狂犬病是由狂犬病病毒（RABV）引起的一种可预防的人畜共患疾病，死亡率很高。信使RNA（mRNA）疫苗为疫苗开发和大流行防范开辟了新途径，具有强大的可扩展性，可能克服唯一获得许可的狂犬病灭活疫苗浪费时间和成本的缺点。在这里，我们设计了一种表达 RABV G 蛋白并用脂质纳米颗粒 (LNP) 和不同的核酸免疫刺激剂 (CPG 1018、CPG 2395 和 Poly I:C) 封装的 RABV mRNA 疫苗，然后评估其在小鼠中的免疫原性和保护能力。虽然用 LNP 和 CPG 1018 封装的 RABV mRNA 可以诱导更有效的体液反应，具有最高和持久的 RABV-G 特异性 IgG 滴度和病毒中和滴度，但也诱导更强的 RABV G 特异性细胞介导的免疫 (CMI) 反应，包括最高的根据小鼠流式细胞术测定，产生干扰素 γ (IFN-γ) 和肿瘤坏死因子 α (TNFα) 的 CD4 /CD8 T 细胞的比例。此外，在暴露前和暴露后攻击试验中，LNP CPG 1018 封装的 RABV G mRNA 可诱导 100% 针对 25 LD50 的 RABV 感染，通过 qRT-PCR 检测到病毒基因组减少，病毒复制抑制效果最高。这些结果表明，用 LNP 免疫刺激核酸 CPG 1018 封装的 RABV G mRNA 显示出作为安全且经济的狂犬病候选疫苗的前景。版权所有 © 2023 作者。由爱思唯尔有限公司出版。保留所有权利。

Rabies is a preventable zoonotic disease caused by rabies virus (RABV) with high mortality. Messenger RNA (mRNA) vaccines have opened up new avenues for vaccine development and pandemic preparedness with potent scalability, which may overcome the only licensed rabies inactived vaccine' shortcoming of time and cost wasting. Here, we designed an RABV mRNA vaccines expressed RABV G protein and capsulated with lipid nanoparticle (LNP) and different nucleic acid immunostimulator (CPG 1018, CPG 2395 and Poly I:C) and then assessed the immunogenicity and protective capacity in mice. While RABV mRNA capsulated with LNP and CPG 1018 could induce more potent humoral response with highest and durable RABV-G specific IgG titers and virus neutralizing titers, but also induced stronger RABV G-specific cell-mediated immunity (CMI) responses, including the highest proportions of interferon-γ (IFN-γ) and tumor necrosis factor alpha (TNFα)- producing CD4+/CD8 + T cells according to a flow cytometry assay in mice. In addition, in the pre- and post-exposure challenge assays, LNP + CPG 1018 capsulated RABV G mRNA induced 100 % protection against 25 LD50 of RABV infection with highest inhibition efficacy of viral replication with the decreased virus genome detected by qRT-PCR. These results showed that RABV G mRNA capsulated with LNP immune-stimulating nucleic acids CPG 1018 showed promise as a safe and economical rabies vaccine candidate.Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.