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To investigate the effect and mechanism of HBV-stimulated HTR8-S/Vneo mitochondrial antiviral signaling protein (MAVS) signaling pathway on the immune response of peripheral blood mononuclear cell (PBMC) to hepatitis B vaccine.
The HTR8-S/Vneo were co-incubated with serum of HBV-DNA positive patients. MAVS siRNA transfection was hereby applied for silencing of MAVS and MAVS plasmid transfection was applied for overexpression of MAVS in HTR8-S/Vneo. We then cocultured them with human immune cells assembly PBMC and subsequently stimulated with hepatitis B vaccine. The mRNA and protein expression levels of MAVS, NF-κB, pNF-κB, IRF3 and pIRF3 in HTR8-S/Vneo were detected by reverse transcription-quantitative real-time PCR (RT-qPCR) and flow cytometry (FCM). The levels of IL-2, IL-6, IFN-β and IL-17 were detected by enzyme-linked immunosorbent assay (ELISA), and the proportion of CD4+T cell were detected by flow cytometry. The data were analyzed for t test.
HBV down-regulated the mRNA expression levels of MAVS, NF-κB, IRF3 (t=6.66, P=0.003; t=14.18, P<0.001; t=3.70, P=0.021) and the protein expression levels of MAVS, NF-κB, pNF-κB (t=3.42, P=0.042; t=4.23, P=0.013; t=4.86, P=0.008), and inhibited the production of IFN-β (t=9.83, P=0.010). Silencing MAVS down-regulated the mRNA and protein expression of MAVS (t=31.20, P<0.001; t=6.53, P=0.023), inhibited the expression of IFN-β (t=5.46, P=0.032), and reduced the proportion of CD4+T cells in PBMC (t=9.07, P=0.001). Overexpression of MAVS induced IFN-β production (t=-34.26, P<0.001; t=-5.11, P=0.036), but the proportion of CD4+T cells did not show significant effect (t=-0.73, P=0.506).
HBV can inhibit the MAVS signaling pathway in HTR8-S/Vneo. Overexpression of MAVS could promote the immune response to hepatitis B vaccine in PBMC.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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