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Establishment of a Novel Primary Human Skeletal Myoblast Cellular Model for Chikungunya Virus Infection and Pathogenesis.

Hussain KM, Lee RC, Ng MM, Chu JJ - Sci Rep (2016)

Bottom Line: Chikungunya virus (CHIKV) is a re-emerging arbovirus known to cause chronic myalgia and arthralgia and is now considered endemic in countries across Asia and Africa.Infection of HSMM cells with CHIKV resulted in altered expressions of host genes involved in skeletal- and muscular-associated disorders, innate immune responses, cellular growth and death, host metabolism and virus replication.Together, this study has shown the establishment of a clinically relevant primary human cell model that paves the way for the further analysis of host factors and their involvement in the various stages of CHIKV replication cycle and viral pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular RNA Virology and Antiviral Strategies, Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore.

ABSTRACT
Chikungunya virus (CHIKV) is a re-emerging arbovirus known to cause chronic myalgia and arthralgia and is now considered endemic in countries across Asia and Africa. The tissue tropism of CHIKV infection in humans remains, however, ill-defined. Due to the fact that myositis is commonly observed in most patients infected with CHIKV, we sought to develop a clinically relevant cellular model to better understand the pathogenesis of CHIKV infection. In this study, primary human skeletal muscle myoblasts (HSMM) were established as a novel human primary cell line that is highly permissive to CHIKV infection, with maximal amounts of infectious virions observed at 16 hours post infection. Genome-wide microarray profiling analyses were subsequently performed to identify and map genes that are differentially expressed upon CHIKV infection. Infection of HSMM cells with CHIKV resulted in altered expressions of host genes involved in skeletal- and muscular-associated disorders, innate immune responses, cellular growth and death, host metabolism and virus replication. Together, this study has shown the establishment of a clinically relevant primary human cell model that paves the way for the further analysis of host factors and their involvement in the various stages of CHIKV replication cycle and viral pathogenesis.

No MeSH data available.


Related in: MedlinePlus

Immunofluorescence assay depicting CHIKV replication in HSMM cells (a) Production of CHIKV envelope protein (FITC) could be observed by 12 h.p.i., with almost complete infection of all HSMM cells observed by 72 h.p.i. (100x magnification). (b) At higher magnification, CHIKV viral antigens (arrows) are observed at the perinuclear region as early as 8 h.p.i. and subsequently localized to the cellular periphery by 24 h.p.i. (1000x magnification; arrows). Cell nuclei are counterstained with DAPI.
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f2: Immunofluorescence assay depicting CHIKV replication in HSMM cells (a) Production of CHIKV envelope protein (FITC) could be observed by 12 h.p.i., with almost complete infection of all HSMM cells observed by 72 h.p.i. (100x magnification). (b) At higher magnification, CHIKV viral antigens (arrows) are observed at the perinuclear region as early as 8 h.p.i. and subsequently localized to the cellular periphery by 24 h.p.i. (1000x magnification; arrows). Cell nuclei are counterstained with DAPI.

Mentions: CHIKV (strain Singapore 072008) replication in HSMM cells was also analysed by performing an immunofluorescence assay. The production of the FITC-stained CHIKV envelope protein could be observed by 12 h.p.i. (Fig. 2a), with almost-complete infection rate by 72 h.p.i. In the early stages of infection, the viral E2 antigen could be observed within the perinuclear region of the cell (Fig. 2b). As infection progressed, the viral proteins (arrows) were observed to be accumulating within the cytoplasm and towards the plasma membrane. It is known that the viral envelope proteins are synthesized at the rough ER and transported to the plasma membrane for virus assembly and budding to release the infectious virus particles.


Establishment of a Novel Primary Human Skeletal Myoblast Cellular Model for Chikungunya Virus Infection and Pathogenesis.

Hussain KM, Lee RC, Ng MM, Chu JJ - Sci Rep (2016)

Immunofluorescence assay depicting CHIKV replication in HSMM cells (a) Production of CHIKV envelope protein (FITC) could be observed by 12 h.p.i., with almost complete infection of all HSMM cells observed by 72 h.p.i. (100x magnification). (b) At higher magnification, CHIKV viral antigens (arrows) are observed at the perinuclear region as early as 8 h.p.i. and subsequently localized to the cellular periphery by 24 h.p.i. (1000x magnification; arrows). Cell nuclei are counterstained with DAPI.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4759813&req=5

f2: Immunofluorescence assay depicting CHIKV replication in HSMM cells (a) Production of CHIKV envelope protein (FITC) could be observed by 12 h.p.i., with almost complete infection of all HSMM cells observed by 72 h.p.i. (100x magnification). (b) At higher magnification, CHIKV viral antigens (arrows) are observed at the perinuclear region as early as 8 h.p.i. and subsequently localized to the cellular periphery by 24 h.p.i. (1000x magnification; arrows). Cell nuclei are counterstained with DAPI.
Mentions: CHIKV (strain Singapore 072008) replication in HSMM cells was also analysed by performing an immunofluorescence assay. The production of the FITC-stained CHIKV envelope protein could be observed by 12 h.p.i. (Fig. 2a), with almost-complete infection rate by 72 h.p.i. In the early stages of infection, the viral E2 antigen could be observed within the perinuclear region of the cell (Fig. 2b). As infection progressed, the viral proteins (arrows) were observed to be accumulating within the cytoplasm and towards the plasma membrane. It is known that the viral envelope proteins are synthesized at the rough ER and transported to the plasma membrane for virus assembly and budding to release the infectious virus particles.

Bottom Line: Chikungunya virus (CHIKV) is a re-emerging arbovirus known to cause chronic myalgia and arthralgia and is now considered endemic in countries across Asia and Africa.Infection of HSMM cells with CHIKV resulted in altered expressions of host genes involved in skeletal- and muscular-associated disorders, innate immune responses, cellular growth and death, host metabolism and virus replication.Together, this study has shown the establishment of a clinically relevant primary human cell model that paves the way for the further analysis of host factors and their involvement in the various stages of CHIKV replication cycle and viral pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular RNA Virology and Antiviral Strategies, Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore.

ABSTRACT
Chikungunya virus (CHIKV) is a re-emerging arbovirus known to cause chronic myalgia and arthralgia and is now considered endemic in countries across Asia and Africa. The tissue tropism of CHIKV infection in humans remains, however, ill-defined. Due to the fact that myositis is commonly observed in most patients infected with CHIKV, we sought to develop a clinically relevant cellular model to better understand the pathogenesis of CHIKV infection. In this study, primary human skeletal muscle myoblasts (HSMM) were established as a novel human primary cell line that is highly permissive to CHIKV infection, with maximal amounts of infectious virions observed at 16 hours post infection. Genome-wide microarray profiling analyses were subsequently performed to identify and map genes that are differentially expressed upon CHIKV infection. Infection of HSMM cells with CHIKV resulted in altered expressions of host genes involved in skeletal- and muscular-associated disorders, innate immune responses, cellular growth and death, host metabolism and virus replication. Together, this study has shown the establishment of a clinically relevant primary human cell model that paves the way for the further analysis of host factors and their involvement in the various stages of CHIKV replication cycle and viral pathogenesis.

No MeSH data available.


Related in: MedlinePlus