Generation of cytotoxic T lymphocytes by MHC class I ligands fused to heat shock cognate protein 70
Generation of cytotoxic T lymphocytes by MHC class I ligands fused to heat shock cognate protein 70
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DOI：
10.1093/intimm/13.10.1233
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发表时间：
2001-10-01
影响因子：
4.4
通讯作者：
Yui, K
中科院分区：
文献类型：
--
作者：
Udono, H;Yamano, T;Yui, K
用gp96和热休克同源蛋白70(Hsc70)免疫小鼠，可以诱导肽特异性细胞毒性T淋巴细胞(CTL)，其中hsc70由体内天然多肽结合或体外重组合成的多肽结合而成。此外，分枝杆菌热休克蛋白70与卵清蛋白(OVA)衍生片段的共价融合已被证明能产生MHC I类限制性CTL反应。在这里，我们将约氏疟原虫环子孢子蛋白、肿瘤抗原、HY抗原和OVA五种不同的CTL表位基因融合到小鼠hsc70的N-端或C-端，并在大肠杆菌中表达。所有5种融合蛋白免疫后均可诱导出多肽特异性CTL，表明CTL表位的同源侧翼区域不是免疫应答所必需的。注射部位是诱导CTL的关键。CD8(+)T细胞的启动不需要CD4(+)T细胞，用hsc70融合蛋白冲击的骨髓来源的树突状细胞免疫也能诱导CTL反应。此外，通过hsc70的缺失突变体，我们确定hsc70的280-385氨基酸残基是诱导CTL反应最关键的区域。
Immunization with gp96 and heat shock cognate protein 70 (hsc70) purified with in vivo bound naturally occurring peptides or bound to synthetic peptides by in vitro reconstitution has been shown to induce peptide-specific cytotoxic T lymphocytes (CTL). In addition, mycobacterial heat shock protein 70 covalently fused to ovalbumin (OVA)-derived fragments has been shown to generate MHC class I-restricted CTL responses. Here, we genetically fused five different CTL epitopes, including peptides derived from Plasmodium yoelli circumsporozoite protein, tumor antigens, HY antigen and OVA, to either the N- or C-terminus of murine hsc70 and expressed the resulting proteins in Escherichia coli. Vaccination with all five fusion proteins induced peptide-specific CTL, indicating that no cognate flanking regions of CTL epitopes are necessary for the immune response. The point of injection was crucial for CTL induction. CD4(+) T cells were not required for the priming of CD8(+) T cells and vaccination with bone marrow-derived dendritic cells pulsed with hsc70 fusion proteins also elicited CTL responses. Furthermore, by using deletion mutants of hsc70, we identified amino acid residues 280-385 of hsc70 as the region most critical for inducing the CTL response.