Subtle affinity-enhancing mutations in a myelin oligodendrocyte glycoprotein-specific TCR alter specificity and generate new self-reactivity.
Subtle affinity-enhancing mutations in a myelin oligodendrocyte glycoprotein-specific TCR alter specificity and generate new self-reactivity.
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DOI：
10.4049/jimmunol.0804377
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发表时间：
2009-04-01
期刊：
影响因子：
--
通讯作者：
Geiger TL
中科院分区：
文献类型：
--
作者：
Udyavar A;Alli R;Nguyen P;Baker L;Geiger TL
我们描述了一个简单的迭代方法来增加TCR的亲和力，我们研究使用髓鞘少突胶质细胞糖蛋白（MOG）特异性TCR。我们假设T细胞受体（TCR）CDR 3中的单个氨基酸修饰可以通过与抗原肽的局部相互作用增强TCR敏感性，同时最大限度地降低先前在使用体外进化技术更广泛诱变的TCR中观察到的交叉反应性风险。我们表明，这种迭代方法确实可以产生抗原敏感性比亲本受体高100倍的TCR，并且可以赋予TCR与共受体的独立性。然而，我们还发现，CDR 3中的单个氨基酸突变可以改变TCR精细特异性，从而影响在MHC结合沟的大部分长度上对抗原残基的识别要求。此外，表面暴露的CDR 3氨基酸的最小变化，甚至单个羟基的添加或甲基或巯基部分转化为羟基，可以赋予修饰的抗原特异性TCR新的自身反应性。通过将逆转录病毒TCR基因转移到Rag−/−小鼠中，对经修饰的TCR进行体内建模，证实了这些改变的反应性的生物学意义，尽管也证明了产生抗原特异性、正选择性、共受体非依赖性受体的可行性，其抗原敏感性显著增加。这些结果证实了容易产生用于治疗目的的亲和力增强的TCR的可能性，但证明了TCR CDR 3结构的最小变化可以促进自身反应性，从而强调了在临床应用之前验证具有甚至细微改变的受体时谨慎的重要性。
We describe a simple iterative approach to augment TCR affinity, which we studied using a myelin oligodendroglial glycoprotein (MOG)-specific TCR. We hypothesized that single amino acid modifications in T-cell receptor (TCR) CDR3 could enhance TCR sensitivity through focal interactions with antigenic peptide while minimizing the risk of cross-reactivity observed previously in TCRs more broadly mutagenized using in vitro evolution techniques. We show that this iterative method can indeed generate TCR with antigen sensitivity 100-fold greater than the parental receptor, and can endow TCR with co-receptor independence. However, we also find that single amino acid mutations in the CDR3 can alter TCR fine specificity, affecting recognition requirements for antigen residues over most of the length of the MHC binding groove. Further, minimal changes in surface-exposed CDR3 amino acids, even the addition of a single hydroxyl group or conversion of a methyl or sulfhydryl moiety to a hydroxyl, can confer modified antigen-specific TCR with new self reactivity. In vivo modeling of modified TCR through retroviral TCR gene transfer into Rag−/− mice confirmed the biological significance of these altered reactivities, though also demonstrated the feasibility of producing antigen-specific, positively selecting, co-receptor independent receptors with markedly increased antigen sensitivity. These results affirm the possibility of readily generating affinity-enhanced TCR for therapeutic purposes, but demonstrate that minimal changes in TCR CDR3 structure can promote self reactivity and thereby emphasize the importance of caution in validating receptors with even subtle alterations prior to clinical application.
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