中文摘要：本研究组成员在先前的研究中构建了一个免疫耐受弹性蛋白样多肽(iTEPs)载体用于传递CTL疫苗，且增强了疫苗的作用。为进一步优化疫苗载体的作用，本研究构建了一个能够传递佐剂和疫苗表位的免疫耐受弹性蛋白样多肽载体。结果表明，iTEPs与H-100融合能够在体外诱导树突状细胞（DCs）的成熟，这可以通过CD80和CD86表达量的增加得到验证。iTEPs与H-100融合同时能够通过增加促炎性细胞因子IL-6的分泌来促进DCs的激活。无论是H-100的C末端还是T末端与iTEP融合，都能表现出相同的作用，融合后通过减少丝氨酸的结构来激活DC。iTEP融合物能够在体内外增强抗原特异性CTL反应，从而可以作为CTL的佐剂进行应用。当H-100-iTEP与CTL表位结合后能够形成一个单分子疫苗，这种自带佐剂作用的疫苗与携带不完整的弗氏佐剂的疫苗相比，能够诱导产生更强烈的抗原特异性CTL反应。
外文摘要：Cytotoxic T lymphocyte (CTL) epitope peptide-based vaccines are widely used in cancer and infectious disease therapy. We previously generated an immune-tolerant elastin-like polypeptides (iTEPs)-based carrier to deliver a peptide CTL vaccine and enhance the efficiency of the vaccine. To further optimize the vaccine carrier, we intended to potentiate its function by designing an iTEP-based carrier that was able to deliver adjuvant and a vaccine epitope as one molecule. Thus, we fused a 9-mer H-100, a peptide derived from the high-mobility group box 1 protein (HMGB1) that could induce activation of dendritic cells (DCs), with an iTEP polymer to generate a new iTEP polymer named H-100-iTEP. The H-100-iTEP still kept the feature of reversible phase transition of iTEPs and should be able to be used as a polymer carrier to deliver peptide vaccines. The expression levels of CD80/CD86 on DCs were assessed using flow cytometry. The iTEP fusion-stimulated IL-6 secretion by DCs was measured with ELISA. Activation of antigen-specific CD8+ T cells induced by iTEP fusions was examined through a B3Z hybridoma cell activation assay. In vivo CTL activation promoted by iTEP fusions was detected by an IFN-gamma-based ELISPOT assay. The iTEP fused with H-100 could induce maturation of DCs in vitro as evidenced by increased CD80 and CD86 expression. The iTEP fusion also promoted activation of DCs by increasing secretion of a proinflammatory cytokine IL-6. The N-terminus or C-terminus fusion of H-100 to iTEP had a similar effect and a reduced form of cysteine in iTEP fusions was required for DC stimulation. iTEP fusions potentiated a co-administrated CTL vaccine by increasing an antigen-specific CTL response in vitro and in vivo. When the H-100-iTEP was fused to a CTL epitope to generate a one-molecule vaccine, this self-adjuvanted vaccine elicited a stronger antigen-specific CTL response than a vaccine adjuvanted by Incomplete Freund's Adjuvant. Thus, we have successfully generated a functional, one-molecule iTEP-based self-adjuvanted vaccine.
外文关键词：iTEP； CTL vaccine； peptide adjuvant； dendritic cell activation/maturation； self-adjuvanted vaccine； CTL response
作者： Dong, SY; Xu, TF; Wang, P; Zhao, P; Chen, MN
作者单位：美国犹他大学
期刊名称：ACTA PHARMACOLOGICA SINICA
期刊影响因子：3.223
出版年份：2017
出版刊次：6
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