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含有铁蛋白笼状纳米结构和变异链球菌葡糖基转移酶葡聚糖结合区的自组装抗龋粘膜疫苗可有效预防啮齿动物龋齿形成。

Self-assembling anticaries mucosal vaccine containing ferritin cage nanostructure and glucan-binding region of S. mutans glucosyltransferase effectively prevents caries formation in rodents.

机构信息

a The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine of Ministry of Education (KLOBM) , School & Hospital of Stomatology, Wuhan University , Wuhan , China.

b Department of Endodontics , School and Hospital of Stomatology, Wuhan University , Wuhan , China.

出版信息

Hum Vaccin Immunother. 2017 Oct 3;13(10):2332-2340. doi: 10.1080/21645515.2017.1349046.

Abstract

Anticaries protein vaccines that induce a mucosal immune response are not effective. Therefore, development of effective and convenient anticaries vaccines is a priority of dental research. Here we generated self-assembling nanoparticles by linking the glucan-binding region of Streptococcus mutans glucosyltransferase (GLU) to the N-terminal domain of ferritin to determine whether these novel nanoparticles enhanced the immunogenicity of an anticaries protein vaccine against GLU in rodents. We constructed the expression plasmid pET28a-GLU-FTH and purified the proteins from bacteria using size-exclusion chromatography. BALB/c mice were used to evaluate the ability of GLU-ferritin (GLU-FTH) nanoparticles to induce GLU-specific mucosal and systemic responses. The protective efficiency of GLU-FTH nanoparticles was compared with that of GLU alone or a mixture of GLU and poly(I:C) after administering an intranasal infusion to Wistar rats. The phagocytosis and maturation of dendritic cells (DCs) exposed in vitro to the nanoparticles were assessed using flow cytometry. The GLU-FTH nanoparticle vaccine elicited significantly higher levels of GLU-specific antibodies compared with GLU or a mixture of GLU and poly(I:C). Immunization with GLU-FTH achieved lower caries scores compared with those of the other vaccines. Administration of GLU-FTH nanoparticles enhanced phagocytosis by DCs and their maturation. Thus, self-assembling GLU-FTH is a highly effective anticaries mucosal vaccine that enhanced antibody production and inhibited S. mutans infection in rodents.

摘要

抗龋蛋白疫苗诱导黏膜免疫反应的效果并不理想。因此,开发有效且方便的抗龋疫苗是口腔医学研究的重点。在这里,我们通过将变异链球菌葡聚糖转移酶 (GLU) 的葡聚糖结合区与铁蛋白的 N 端结构域相连,生成了自组装纳米颗粒,以确定这些新型纳米颗粒是否能增强 GLU 抗龋蛋白疫苗在啮齿动物中的免疫原性。我们构建了表达质粒 pET28a-GLU-FTH,并使用排阻层析法从细菌中纯化了蛋白质。使用 BALB/c 小鼠评估 GLU-铁蛋白 (GLU-FTH) 纳米颗粒诱导 GLU 特异性黏膜和全身反应的能力。将 GLU-FTH 纳米颗粒的保护效率与 GLU 单独或 GLU 与聚肌苷酸 (poly(I:C)) 混合物经鼻内滴注给药后在 Wistar 大鼠中的效果进行了比较。使用流式细胞术评估了体外暴露于纳米颗粒的树突状细胞 (DC) 的吞噬作用和成熟情况。与 GLU 或 GLU 与 poly(I:C) 混合物相比,GLU-FTH 纳米疫苗诱导的 GLU 特异性抗体水平显著升高。与其他疫苗相比,GLU-FTH 免疫接种的龋齿评分较低。GLU-FTH 纳米颗粒的给药增强了 DC 的吞噬作用及其成熟。因此,自组装的 GLU-FTH 是一种高效的抗龋黏膜疫苗,可增强抗体产生并抑制变形链球菌在啮齿动物中的感染。

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