Limits...
Differential Analysis of the Secretome of WRL68 Cells Infected with the Chikungunya Virus.

Thio CL, Yusof R, Ashrafzadeh A, Bahari S, Abdul-Rahman PS, Karsani SA - PLoS ONE (2015)

Bottom Line: These results provide an initial view of CHIKV may affect the secretome of infected cells during early infection.The results presented here will compliment earlier results from the study of late host response.However, functional characterization will be necessary to further enhance our understanding of the roles played by these proteins in the early stages of CHIKV infection in humans.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia; Drug Design and Development Research Group (DDDRG), University of Malaya, 50603, Kuala Lumpur, Malaysia.

ABSTRACT
The Chikungunya virus (CHIKV) is an arthropod borne virus. In the last 50 years, it has been the cause of numerous outbreaks in tropical and temperate regions, worldwide. There is limited understanding regarding the underlying molecular mechanisms involved in CHIKV replication and how the virus interacts with its host. In the present study, comparative proteomics was used to identify secreted host proteins that changed in abundance in response to early CHIKV infection. Two-dimensional gel electrophoresis was used to analyse and compare the secretome profiles of WRL-68 cells infected with CHIKV against mock control WRL-68 cells. The analysis identified 25 regulated proteins in CHIKV infected cells. STRING network analysis was then used to predict biological processes that may be affected by these proteins. The processes predicted to be affected include signal transduction, cellular component and extracellular matrix (ECM) organization, regulation of cytokine stimulus and immune response. These results provide an initial view of CHIKV may affect the secretome of infected cells during early infection. The results presented here will compliment earlier results from the study of late host response. However, functional characterization will be necessary to further enhance our understanding of the roles played by these proteins in the early stages of CHIKV infection in humans.

No MeSH data available.


Related in: MedlinePlus

Relative cell viability (A) and percentage of cell lysis (B) of WRL-68 cells grown in DMEM growth medium (10% FBS) and serum-free medium for 24 hours.A: Cell viability was normalised to 100%, and the relative viability of serum-starved cells was determined. At 24 hours incubation, the absence of serum did not significantly reduce cell viability as percentage of viability was above 98%. Data presented are representative of three independent experiments and the error bars represent standard deviation. B: Percentage of cell lysis for digitonin-lysed cells (positive control) was normalised to 100%. The absence of serum in the culture medium was found to have no significant effect on cellular disruption. Similarly, WRL-68 cells infected with CHIKV at the MOI of 5.0 showed similar percentage as the mock control cells. Data presented are representative of three independent experiments and the error bars represent standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4470940&req=5

pone.0129033.g001: Relative cell viability (A) and percentage of cell lysis (B) of WRL-68 cells grown in DMEM growth medium (10% FBS) and serum-free medium for 24 hours.A: Cell viability was normalised to 100%, and the relative viability of serum-starved cells was determined. At 24 hours incubation, the absence of serum did not significantly reduce cell viability as percentage of viability was above 98%. Data presented are representative of three independent experiments and the error bars represent standard deviation. B: Percentage of cell lysis for digitonin-lysed cells (positive control) was normalised to 100%. The absence of serum in the culture medium was found to have no significant effect on cellular disruption. Similarly, WRL-68 cells infected with CHIKV at the MOI of 5.0 showed similar percentage as the mock control cells. Data presented are representative of three independent experiments and the error bars represent standard deviation.

Mentions: MTS was used to investigate the viability of serum-starved WRL-68 cells. Fig 1A shows the relative viability of cells grown in normal DMEM medium (10% FBS) and serum-free medium where no significant difference in cell viability was observed.


Differential Analysis of the Secretome of WRL68 Cells Infected with the Chikungunya Virus.

Thio CL, Yusof R, Ashrafzadeh A, Bahari S, Abdul-Rahman PS, Karsani SA - PLoS ONE (2015)

Relative cell viability (A) and percentage of cell lysis (B) of WRL-68 cells grown in DMEM growth medium (10% FBS) and serum-free medium for 24 hours.A: Cell viability was normalised to 100%, and the relative viability of serum-starved cells was determined. At 24 hours incubation, the absence of serum did not significantly reduce cell viability as percentage of viability was above 98%. Data presented are representative of three independent experiments and the error bars represent standard deviation. B: Percentage of cell lysis for digitonin-lysed cells (positive control) was normalised to 100%. The absence of serum in the culture medium was found to have no significant effect on cellular disruption. Similarly, WRL-68 cells infected with CHIKV at the MOI of 5.0 showed similar percentage as the mock control cells. Data presented are representative of three independent experiments and the error bars represent standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4470940&req=5

pone.0129033.g001: Relative cell viability (A) and percentage of cell lysis (B) of WRL-68 cells grown in DMEM growth medium (10% FBS) and serum-free medium for 24 hours.A: Cell viability was normalised to 100%, and the relative viability of serum-starved cells was determined. At 24 hours incubation, the absence of serum did not significantly reduce cell viability as percentage of viability was above 98%. Data presented are representative of three independent experiments and the error bars represent standard deviation. B: Percentage of cell lysis for digitonin-lysed cells (positive control) was normalised to 100%. The absence of serum in the culture medium was found to have no significant effect on cellular disruption. Similarly, WRL-68 cells infected with CHIKV at the MOI of 5.0 showed similar percentage as the mock control cells. Data presented are representative of three independent experiments and the error bars represent standard deviation.
Mentions: MTS was used to investigate the viability of serum-starved WRL-68 cells. Fig 1A shows the relative viability of cells grown in normal DMEM medium (10% FBS) and serum-free medium where no significant difference in cell viability was observed.

Bottom Line: These results provide an initial view of CHIKV may affect the secretome of infected cells during early infection.The results presented here will compliment earlier results from the study of late host response.However, functional characterization will be necessary to further enhance our understanding of the roles played by these proteins in the early stages of CHIKV infection in humans.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia; Drug Design and Development Research Group (DDDRG), University of Malaya, 50603, Kuala Lumpur, Malaysia.

ABSTRACT
The Chikungunya virus (CHIKV) is an arthropod borne virus. In the last 50 years, it has been the cause of numerous outbreaks in tropical and temperate regions, worldwide. There is limited understanding regarding the underlying molecular mechanisms involved in CHIKV replication and how the virus interacts with its host. In the present study, comparative proteomics was used to identify secreted host proteins that changed in abundance in response to early CHIKV infection. Two-dimensional gel electrophoresis was used to analyse and compare the secretome profiles of WRL-68 cells infected with CHIKV against mock control WRL-68 cells. The analysis identified 25 regulated proteins in CHIKV infected cells. STRING network analysis was then used to predict biological processes that may be affected by these proteins. The processes predicted to be affected include signal transduction, cellular component and extracellular matrix (ECM) organization, regulation of cytokine stimulus and immune response. These results provide an initial view of CHIKV may affect the secretome of infected cells during early infection. The results presented here will compliment earlier results from the study of late host response. However, functional characterization will be necessary to further enhance our understanding of the roles played by these proteins in the early stages of CHIKV infection in humans.

No MeSH data available.


Related in: MedlinePlus