A Single-Chain H-2Db Molecule Presenting an Influenza Virus Nucleoprotein Epitope Shows Enhanced Ability at Stimulating CD8+ T Cell Responses In Vivo
A Single-Chain H-2Db Molecule Presenting an Influenza Virus Nucleoprotein Epitope Shows Enhanced Ability at Stimulating CD8+ T Cell Responses In Vivo
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DOI：
10.4049/jimmunol.0803893
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发表时间：
2009-04-15
影响因子：
4.4
通讯作者：
Gould, Keith G.
中科院分区：
文献类型：
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作者：
Palmowski, Michael J.;Parker, Mathew;Gould, Keith G.
我们已经生成了编码单链H-2D（B）小鼠MHC I类分子的构建体，其中流感病毒核蛋白（NP）表位（氨基酸序列ASNENMDAM）通过柔性接头序列与小鼠β（2）-微球蛋白和D-b融合。B H链。这种单链三聚体（SCT）在细胞表面有效表达，不依赖于TAP和内源性β（2）-微球蛋白，并在体外被特异性T细胞直接有效识别。编码D-b NP SCT的重组牛痘病毒在C57 BL/6小鼠中引发的CD 8（+）T细胞应答比表达NP表位作为小基因的等效病毒高4倍，如四聚体染色所示，无论小基因是否通过信号序列定向到内质网中。这种反应是功能性的，如肽脉冲靶细胞的体内裂解试验所示，并且在用流感病毒体内二次攻击后大大扩增。在使用F5 TCR转基因脾细胞的过继转移实验中，SCT对CD 8（+）细胞的免疫刺激性也显著高于NP小基因，其中T细胞应答的幅度要大得多。我们的结果扩展了以前使用SCT的DNA疫苗接种研究，该研究表明这些分子能够产生功能性CD 8（+）T细胞应答。我们已经证明，I类SCT的免疫原性甚至比表位小基因形式的预处理抗原更强，因此，当需要产生最佳CD 8（+）T细胞应答时，应考虑使用I类SCT。免疫学杂志，2009，182：4565-4571.
We have generated a construct encoding a single-chain H-2D(b) mouse MHC class I molecule in which an influenza virus nucleoprotein (NP) epitope, amino acid sequence ASNENMDAM, is fused to mouse beta(2)-microglobulin and the D-b H chain via flexible linker sequences. This single-chain trimer (SCT) was efficiently expressed at the cell surface independently of TAP and endogenous beta(2)-microglobulin, and it was recognized directly and efficiently by specific T cells in vitro. A recombinant vaccinia virus encoding the D-b NP SCT primed a CD8(+) T cell response in C57BL/6 mice 4-fold greater than an equivalent virus expressing the NP epitope as a minigene, as shown by tetramer staining, whether or not the minigene was directed into the endoplasmic reticulum by a signal sequence. This response was functional as shown by in vivo lysis assays with peptide-pulsed target cells, and it was greatly expanded following secondary challenge in vivo with influenza virus. The SCT was also significantly more immunostimulatory for CD8(+) cells than the NP minigene in adoptive transfer experiments using F5 TCR transgenic spleen cells, in which the magnitude of the T cell response was much greater. Our results extend previous DNA vaccination studies using SCTs, which demonstrated that such molecules are capable of generating functional CD8(+) T cell responses. We have shown that class I SCTs are more immunogenic than even preprocessed Ag in the form of an epitope minigene, and they therefore should be considered for use when the generation of optimal CD8(+) T cell responses is required. The Journal of Immunology, 2009, 182: 4565-4571.