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Designing the epitope flanking regions for optimal generation of CTL epitopes

机译：设计表位侧翼区域以最佳生成CTL表位

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摘要

The flanking amino acids that surround epitopes are critical for effective antigen processing and maintenance of epitope integrity. In the present study, the frequency and characteristics of each amino acid that flanked the peptides generated from the proteasomal degradation of three different subtypes of HIV-1 Gag-p24 were determined. Synthetic flanking regions were designed based on the highest and the lowest frequencies of amino acid with the ideal characteristics at positions upstream and downstream of the proteasomal cleavage site. Peptides were synthesized that contained known CD8+ CTL-epitopes from HIV-1 Gag, CMV pp65, and vaccinia proteins HRP-2, and C16, flanked by amino acid sequences specifically designed to either generate or inhibit the known CD8+ CTL-epitopes. As predicted, the known CD8+ CTL-epitopes were effectively generated from the peptides with synthetic flanking regions specifically designed to promote epitope generation in a proteasome-dependent manner. The majority of the proteasome-generated epitopes were cleaved immediately after the C-terminal amino acid of the specific CTL-epitope. The synthetic peptide sequences containing known CD8+ CTL-epitopes with the flanking regions that promote epitope generation were effectively processed and presented to epitope specific CD8+ T-cells resulting in the production of IFN-gamma. These results highlight the importance of flanking regions in promoting efficient antigen processing and presentation. This concept can have important implications in vaccine design and development strategies
机译：围绕表位的侧翼氨基酸对于有效的抗原加工和表位完整性的维持至关重要。在本研究中，确定了从三种不同亚型的HIV-1 Gag-p24的蛋白酶体降解产生的肽两侧的每个氨基酸的频率和特征。基于在蛋白酶体切割位点上游和下游位置具有理想特性的氨基酸的最高和最低频率设计合成侧翼区。合成的肽包含来自HIV-1 Gag，CMV pp65和牛痘蛋白HRP-2和C16的已知CD8 + CTL表位，其侧翼是专门设计用于生成或抑制已知CD8 + CTL表位的氨基酸序列。如所预测的，已知的CD8 + CTL-表位是从具有合成侧翼区的肽有效产生的，所述侧翼区经特别设计以以蛋白酶体依赖性方式促进表位的产生。蛋白酶体生成的大多数表位在特定CTL表位的C端氨基酸后立即被切割。含有已知的具有促进表位生成的侧翼区的CD8 + CTL-表位的合成肽序列被有效地加工并呈递给表位的特异性CD8 + T细胞，从而产生IFN-γ。这些结果突出了侧翼区在促进有效抗原加工和呈递中的重要性。该概念可能对疫苗设计和开发策略产生重要影响

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