Voss Ralf-Holger, Willemsen Ralph A, Kuball Jürgen, Grabowski Margarete, Engel Renate, Intan Ratna S, Guillaume Philippe, Romero Pedro, Huber Christoph, Theobald Matthias
Medical Clinic (III) and Polyclinic, Department of Hematology and Oncology, Johannes Gutenberg-University, Mainz, Germany.
J Immunol. 2008 Jan 1;180(1):391-401. doi: 10.4049/jimmunol.180.1.391.
A promising approach to adoptive transfer therapy of tumors is to reprogram autologous T lymphocytes by TCR gene transfer of defined Ag specificity. An obstacle, however, is the undesired pairing of introduced TCRalpha- and TCRbeta-chains with the endogenous TCR chains. These events vary depending on the individual endogenous TCR and they not only may reduce the levels of cell surface-introduced TCR but also may generate hybrid TCR with unknown Ag specificities. We show that such hybrid heterodimers can be generated even by the pairing of human and mouse TCRalpha- and TCRbeta-chains. To overcome this hurdle, we have identified a pair of amino acid residues in the crystal structure of a TCR that lie at the interface of associated TCR Calpha and Cbeta domains and are related to each other by both a complementary steric interaction analogous to a "knob-into-hole" configuration and the electrostatic environment. We mutated the two residues so as to invert the sense of this interaction analogous to a charged "hole-into-knob" configuration. We show that this inversion in the CalphaCbeta interface promotes selective assembly of the introduced TCR while preserving its specificity and avidity for Ag ligand. Noteworthily, this TCR modification was equally efficient on both a Mu and a Hu TCR. Our data suggest that this approach is generally applicable to TCR independently of their Ag specificity and affinity, subset distribution, and species of origin. Thus, this strategy may optimize TCR gene transfer to efficiently and safely reprogram random T cells into tumor-reactive T cells.
肿瘤过继性转移治疗的一种有前景的方法是通过转导具有特定抗原特异性的TCR基因来重编程自体T淋巴细胞。然而，一个障碍是引入的TCRα链和TCRβ链与内源性TCR链发生不期望的配对。这些事件因个体内源性TCR的不同而有所差异，它们不仅可能降低细胞表面引入的TCR水平，还可能产生具有未知抗原特异性的杂交TCR。我们发现，即使是人源和鼠源的TCRα链和TCRβ链配对也能产生这种杂交异二聚体。为了克服这一障碍，我们在TCR的晶体结构中确定了一对氨基酸残基，它们位于相关TCR的Cα和Cβ结构域的界面处，通过类似于“旋钮插入孔洞”构型的互补空间相互作用以及静电环境相互关联。我们对这两个残基进行了突变，以反转这种相互作用的方向，类似于带电的“孔洞插入旋钮”构型。我们发现，CαCβ界面的这种反转促进了引入的TCR的选择性组装，同时保留了其对抗原配体的特异性和亲和力。值得注意的是，这种TCR修饰对鼠源和人源TCR同样有效。我们的数据表明，这种方法普遍适用于TCR，而与其抗原特异性、亲和力、亚群分布及来源物种无关。因此，这种策略可能会优化TCR基因转导，从而高效且安全地将随机T细胞重编程为肿瘤反应性T细胞。
