Single and dual amino acid substitutions in TCR CDRs can enhance antigen-specific T cell functions
Single and dual amino acid substitutions in TCR CDRs can enhance antigen-specific T cell functions
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DOI：
10.4049/jimmunol.180.9.6116
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发表时间：
2008-05-01
影响因子：
4.4
通讯作者：
Rosenberg, Steven A.
中科院分区：
文献类型：
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作者：
Robbins, Paul F.;Li, Yong F.;Rosenberg, Steven A.
在识别肿瘤相关Ags的三种TCR的TCR CDRs中产生了单氨基酸和双氨基酸替代变体。利用基于rna的快速转染系统鉴定了增强TCR基因修饰T细胞对同源Ag复合物反应性的替代。在HLA-A*02 I类等位基因背景下，1G4 TCR的一组变体识别与人癌睾丸Ag NY-ESO-1（SLLMWITQC）氨基酸残基157-165相对应的肽，结果鉴定出单和双CDR3 α和CDR2 β氨基酸替代，显著增强了TCR基因修饰的CD4(+) T细胞对NY-ESO-1(+)/HLA-A*02（+）肿瘤细胞系的特异性识别。在这组改进的TCR中，在1G4 TCR CDR3a链上发现了双重替代，增强了基因修饰的CD4（+）和CD8(+) T细胞中的ag特异性反应性。在两种不同的tcr上进行的单独实验表明，识别mar -1 27-35 （AAGIGILTV）肽/HLA-A*02 Ag复合物的tcr中，单氨基酸取代可以增强CD4(+) T细胞的Ag特异性反应性。这些结果表明，可以通过快速转染和分析技术鉴定增强T细胞功能的简单TCR替代变体，为生成有效的Ag复合物特异性TCR基因提供了手段，用于研究T细胞相互作用和T细胞过用性免疫治疗。
Single and dual amino acid substitution variants were generated in the TCR CDRs of three TCRs that recognize tumor-associated Ags. Substitutions that enhance the reactivity of TCR gene-modified T cells to the cognate Ag complex were identified using a rapid RNA-based transfection system. The screening of a panel of variants of the 1G4 TCR, that recognizes a peptide corresponding to amino acid residues 157-165 of the human cancer testis Ag NY-ESO-1 (SLLMWITQC) in the context of the HLA-A*02 class I allele, resulted in the identification of single and dual CDR3 alpha and CDR2 beta amino acid substitutions that dramatically enhanced the specific recognition of NY-ESO-1(+)/HLA-A*02(+) tumor cell lines by TCR gene-modified CD4(+) T cells. Within this group of improved TCRs, a dual substitution in the 1G4 TCR CDR3a chain was identified that enhanced Ag-specific reactivity in gene-modified CD4(+) and CD8(+) T cells. Separate experiments on two distinct TCRs that recognize the MART-1 27-35 (AAGIGILTV) peptide/HLA-A*02 Ag complex characterized single amino acid substitutions in both TCRs that enhanced CD4(+) T cell Ag-specific reactivity. These results indicate that simple TCR substitution variants that enhance T cell function can be identified by rapid transfection and assay techniques, providing the means for generating potent Ag complex-specific TCR genes for use in the study of T cell interactions and in T cell adoptive immunotherapy.