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Changes in the cellular proteins of A549 infected with hepatitis E virus by proteomics analysis.

Shen Q, Pu Y, Fu X, Xie Y, Bian X, Yang S, Yang Y, Cui L, Wang X, Wang H, Zhang W - BMC Vet. Res. (2014)

Bottom Line: Understanding the changes of cellular proteins in these cells exposed to HEV is helpful for elucidating molecular mechanisms associated with function alterations of HEV-infected susceptible cells.Of 2 585-3 152 protein spots visualized on each gel using silver staining, a total of 31 protein spots were found to be differentially expressed in HEV-infected A549 cells compared with mock-infected A549, including 10 significantly up-regulated protein spots and 21 significantly down-regulated protein spots.This work is helpful for investigating the molecular basis associated with the interaction between HEV and the host cells although more efforts should be required to discover the mechanisms.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Medicine, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, Jiangsu, PR China. z0216wen@ujs.edu.cn.

ABSTRACT

Background: Our understanding of Hepatitis E virus (HEV) has changed enormously over the past 30 years, from a waterborne infection causing outbreaks of acute hepatitis in developing countries to an infection of global distribution causing a range of hepatic and extra-hepatic illness. However, the key proteins playing important parts in the virus infection were still unknown. Understanding the changes of cellular proteins in these cells exposed to HEV is helpful for elucidating molecular mechanisms associated with function alterations of HEV-infected susceptible cells. In the present study, a comparative gel-based proteomic analysis was employed to study the changes in cellular proteins of A549 exposed to HEV in vitro to provide novel information for understanding the functional alterations of A549 induced by HEV infection.

Result: Of 2 585-3 152 protein spots visualized on each gel using silver staining, a total of 31 protein spots were found to be differentially expressed in HEV-infected A549 cells compared with mock-infected A549, including 10 significantly up-regulated protein spots and 21 significantly down-regulated protein spots.

Conclusion: Our work is the first time regarding the proteomic analysis on the cellular responses to HEV infection. This work is helpful for investigating the molecular basis associated with the interaction between HEV and the host cells although more efforts should be required to discover the mechanisms.

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Related in: MedlinePlus

Detection of HEV RNA (A), Virus Particles (B) and Antigens (C) in infected cells with RT-PCR, Dot-blot and unlabeled immune Electron Microscopy, respectively. A: 1, infected A549 cells. 2, HEV negative fecal sample inoculated cells. P, HEV positive fecal sample as control; B: F, HEV positive fecal sample. P, HEV inoculated A549 cells. C, HEV negative fecal sample inoculated cells; C, black arrow indicated complete virus particle and white arrow indicated empty virus particle.
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Figure 2: Detection of HEV RNA (A), Virus Particles (B) and Antigens (C) in infected cells with RT-PCR, Dot-blot and unlabeled immune Electron Microscopy, respectively. A: 1, infected A549 cells. 2, HEV negative fecal sample inoculated cells. P, HEV positive fecal sample as control; B: F, HEV positive fecal sample. P, HEV inoculated A549 cells. C, HEV negative fecal sample inoculated cells; C, black arrow indicated complete virus particle and white arrow indicated empty virus particle.

Mentions: Visible CPE was observed at 24 h post-infection, and 90% CPE appeared on day 6 post-infection (Figure 1A-C). Structural changes of infected cells were investigated by TEM. Results showed that the number of lysosome increased, which is an important indicator of apoptosis, and cell membrane damage induced by viral infection (Figure 1D and E). HEV RNA was also detected in the infected cells by RT-PCR method (Figure 2A). Dot-blot results indicated the reactivity of anti-HEV specific antibody with the capsid proteins in the A549 cell (Figure 2B). Figure 2C indicated the unlabeled immune electron micrographs of HEV particles in the infected cells with a diameter about 30 nm. Electron micrograph revealed two morphology forms, one was the empty particle with no significant RNA-like density inside and another was the complete form, which consisted with previous report [22].


