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MicroRNA-19b-3p Modulates Japanese Encephalitis Virus-Mediated Inflammation via Targeting RNF11.

Ashraf U, Zhu B, Ye J, Wan S, Nie Y, Chen Z, Cui M, Wang C, Duan X, Zhang H, Chen H, Cao S - J. Virol. (2016)

Bottom Line: Japanese encephalitis virus (JEV) can invade the central nervous system and consequently induce neuroinflammation, which is characterized by profound neuronal cell damage accompanied by astrogliosis and microgliosis.The pathological features of JEV-induced encephalitis are inflammatory reactions and neurological diseases resulting from glia activation.The present study reveals that miR-19b-3p targets ring finger protein 11 in glia and promotes inflammatory cytokine production by enhancing nuclear factor kappa B activity in these cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.

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miR-19b-3p expression is upregulated after JEV infection. (A and B) U251 cells were infected with JEV at an MOI of 5 for the indicated times (A) or at the indicated MOIs for 36 h (B). (C) U251 cells were incubated with UV-inactivated JEV at an MOI of 5 for 36 h. (D and E) BV2 cells were infected with JEV at an MOI of 5 for the indicated times (D) or at the indicated MOIs for 36 h (E). The levels of miR-19b-3p were detected by quantitative real-time PCR (upper panels). Western blotting was performed to examine the expression of JEV NS5 protein (lower panels). GAPDH expression was verified as a loading control. All data are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Con, control.
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Figure 1: miR-19b-3p expression is upregulated after JEV infection. (A and B) U251 cells were infected with JEV at an MOI of 5 for the indicated times (A) or at the indicated MOIs for 36 h (B). (C) U251 cells were incubated with UV-inactivated JEV at an MOI of 5 for 36 h. (D and E) BV2 cells were infected with JEV at an MOI of 5 for the indicated times (D) or at the indicated MOIs for 36 h (E). The levels of miR-19b-3p were detected by quantitative real-time PCR (upper panels). Western blotting was performed to examine the expression of JEV NS5 protein (lower panels). GAPDH expression was verified as a loading control. All data are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Con, control.

Mentions: To interrogate the aftermath of JEV infection on the miRNA profile, miRNA deep-sequencing analysis of JEV-infected U251 cells was performed. Our sequencing data indicated that a group of miRNAs were differentially regulated upon JEV infection. Of these miRNAs, miR-19b-3p was found to be statistically well upregulated upon viral infection, and it was expressed at high levels in the cells (unpublished data). Because miR-19b-3p was found to be engaged in the cytokine- and chemokine-mediated inflammatory pathway (47), it was selected for further characterization during JEV infection. The corroboration of miR-19b-3p expression patterns in JEV-infected U251 cells was scrutinized by quantitative real-time PCR. The results revealed that miR-19b-3p was significantly upregulated in a time-dependent (Fig. 1A) and dose-dependent (Fig. 1B) manner. In contrast to these findings, UV-irradiated inactivated JEV infection failed to induce miR-19b-3p upregulation in U251 cells (Fig. 1C), suggesting possible involvement of miR-19b-3p in a JEV-induced inflammatory response in brain astrocytes. We also investigated the expression of miR-19b-3p in JEV-infected microglial BV2 cells (Fig. 1D and E). The results were similar to those observed in JEV-infected U251 cells. These results strongly demonstrate that miR-19b-3p expression is upregulated after JEV infection.


MicroRNA-19b-3p Modulates Japanese Encephalitis Virus-Mediated Inflammation via Targeting RNF11.

Ashraf U, Zhu B, Ye J, Wan S, Nie Y, Chen Z, Cui M, Wang C, Duan X, Zhang H, Chen H, Cao S - J. Virol. (2016)

miR-19b-3p expression is upregulated after JEV infection. (A and B) U251 cells were infected with JEV at an MOI of 5 for the indicated times (A) or at the indicated MOIs for 36 h (B). (C) U251 cells were incubated with UV-inactivated JEV at an MOI of 5 for 36 h. (D and E) BV2 cells were infected with JEV at an MOI of 5 for the indicated times (D) or at the indicated MOIs for 36 h (E). The levels of miR-19b-3p were detected by quantitative real-time PCR (upper panels). Western blotting was performed to examine the expression of JEV NS5 protein (lower panels). GAPDH expression was verified as a loading control. All data are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Con, control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4836334&req=5

Figure 1: miR-19b-3p expression is upregulated after JEV infection. (A and B) U251 cells were infected with JEV at an MOI of 5 for the indicated times (A) or at the indicated MOIs for 36 h (B). (C) U251 cells were incubated with UV-inactivated JEV at an MOI of 5 for 36 h. (D and E) BV2 cells were infected with JEV at an MOI of 5 for the indicated times (D) or at the indicated MOIs for 36 h (E). The levels of miR-19b-3p were detected by quantitative real-time PCR (upper panels). Western blotting was performed to examine the expression of JEV NS5 protein (lower panels). GAPDH expression was verified as a loading control. All data are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Con, control.
Mentions: To interrogate the aftermath of JEV infection on the miRNA profile, miRNA deep-sequencing analysis of JEV-infected U251 cells was performed. Our sequencing data indicated that a group of miRNAs were differentially regulated upon JEV infection. Of these miRNAs, miR-19b-3p was found to be statistically well upregulated upon viral infection, and it was expressed at high levels in the cells (unpublished data). Because miR-19b-3p was found to be engaged in the cytokine- and chemokine-mediated inflammatory pathway (47), it was selected for further characterization during JEV infection. The corroboration of miR-19b-3p expression patterns in JEV-infected U251 cells was scrutinized by quantitative real-time PCR. The results revealed that miR-19b-3p was significantly upregulated in a time-dependent (Fig. 1A) and dose-dependent (Fig. 1B) manner. In contrast to these findings, UV-irradiated inactivated JEV infection failed to induce miR-19b-3p upregulation in U251 cells (Fig. 1C), suggesting possible involvement of miR-19b-3p in a JEV-induced inflammatory response in brain astrocytes. We also investigated the expression of miR-19b-3p in JEV-infected microglial BV2 cells (Fig. 1D and E). The results were similar to those observed in JEV-infected U251 cells. These results strongly demonstrate that miR-19b-3p expression is upregulated after JEV infection.

Bottom Line: Japanese encephalitis virus (JEV) can invade the central nervous system and consequently induce neuroinflammation, which is characterized by profound neuronal cell damage accompanied by astrogliosis and microgliosis.The pathological features of JEV-induced encephalitis are inflammatory reactions and neurological diseases resulting from glia activation.The present study reveals that miR-19b-3p targets ring finger protein 11 in glia and promotes inflammatory cytokine production by enhancing nuclear factor kappa B activity in these cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.

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Related in: MedlinePlus