Objective

High mobility group box-1 (HMGB1) has been reported to be involved in influenza A virus-induced acute respiratory distress syndrome (ARDS). We studied the efficacy of an anti-HMGB1 mAb using an in vitro model of TNF-α stimulation or influenza A virus infection in human pulmonary microvascular endothelial cells (HMVECs).

Methods

Vascular permeability of HMVECs was quantified using the Boyden chamber assay under tumor necrosis factor-α (TNF-α) stimulation or influenza A virus infection in the presence of anti-HMGB1 mAb or control mAb. The intracellular localization of HMGB1 was assessed by immunostaining. Extracellular cytokine concentrations and intracellular viral mRNA expression were quantified by the enzyme-linked immunosorbent assay and quantitative reverse transcription PCR, respectively.

Results

Vascular permeability was increased by TNF-α stimulation or influenza A infection; HMVECs became elongated and the intercellular gaps were extended. Anti-HMGB1 mAb suppressed both the increase in permeability and the cell morphology changes. Translocation of HMGB1 to the cytoplasm was observed in the non-infected cells. Although anti-HMGB1 mAb did not suppress viral replication, it did suppress cytokine production in HMVECs.

Conclusion

Anti-HMGB1 mAb might be an effective therapy for severe influenza ARDS.

中文翻译：

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抗高迁移率组框 1 单克隆抗体抑制感染甲型流感病毒的人肺内皮细胞的高通透性和细胞因子产生

客观的

据报道，高迁移率 group box-1 (HMGB1) 与甲型流感病毒引起的急性呼吸窘迫综合征 (ARDS) 相关。我们使用人肺微血管内皮细胞 (HMVEC) 中 TNF-α 刺激或甲型流感病毒感染的体外模型研究了抗 HMGB1 mAb 的功效。

方法

在抗 HMGB1 mAb 或对照 mAb 存在下，在肿瘤坏死因子-α (TNF-α) 刺激或甲型流感病毒感染下，使用 Boyden 室测定法量化 HMVEC 的血管通透性。通过免疫染色评估HMGB1的细胞内定位。细胞外细胞因子浓度和细胞内病毒mRNA表达分别通过酶联免疫吸附测定和定量逆转录PCR定量。

结果

血管通透性因 TNF-α 刺激或甲型流感感染而增加；HMVECs 被拉长并且细胞间的间隙被扩展。抗 HMGB1 mAb 抑制了渗透性的增加和细胞形态的变化。在未感染的细胞中观察到 HMGB1 易位至细胞质。尽管抗 HMGB1 mAb 不抑制病毒复制，但它确实抑制了 HMVEC 中细胞因子的产生。

结论

抗 HMGB1 mAb 可能是治疗严重流感 ARDS 的有效方法。