Enhanced Humoral and Cellular Immune Responses Elicited by Adenoviral Delivery of SARS-CoV-2 Receptor-Binding Motif Fused to Human Fc.

IF 5.2 3区 医学 Q1 IMMUNOLOGY Vaccines Pub Date : 2024-11-01 DOI:10.3390/vaccines12111247
Yea-Jin Lee, Maheswaran Easwaran, Yong-Sam Jung, Yingjuan Qian, Hyun-Jin Shin
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Abstract

Background/Objectives: The receptor binding motif (RBM) of the SARS-CoV-2 spike protein is critical for viral entry into host cells. Development of a vaccine targeting this region is a promising strategy for COVID-19 prevention. To enhance the immunogenicity of SARS-CoV-2 vaccines, we developed an adenoviral vector expressing the RBM from the SARS-CoV-2 spike protein that fused to the human Fc (hFc) domain. Methods: The recombinant RBM_hFc fusion protein was successfully cloned into the pacAd5CMV-N-pA (pAd5) vector and expressed in HEK293 cells as a ~40 kDa protein. A recombinant adenovirus encoding RBM_hFc was subsequently generated and confirmed by cytopathic effect assay. Results: Western blot analysis verified the expression of RBM_hFc in the adenovirus (AdV). ELISA assays, validated for IgG detection, demonstrated a twofold increase in IgG antibody levels (M-1.090 at 450 nm; SD-±0.326; and 95% CI-0.250 [0.839 to 1.340]) in sera from BALB/c mice immunized with Ad/RBM_hFc, compared to the negative control group. Result suggests a robust humoral immune response induced by the Ad/RBM_hFc vaccine. Moreover, ELISpot assays demonstrated a tenfold increase in IFN-γ -producing cells (M-440 spot-forming cells; SD-±124.976; and 95% CI-75.522 [364.478 to 515.522]) in mice immunized with AdV/RBM_hFc compared to the negative control group. Result proved that AdV/RBM_hFc-stimulated a robust cellular immune response in animal model. Conclusions: Our findings indicate that the RBM_hFc fusion protein enhances both humoral and cellular immune responses. These results suggest the potential of adenoviral vectors carrying RBM_hFc as vaccine candidates. However, comprehensive evaluation of the protective efficacy of these adenoviral vectors will necessitate rigorous experimental studies.

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腺病毒递送与人 Fc 融合的 SARS-CoV-2 受体结合基团可增强体液和细胞免疫反应
背景/目的:SARS-CoV-2 穗状病毒蛋白的受体结合基团 (RBM) 是病毒进入宿主细胞的关键。开发针对该区域的疫苗是预防 COVID-19 的一种有前途的策略。为了提高 SARS-CoV-2 疫苗的免疫原性,我们开发了一种表达 SARS-CoV-2 尖峰蛋白 RBM 的腺病毒载体,该载体与人 Fc(hFc)结构域融合。方法:重组 RBM_hFc 融合蛋白成功克隆到 pacAd5CMV-N-pA (pAd5)载体中,并在 HEK293 细胞中表达为 ~40 kDa 蛋白。随后生成了编码 RBM_hFc 的重组腺病毒,并通过细胞病理效应实验进行了确认。结果Western 印迹分析验证了 RBM_hFc 在腺病毒 (AdV) 中的表达。经IgG检测验证的ELISA检测表明,与阴性对照组相比,用Ad/RBM_hFc免疫的BALB/c小鼠血清中的IgG抗体水平增加了两倍(450 nm波长下M-1.090;SD-±0.326;95% CI-0.250 [0.839至1.340])。结果表明,Ad/RBM_hFc 疫苗诱导了强大的体液免疫反应。此外,ELISpot 试验表明,与阴性对照组相比,免疫 AdV/RBM_hFc 的小鼠产生 IFN-γ 的细胞(M-440 形成斑点的细胞;SD-±124.976;和 95% CI-75.522 [364.478 至 515.522])增加了十倍。结果证明,在动物模型中,AdV/RBM_hFc能激发强有力的细胞免疫反应。结论:我们的研究结果表明,RBM_hFc 融合蛋白能增强体液免疫和细胞免疫反应。这些结果表明了携带 RBM_hFc 的腺病毒载体作为候选疫苗的潜力。不过,要全面评估这些腺病毒载体的保护效力,还需要进行严格的实验研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Vaccines
Vaccines Pharmacology, Toxicology and Pharmaceutics-Pharmacology
CiteScore
8.90
自引率
16.70%
发文量
1853
审稿时长
18.06 days
期刊介绍: Vaccines (ISSN 2076-393X) is an international, peer-reviewed open access journal focused on laboratory and clinical vaccine research, utilization and immunization. Vaccines publishes high quality reviews, regular research papers, communications and case reports.
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