Department of Microbiology and Molecular Biology, College of Life Sciences, Brigham Young University, Provo, Utah, United States of America.
Department of Statistics, College of Physical and Mathematical Sciences, Brigham Young University, Provo, Utah, United States of America.
PLoS One. 2020 Mar 12;15(3):e0230328. doi: 10.1371/journal.pone.0230328. eCollection 2020.
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes rash, fever and severe polyarthritis that can last for years in humans. Murine models display inflammation and macrophage infiltration only in the adjacent tissues at the site of inoculation, showing no signs of systemic polyarthritis. Monocyte-derived macrophages are one cell type suspected to contribute to a systemic CHIKV infection. The purpose of this study was to analyze differences in CHIKV infection in two different cell lines, human U937 and murine RAW264.7 monocyte derived macrophages. PMA-differentiated U937 and RAW264.7 macrophages were infected with CHIKV, and infectious virus production was measured by plaque assay and by reverse transcriptase quantitative PCR at various time points. Secreted cytokines in the supernatants were measured using cytometric bead arrays. Cytokine mRNA levels were also measured to supplement expression data. Here we show that CHIKV replicates more efficiently in human macrophages compared to murine macrophages. In addition, infected human macrophages produced around 10-fold higher levels of infectious virus when compared to murine macrophages. Cytokine induction by CHIKV infection differed between human and murine macrophages; IL-1, IL-6, IFN-γ, and TNF were significantly upregulated in human macrophages. This evidence suggests that CHIKV replicates more efficiently and induces a much greater pro-inflammatory cytokine profile in human macrophages, when compared to murine macrophages. This may shed light on the critical role that macrophages play in the CHIKV inflammatory response.
基孔肯雅病毒(CHIKV)是一种通过蚊子传播的甲病毒,可引起皮疹、发热和严重的多发性关节炎,这些症状在人类中可能持续数年。在鼠模型中,仅在接种部位的相邻组织中显示炎症和巨噬细胞浸润,没有全身多发性关节炎的迹象。单核细胞衍生的巨噬细胞是一种被怀疑有助于全身性 CHIKV 感染的细胞类型。本研究的目的是分析两种不同细胞系,人 U937 和鼠 RAW264.7 单核细胞衍生巨噬细胞中 CHIKV 感染的差异。用佛波醇酯(PMA)分化 U937 和 RAW264.7 巨噬细胞,然后用空斑试验和逆转录定量 PCR 测定不同时间点的感染性病毒产量。使用细胞因子珠阵列测量上清液中分泌的细胞因子。还测量细胞因子 mRNA 水平以补充表达数据。我们在这里表明,与鼠巨噬细胞相比,CHIKV 在人巨噬细胞中复制效率更高。此外,与鼠巨噬细胞相比,感染的人巨噬细胞产生的感染性病毒水平高约 10 倍。CHIKV 感染引起的细胞因子诱导在人巨噬细胞和鼠巨噬细胞之间存在差异;人巨噬细胞中 IL-1、IL-6、IFN-γ 和 TNF 显著上调。这一证据表明,与鼠巨噬细胞相比,CHIKV 在人巨噬细胞中复制效率更高,并诱导更强的促炎细胞因子谱。这可能揭示了巨噬细胞在 CHIKV 炎症反应中发挥的关键作用。