Minimal Amino Acid Exchange in Human TCR Constant Regions Fosters Improved Function of TCR Gene-Modified T Cells
Minimal Amino Acid Exchange in Human TCR Constant Regions Fosters Improved Function of TCR Gene-Modified T Cells
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DOI：
10.4049/jimmunol.0902055
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发表时间：
2010-06-01
影响因子：
4.4
通讯作者：
Uckert, Wolfgang
中科院分区：
文献类型：
--
作者：
Sommermeyer, Daniel;Uckert, Wolfgang
过继转移TCR基因修饰的T细胞进行TCR基因治疗是治疗肿瘤的一种新策略。TCR基因治疗的一个关键前提是充分表达所转移的TCR。人们开发了几种策略来实现最佳表达，包括用小鼠对应物取代人类TCRα和TCRβ恒定区的“小鼠化”。使用一系列人-鼠杂交构建物，我们已经确定了九种氨基酸，这些氨基酸与鼠化TCR表达的改善有关。在TCRβC区确定了5种必需氨基酸交换，其中最重要的是C区第18位的谷氨酸(人)与碱性赖氨酸(鼠)的交换。对于TCRαC区，四个氨基酸的面积足以提高表达。最低限度的鼠化TCR变异体(仅含有9个小鼠序列残基)通过支持转移的TCR链的优先配对和与CD3蛋白更稳定的结合而增强了人TCR的表达。最重要的是，与野生型TCR转导细胞相比，使用最低限度的鼠化TCR链改善了转导的原代人类T细胞的功能。对于TCR基因治疗，使用最少的恒定区而不是完全杀灭的恒定区大大减少了外源残基的数量，从而降低了治疗性TCR的免疫原性风险。免疫学杂志，2010,184：6223-6231。
TCR gene therapy using adoptive transfer of TCR gene-modified T cells is a new strategy for treatment of cancer. One critical prerequisite for TCR gene therapy is sufficient expression of transferred TCRs. Several strategies to achieve optimal expression were developed, including "murinization," which replaces the human TCR alpha and TCR beta constant regions by their murine counterparts. Using a series of mouse-human hybrid constructs, we have identified nine amino acids responsible for the improved expression of murinized TCRs. Five essential amino acid exchanges were identified in the TCR beta C region, with exchange of a glutamic acid (human) for a basic lysine (mouse) at position 18 of the C region, being most important. For the TCR alpha C region, an area of four amino acids was sufficient for improved expression. The minimally murinized TCR variants (harboring only nine residues of the mouse sequence) enhanced expression of human TCRs by supporting preferential pairing of transferred TCR chains and a more stable association with the CD3 proteins. Most important, usage of minimally murinized TCR chains improved the function of transduced primary human T cells in comparison with cells transduced with wild-type TCRs. For TCR gene therapy, the utilization of minimally instead of completely murinized constant regions dramatically reduces the number of foreign residues and thereby the risk for immunogenicity of therapeutic TCRs. The Journal of Immunology, 2010, 184: 6223-6231.