In vitro immunogenic profile of recombinant SARS-CoV2 S1-RBD peptide in murine macrophage and microglial cells.

IF 2.5 4区 医学 Q2 PARASITOLOGY Memorias do Instituto Oswaldo Cruz Pub Date : 2023-01-01 DOI:10.1590/0074-02760220144
Adriano José Maia Chaves Filho, Paloma Marinho Jucá, Michelle Verde Ramo Soares, Caio Andrade de Oliveira, Raul Cavalcante de Sousa, Deniele Bezerra Lós, Remo Castro Russo, Juliana Navarro Ueda Yaochite, Danielle S Macedo
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Abstract

Background: The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants can infect common mice inducing significant pathological lung lesions and inflammatory responses. This substantially mimics coronavirus disease 19 (COVID-19) infection and pathogenesis in humans.

Objectives: To characterise the effects of recombinant SARS-CoV-2 S1 receptor-binding domain (RBD) peptide in murine macrophage and microglial cells' immune activation compared with classical PAMPs in vitro.

Methods: Murine RAW 264.7 macrophages and BV2 microglial cells were exposed to increasing concentrations of the RBD peptide (0.01, 0.05, and 0.1 µg/mL), Lipopolysaccharide (LPS) and Poly(I:C) and evaluated after two and 24 h for significant markers of macrophage activation. We determined the effects of RBD peptide on cell viability, cleaved caspase 3 expressions, and nuclear morphometry analysis.

Findings: In RAW cells, RBD peptide was cytotoxic, but not for BV2 cells. RAW cells presented increased arginase activity and IL-10 production; however, BV2 cells expressed iNOS and IL-6 after RBD peptide exposure. In addition, RAW cells increased cleaved-caspase-3, apoptosis, and mitotic catastrophe after RBD peptide stimulation but not BV2 cells.

Conclusion: RBD peptide exposure has different effects depending on the cell line, exposure time, and concentration. This study brings new evidence about the immunogenic profile of RBD in macrophage and microglial cells, advancing the understanding of SARS-Cov2 immuno- and neuropathology.

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重组SARS-CoV2 S1-RBD肽在小鼠巨噬细胞和小胶质细胞中的体外免疫原性分析。
背景:新型严重急性呼吸综合征冠状病毒2 (SARS-CoV-2)可感染普通小鼠,引起明显的病理肺病变和炎症反应。这基本上模拟了冠状病毒病19 (COVID-19)在人类中的感染和发病机制。目的:研究重组SARS-CoV-2 S1受体结合域(RBD)肽对小鼠巨噬细胞和小胶质细胞免疫激活的影响,并与经典PAMPs进行比较。方法:将小鼠RAW 264.7巨噬细胞和BV2小胶质细胞分别暴露于浓度增加的RBD肽(0.01、0.05和0.1µg/mL)、脂多糖(LPS)和Poly(I:C)中,分别在2 h和24 h后评估巨噬细胞活化的显著标志。我们测定了RBD肽对细胞活力、裂解caspase 3表达和核形态分析的影响。结果:在RAW细胞中,RBD肽具有细胞毒性,但对BV2细胞无细胞毒性。RAW细胞精氨酸酶活性和IL-10产量增加;然而,RBD肽暴露后,BV2细胞表达了iNOS和IL-6。此外,RBD肽刺激后,RAW细胞增加了裂解caspase-3、凋亡和有丝分裂突变,而BV2细胞则没有。结论:RBD肽暴露对不同细胞系、暴露时间和暴露浓度有不同的影响。本研究为巨噬细胞和小胶质细胞中RBD的免疫原性谱提供了新的证据,促进了对SARS-Cov2免疫和神经病理学的认识。
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来源期刊
CiteScore
5.00
自引率
3.60%
发文量
91
审稿时长
3-8 weeks
期刊介绍: Memórias do Instituto Oswaldo Cruz is a journal specialized in microbes & their vectors causing human infections. This means that we accept manuscripts covering multidisciplinary approaches and findings in the basic aspects of infectious diseases, e.g. basic in research in prokariotes, eukaryotes, and/or virus. Articles must clearly show what is the main question to be answered, the hypothesis raised, and the contribution given by the study. Priority is given to manuscripts reporting novel mechanisms and general findings concerning the biology of human infectious prokariotes, eukariotes or virus. Papers reporting innovative methods for diagnostics or that advance the basic research with these infectious agents are also welcome. It is important to mention what we do not publish: veterinary infectious agents research, taxonomic analysis and re-description of species, epidemiological studies or surveys or case reports and data re-analysis. Manuscripts that fall in these cases or that are considered of low priority by the journal editorial board, will be returned to the author(s) for submission to another journal.
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