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基于 DNA 适体的滚环扩增产物作为一种新型免疫佐剂。

DNA aptamer-based rolling circle amplification product as a novel immunological adjuvant.

机构信息

Department of Cell and Molecular Biology, University of Arkansas, Fayetteville, AR, 72701, USA.

Department of Medical Laboratory Techniques, Kut Tech Institute, Middle Technical University, Baghdad, Iraq.

出版信息

Sci Rep. 2020 Dec 17;10(1):22282. doi: 10.1038/s41598-020-79420-w.

DOI:10.1038/s41598-020-79420-w
PMID:33335251
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7747709/
Abstract

Several agonists to CD40 have shown to induce acquired immune responses. Here, we developed and evaluated the rolling circle amplification (RCA) products that are based on anti-CD40 DNA aptamers as a novel vaccine adjuvant. First, we developed DNA aptamers with specific binding affinity to chicken CD40 extra domain (chCD40ED). Next, we prepared the RCA products that consist of these aptamers to increase the spanning space and overall binding affinity to chCD40ED. Using 8 DNA aptamer candidates, 4 aptamer-based RCA products (aptamer RCAs) were generated, each consisting of two distinct aptamers. We demonstrated that all 4 aptamer RCAs significantly induced the signal transduction in chicken HD11 macrophage cell line (p < 0.05). Finally, we conjugated one of the aptamer RCAs (Aptamer RCA II) to M2e epitope peptide of influenza virus as a model hapten, and the immune complex was injected to chickens. Aptamer RCA II stimulated anti-M2e IgG antibody production to the level significantly higher as compared to the control (M2e epitope alone; p < 0.05). The results of our work suggest that aptamer RCA is a novel platform to boost the efficacy of vaccines, which might find broad applications to other antigens beyond M2e epitope evaluated in this study using chicken infection model.

摘要

几种 CD40 激动剂已被证明可诱导获得性免疫反应。在这里，我们开发并评估了基于抗 CD40 DNA 适体的滚环扩增 (RCA) 产物，作为一种新型疫苗佐剂。首先，我们开发了与鸡 CD40 额外结构域 (chCD40ED) 具有特异性结合亲和力的 DNA 适体。接下来，我们制备了由这些适体组成的 RCA 产物，以增加跨度空间和对 chCD40ED 的整体结合亲和力。使用 8 个 DNA 适体候选物，生成了 4 种基于适体的 RCA 产物（适体 RCA），每种产物都由两个不同的适体组成。我们证明，所有 4 种适体 RCA 均显著诱导鸡 HD11 巨噬细胞系的信号转导（p<0.05）。最后，我们将一种适体 RCA（Aptamer RCA II）与流感病毒的 M2e 表位肽缀合作为模型半抗原，并将免疫复合物注射到鸡中。与对照（单独的 M2e 表位；p<0.05）相比，Aptamer RCA II 刺激抗 M2e IgG 抗体产生的水平显著更高。我们的工作结果表明，适体 RCA 是一种增强疫苗效力的新型平台，该平台可能会在使用鸡感染模型评估的本研究之外的其他抗原中找到广泛的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/8d50936e87f1/41598_2020_79420_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/42b4bac5d423/41598_2020_79420_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/b12d28613934/41598_2020_79420_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/44331ce37f6c/41598_2020_79420_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/0f8fa7a4f81c/41598_2020_79420_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/8d50936e87f1/41598_2020_79420_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/42b4bac5d423/41598_2020_79420_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/b12d28613934/41598_2020_79420_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/44331ce37f6c/41598_2020_79420_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/0f8fa7a4f81c/41598_2020_79420_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375e/7747709/8d50936e87f1/41598_2020_79420_Fig5_HTML.jpg

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