Laboratory of Atherothrombosis, Cardiology Department, Federal State Budgetary Educational Institution of Higher Education (FSBEI HE) "Russian University of Medicine" of the Ministry of Health of the Russian Federation, Moscow, Russia.
I.V. Davydovsky Moscow City Clinical Hospital, Moscow Department of Healthcare, Moscow, Russia.
Front Immunol. 2024 Oct 21;15:1448515. doi: 10.3389/fimmu.2024.1448515. eCollection 2024.
The mechanisms of the SARS-CoV-2-triggered complex alterations in immune cell activation and production of cytokines in lung tissue remain poorly understood, in part because of the limited use of adequate tissue models that simulate the structure and cell composition of the lung . We developed a novel model of SARS-CoV-2 infection of lung explants, that maintains the intact tissue composition and the viral load for up to 7-10 days. Using this model, we studied cytokine production during SARS-CoV-2 infection.
Lung tissue was monitored for viability and cell composition using flow cytometry and histological analysis. SARS-CoV-2 infection was verified immunohistochemically, viral loads in tissue and culture medium were monitored by qPCR. A panel of 41 cytokines was measured in culture medium using xMAP technology.
The explant lung tissue was viable and maintained viral infection that influenced the cytokine production. Elevated concentrations of G-CSF, GM-CSF, GRO-a, IFN-g, IL-6, IL-8, IP-10, MCP-3, MIP-1a, PDGF-AA, and VEGF, and decreased IL-1RA concentration were observed in infected tissue compared to non-infected tissue.
Our results generally reflect the data obtained in COVID-19 patients. GRO-a, IFN-g, IL-6, IL-8, MCP-1, MCP-3, and RANTES correlated with the viral load, forming a distinct pro-inflammatory cluster. Thus, our lung model faithfully reproduces some aspects of cytokine alterations in COVID-19 patients at an early disease stage, making the investigation of SARS-CoV-2 infection mechanisms more accessible and providing a potential platform for antiviral drug testing.
SARS-CoV-2 引发的肺部免疫细胞激活和细胞因子产生的复杂变化的机制仍不清楚，部分原因是有限地使用了合适的组织模型来模拟肺部的结构和细胞组成。我们开发了一种新型的 SARS-CoV-2 感染肺组织外植体模型，该模型可以维持完整的组织组成和病毒载量长达 7-10 天。使用该模型，我们研究了 SARS-CoV-2 感染期间细胞因子的产生。
使用流式细胞术和组织学分析监测肺组织的活力和细胞组成。通过免疫组织化学验证 SARS-CoV-2 感染，通过 qPCR 监测组织和培养基中的病毒载量。使用 xMAP 技术在培养基中测量了 41 种细胞因子。
外植体肺组织具有活力，并维持了影响细胞因子产生的病毒感染。与未感染组织相比，感染组织中 G-CSF、GM-CSF、GRO-a、IFN-g、IL-6、IL-8、IP-10、MCP-3、MIP-1a、PDGF-AA 和 VEGF 的浓度升高，而 IL-1RA 的浓度降低。
我们的结果总体上反映了 COVID-19 患者获得的数据。GRO-a、IFN-g、IL-6、IL-8、MCP-1、MCP-3 和 RANTES 与病毒载量相关，形成了一个独特的促炎簇。因此，我们的肺模型忠实地再现了 COVID-19 患者在疾病早期阶段细胞因子改变的某些方面，使 SARS-CoV-2 感染机制的研究更加容易，并为抗病毒药物测试提供了一个潜在的平台。