Changes in the cellular proteins of A549 infected with hepatitis E virus by proteomics analysis.

Shen Q, Pu Y, Fu X, Xie Y, Bian X, Yang S, Yang Y, Cui L, Wang X, Wang H, Zhang W - BMC Vet. Res. (2014)

Detection of HEV RNA (A), Virus Particles (B) and Antigens (C) in infected cells with RT-PCR, Dot-blot and unlabeled immune Electron Microscopy, respectively. A: 1, infected A549 cells. 2, HEV negative fecal sample inoculated cells. P, HEV positive fecal sample as control; B: F, HEV positive fecal sample. P, HEV inoculated A549 cells. C, HEV negative fecal sample inoculated cells; C, black arrow indicated complete virus particle and white arrow indicated empty virus particle.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4236826&req=5

Figure 2: Detection of HEV RNA (A), Virus Particles (B) and Antigens (C) in infected cells with RT-PCR, Dot-blot and unlabeled immune Electron Microscopy, respectively. A: 1, infected A549 cells. 2, HEV negative fecal sample inoculated cells. P, HEV positive fecal sample as control; B: F, HEV positive fecal sample. P, HEV inoculated A549 cells. C, HEV negative fecal sample inoculated cells; C, black arrow indicated complete virus particle and white arrow indicated empty virus particle.
Mentions: Visible CPE was observed at 24 h post-infection, and 90% CPE appeared on day 6 post-infection (Figure 1A-C). Structural changes of infected cells were investigated by TEM. Results showed that the number of lysosome increased, which is an important indicator of apoptosis, and cell membrane damage induced by viral infection (Figure 1D and E). HEV RNA was also detected in the infected cells by RT-PCR method (Figure 2A). Dot-blot results indicated the reactivity of anti-HEV specific antibody with the capsid proteins in the A549 cell (Figure 2B). Figure 2C indicated the unlabeled immune electron micrographs of HEV particles in the infected cells with a diameter about 30 nm. Electron micrograph revealed two morphology forms, one was the empty particle with no significant RNA-like density inside and another was the complete form, which consisted with previous report [22].

Bottom Line: Understanding the changes of cellular proteins in these cells exposed to HEV is helpful for elucidating molecular mechanisms associated with function alterations of HEV-infected susceptible cells.Of 2 585-3 152 protein spots visualized on each gel using silver staining, a total of 31 protein spots were found to be differentially expressed in HEV-infected A549 cells compared with mock-infected A549, including 10 significantly up-regulated protein spots and 21 significantly down-regulated protein spots.This work is helpful for investigating the molecular basis associated with the interaction between HEV and the host cells although more efforts should be required to discover the mechanisms.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Medicine, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, Jiangsu, PR China. z0216wen@ujs.edu.cn.

ABSTRACT

Background: Our understanding of Hepatitis E virus (HEV) has changed enormously over the past 30 years, from a waterborne infection causing outbreaks of acute hepatitis in developing countries to an infection of global distribution causing a range of hepatic and extra-hepatic illness. However, the key proteins playing important parts in the virus infection were still unknown. Understanding the changes of cellular proteins in these cells exposed to HEV is helpful for elucidating molecular mechanisms associated with function alterations of HEV-infected susceptible cells. In the present study, a comparative gel-based proteomic analysis was employed to study the changes in cellular proteins of A549 exposed to HEV in vitro to provide novel information for understanding the functional alterations of A549 induced by HEV infection.

Result: Of 2 585-3 152 protein spots visualized on each gel using silver staining, a total of 31 protein spots were found to be differentially expressed in HEV-infected A549 cells compared with mock-infected A549, including 10 significantly up-regulated protein spots and 21 significantly down-regulated protein spots.

Conclusion: Our work is the first time regarding the proteomic analysis on the cellular responses to HEV infection. This work is helpful for investigating the molecular basis associated with the interaction between HEV and the host cells although more efforts should be required to discover the mechanisms.

Show MeSH
Related in: MedlinePlus