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The role of SARS-CoV-2 nucleocapsid protein in effecting immune cells and insights on its molecular mechanisms

ABSTRACT: The study aims were to explore the immune regulatory function of SARA-CoV-2 Nucleocapsid (N protein) and the related mechanisms. In a series of protein activity experiments, the SARA-CoV-2 Nucleocapsid showed the proliferation effect on three immune cell lines (Raw264.7, Jurkat and Raji) in a dose-dependent manner. N protein(10μg/mL) could significantly change cell cycle progressing of these three immune cell lines and could promote Raw264.7 cells quickly entering G2/M phase from the S phase to achieve rapid growth. Additionally, N protein((10μg/mL) also could stimulate Raw264.7 cells to secrete a large number of proinflammatory factors such as TNFα, IL6 and IL10. The RNA sequencing analysis indicated that N protein changed the expression of some certain genes significantly in immune related function and four important signaling pathways including JAK-STAT signaling pathway, TNF signaling pathway, NF-kappa B signaling pathway and MAPK signaling pathway, which suggested that N protein could not only regulate the expression of genes involved in the process of resisting virus infection in macrophages of immune system, but also change the cellular signal processing.

ORGANISM(S): Mus musculus

PROVIDER: GSE236800 | GEO | 2023/07/12

REPOSITORIES: GEO

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Project description:Single-walled carbon nanotubes (SWCNTs) are being explored and used for a wide range of applications in industrial and medical sectors, and the increasing exposure of SWCNTs necessitate the studies of their potential environmental and health effects. Considerable efforts have been made to improve the dispersion of SWCNTs by chemical modifications. The present study was designed to determine if the acid functionalization of SWCNTs enhanced the toxicity and more efforts were made to understand the molecular mechanisms. RAW264.7 cells were exposed to 0-100μg/mL of SWCNTs and AF-SWCNTs for 24 hours and cell proliferation, viability, and gene expression profiles were assessed and compared. Results showed that the magnitude of AF-SWCNT-induced cell proliferation inhibition was higher than that of SWCNTs. 10μg/mL of AF-SWCNTs was determined as non-toxic to cells by viability analysis. AF-SWCNTs could enter and aggregate in cell cytoplasm and nuclear areas. Microarray, real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assays (ELISA) demonstrated that AF-SWCNTs (1, 10, and 50μg/mL) altered gene and protein expression patterns. The most differentially expressed genes were related to pro-inflammatory cytokines and chemokines (such as CCL-3 and CCL-4), apoptosis (e.g., Caspase-1, -4, and Dffa), protective response (such as Hmox-1and GSTM3), and cell surface molecules such as ICAM-1, Itgb2, indicating a possible involvement of SWCNT in inflammation, anti-apoptotic and DNA fragmentation, carcinogenesis and biased T cell polarization. Furthermore, a substantial amount of down-regulated genes were found related to ribosomal assembly and mitochondrial respiration chain, implicating a possible mechanism of SWCNT-induced oxidative stress and gene expression inhibition. RAW264.7 gene expression after exposure to 1-50ug/ml AF-SWCNT-24 hours. 0ug/ml of AF-SWCNT was used as control sample. The data were obtained from three biological replicates.

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The role of SARS-CoV-2 nucleocapsid protein in effecting immune cells and insights on its molecular mechanisms

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