Purpose

To investigate the transcriptome of human bronchial epithelial cells (HBEC) in response to serum from patients with different degrees of inflammation.

Methods

Serum from 19 COVID-19 patients obtained from the Hannover Unified Biobank was used. At the time of sampling, 5 patients had a WHO Clinical Progression Scale (WHO-CPS) score of 9 (severe illness). The remaining 14 patients had a WHO-CPS of below 9 (range 1–7), and lower illness. Multiplex immunoassay was used to assess serum inflammatory markers. The culture medium of HBEC was supplemented with 2% of the patient’s serum, and the cells were cultured at 37 °C, 5% CO₂ for 18 h. Subsequently, cellular RNA was used for RNA-Seq.

Results

Patients with scores below 9 had significantly lower albumin and serum levels of E-selectin, IL-8, and MCP-1 than patients with scores of 9. Principal component analysis based on 500 “core genes” of RNA-seq segregated cells into two subsets: exposed to serum from 4 (I) and 15 (II) patients. Cells from a subset (I) treated with serum from 4 patients with a score of 9 showed 5566 differentially expressed genes of which 2793 were up- and 2773 downregulated in comparison with cells of subset II treated with serum from 14 patients with scores between 1 and 7 and one with score = 9. In subset I cells, a higher expression of TLR4 and CXCL8 but a lower CDH1, ACE2, and HMOX1, and greater effects on genes involved in metabolic regulation, cytoskeletal organization, and kinase activity pathways were observed.

Conclusion

This simple model could be useful to characterize patient serum and epithelial cell properties.

中文翻译：

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人支气管上皮细胞转录组对不同炎症状态患者血清反应的变化

目的

研究人支气管上皮细胞（HBEC）对不同程度炎症患者血清的转录组反应。

方法

使用从汉诺威统一生物库获得的 19 名 COVID-19 患者的血清。采样时，5 名患者的世界卫生组织临床进展量表 (WHO-CPS) 评分为 9 分（病情严重）。其余 14 名患者的 WHO-CPS 低于 9（范围 1-7），且病情较低。使用多重免疫测定来评估血清炎症标志物。HBEC培养基中添加2%患者血清，37℃、5%CO ₂培养细胞18 h。随后，细胞RNA被用于RNA-Seq。

结果

得分低于 9 的患者的白蛋白和血清 E-选择素、IL-8 和 MCP-1 水平显着低于得分为 9 的患者。基于 RNA-seq 将细胞分为两部分的 500 个“核心基因”的主成分分析亚组：暴露于 4 (I) 和 15 (II) 名患者的血清。用来自 4 名得分为 9 的患者的血清处理的子集 (I) 的细胞显示出 5566 个差异表达基因，其中与用来自 14 名得分在 1 至 1 之间的患者的血清处理的子集 II 的细胞相比，其中 2793 个上调和 2773 个下调。 7 分和评分 = 9 分的细胞中。在 I 子集细胞中，观察到TLR4和CXCL8表达较高，但CDH1 、 ACE2和HMOX1表达较低，并且对参与代谢调节、细胞骨架组织和激酶活性途径的基因影响较大。

结论

这个简单的模型可用于表征患者血清和上皮细胞的特性。