grafting of a specificity of recognition on to cytotoxic T cells and possibly the gene therapy of cancer.
细胞毒性T淋巴细胞（CTL）对肿瘤细胞的清除作用可能成为生物癌症治疗的基础。通过稳定导入嵌合T细胞受体基因，可对细胞毒性T淋巴细胞（CTL）的识别特异性进行基因改造，从而使其靶向肿瘤细胞。我们设计了一种重组T细胞受体（TCR）组件，其由作为细胞外抗原结合域的单克隆抗体（scFv）的单链Fv衍生物和作为信号转导域的TCR/CD3复合物的ζ链组成。我们得到了三种分子结构不同的嵌合受体构建体，并比较了它们在转导T细胞中的功能。构建了一种将对ErbB-2受体具有特异性的scFv结构域直接与ζ链融合的构建体，以及两种在功能域之间包含不同铰链区的构建体。铰链区起到间隔物的作用，可增加scFv部分与质膜之间的距离。当在T细胞中表达时，只有两种包含铰链区的scFv-ζ嵌合体表现出ErbB-2结合活性。缺少间隔片段的scFv-ζ构建体则没有。一致地，只有包含间隔区的嵌合体在ErbB-2介导的交联后转导T细胞受体信号。观察到细胞内Ca2+浓度增加和细胞因子分泌。表达ErbB-2的肿瘤细胞被表达含间隔区scFv-ζ嵌合体的CTL有效裂解。我们的结果将有助于优化重组T细胞受体组件的设计，这些组件可用于将识别特异性移植到细胞毒性T细胞上，并可能用于癌症的基因治疗。
