TCR-engineered T cells: a model of inducible TCR expression to dissect the interrelationship between two TCRs.
TCR-engineered T cells: a model of inducible TCR expression to dissect the interrelationship between two TCRs.
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DOI：
10.1002/eji.201343591
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发表时间：
2014-01
影响因子：
5.4
通讯作者：
Uckert, Wolfgang
中科院分区：
文献类型：
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作者：
Reuss, Simone;Sebestyen, Zsolt;Heinz, Niels;Loew, Rainer;Baum, Christopher;Debets, Reno;Uckert, Wolfgang
TCR基因修饰的过继治疗T细胞同时表达TGTCR和内源性TCR，可能导致TCR错配，导致TCR-TGT细胞功能下降。我们在两种情况下产生了双TCR T细胞，其中一种TCR由逆转录病毒启动子组成性表达，而第二种TCR表达由Tet-On系统调节。组成性表达的TCR分子在细胞表面的表达依赖于诱导的TCR表达而减少，导致功能严重受阻。此外，利用荧光共振能量转移技术，我们检测到错配的TCR二聚体和不同的TCR链的配对行为，并相互影响TCR链的表达。改变TCR的表达水平或引入额外的半胱氨酸桥并不能避免功能丧失和错配。然而，在多克隆T细胞中，优化的TCR形式(半胱氨酸化、密码子优化)增强了正确的配对和功能。我们从我们的数据中得出结论：(I)错配水平取决于单个TCR，并且不会随着一个TCR水平的增加而降低，(Ii)修饰(半胱氨化、密码子优化)可以改善正确的配对，但不能完全排除错配(半胱氨酸化)。
TCR gene modified T cells for adoptive therapy simultaneously express the Tg TCR and the endogenous TCR, which might lead to mispaired TCRs with harmful unknown specificity and to a reduced function of TCR-Tg T cells. We generated dual TCR T cells in two settings in which either TCR was constitutively expressed by a retroviral promoter while the second TCR expression was regulable by a Tet-on system. Constitutively expressed TCR molecules were reduced on the cell surface depending on the induced TCR expression leading to strongly hampered function. Besides that, using fluorescence resonance energy transfer we detected mispaired TCR dimers and different pairing behaviors of individual TCR chains with a mutual influence on TCR chain expression. The loss of function and mispairing could not be avoided by changing the TCR expression level or by introduction of an additional cysteine bridge. However, in polyclonal T cells, optimized TCR formats (cysteineization, codon optimization) enhanced correct pairing and function. We conclude from our data that (i) the level of mispairing depends on the individual TCRs and is not reduced by increasing the level of one TCR, and (ii) modifications (cysteineization, codon optimization) improve correct pairing but do not completely exclude mispairing (cysteineization).
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影响因子：
56.9
作者：
Morgan, Richard A.;Dudley, Mark E.;Rosenberg, Steven A.
通讯作者：
Rosenberg, Steven A.
影响因子：
8.6
作者：
Scholten, KBJ;Kramer, D;Hooijberg, E
通讯作者：
Hooijberg, E
影响因子：
4.4
作者：
Schüler, T;Blankenstein, T
通讯作者：
Blankenstein, T
影响因子：
45.3
作者：
Robbins, Paul F.;Morgan, Richard A.;Rosenberg, Steven A.
通讯作者：
Rosenberg, Steven A.
影响因子：
5.1
作者：
通讯作者：
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