Vaccine T-cell epitope selection by a peptide competition assay.
Vaccine T-cell epitope selection by a peptide competition assay.
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通过肽竞争测定进行疫苗 T 细胞表位选择。
DOI：
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发表时间：
1989
影响因子：
11.1
通讯作者：
F. Sinigaglia
中科院分区：
文献类型：
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作者：
J. Kilgus;P. Romagnoli;M. Guttinger;D. Stuber;L. Adorini;F. Sinigaglia
在竞争测定中检查了源自恶性疟原虫环子孢子蛋白（CS 蛋白）的几种肽与人类主要组织相容性复合物 II 类蛋白 HLA-DR5 和 -DRw6 的结合。将固定的抗原呈递细胞 (APC) 与不同浓度的每种肽和次优浓度的刺激肽一起孵育。然后使用两个已建立的 DR5 或 DRw6 限制性 T 细胞克隆（对刺激肽具有特异性）作为应答细胞，在增殖测定中确定 CS 肽与 DR5 或 DRw6 蛋白的结合。发现五种 CS 肽之一（总共包含约 50% 的 CS 蛋白序列）与刺激肽与 DR5 和 DRw6 的结合竞争。然后，与 DR5 和 DRw6 结合的 CS 肽 CS-(378-398) 被证明能够诱导来自具有 DR5 和 DRw6 单倍型的供体的 T 细胞的初次体外反应。在适当的 APC 存在下，CS-(378-398) 诱导的 T 细胞克隆不仅对同源肽有反应，而且对天然 CS 蛋白也有反应。我们所采用的策略对于针对任何病原体的疫苗工程具有相当大的普遍兴趣，因为它极大地促进了选择合适的 T 细胞表位以纳入疫苗中。
The binding of several peptides derived from the Plasmodium falciparum circumsporozoite protein (CS protein) to the human major histocompatibility complex class II proteins HLA-DR5 and -DRw6 was examined in a competition assay. Fixed antigen-presenting cells (APCs) were incubated with various concentrations of each peptide and suboptimal concentrations of stimulator peptides. The binding of the CS peptides to DR5 or DRw6 proteins was then determined in a proliferation assay using two established DR5 or DRw6-restricted T-cell clones with specificity for the stimulator peptides as responder cells. One of five CS peptides, comprising together about 50% of the CS protein sequence, was found to compete with the binding of the stimulator peptides to DR5 and DRw6. The CS peptide CS-(378-398), binding to DR5 and DRw6, was then shown to be able to induce primary in vitro responses of T cells from donors with DR5 and DRw6 haplotypes. CS-(378-398)-induced T-cell clones responded not only to the homologous peptide but also to the native CS protein in the presence of appropriate APCs. The strategy we have applied is of considerable general interest for the engineering of vaccines against any pathogen, since it greatly facilitates the selection of appropriate T-cell epitopes to be incorporated in the vaccine.