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Characterization of reemerging chikungunya virus.

Sourisseau M, Schilte C, Casartelli N, Trouillet C, Guivel-Benhassine F, Rudnicka D, Sol-Foulon N, Le Roux K, Prevost MC, Fsihi H, Frenkiel MP, Blanchet F, Afonso PV, Ceccaldi PE, Ozden S, Gessain A, Schuffenecker I, Verhasselt B, Zamborlini A, Saïb A, Rey FA, Arenzana-Seisdedos F, Desprès P, Michault A, Albert ML, Schwartz O - PLoS Pathog. (2007)

Bottom Line: CHIKV replication was cytopathic and associated with an induction of apoptosis in infected cells.CHIKV was highly sensitive to the antiviral activity of type I and II interferons.These results provide a general insight into the interaction between CHIKV and its mammalian host.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Unité Virus et Immunité, Institut Pasteur, Paris, France.

ABSTRACT
An unprecedented epidemic of chikungunya virus (CHIKV) infection recently started in countries of the Indian Ocean area, causing an acute and painful syndrome with strong fever, asthenia, skin rash, polyarthritis, and lethal cases of encephalitis. The basis for chikungunya disease and the tropism of CHIKV remain unknown. Here, we describe the replication characteristics of recent clinical CHIKV strains. Human epithelial and endothelial cells, primary fibroblasts and, to a lesser extent, monocyte-derived macrophages, were susceptible to infection and allowed viral production. In contrast, CHIKV did not replicate in lymphoid and monocytoid cell lines, primary lymphocytes and monocytes, or monocyte-derived dendritic cells. CHIKV replication was cytopathic and associated with an induction of apoptosis in infected cells. Chloroquine, bafilomycin-A1, and short hairpin RNAs against dynamin-2 inhibited viral production, indicating that viral entry occurs through pH-dependent endocytosis. CHIKV was highly sensitive to the antiviral activity of type I and II interferons. These results provide a general insight into the interaction between CHIKV and its mammalian host.

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Related in: MedlinePlus

Assembly and Release of CHIKV(A) Western blot analysis of CHIKV proteins. HeLa cells were exposed to the indicated CHIKV strains (moi 10). Cell lysates and pelleted supernatants were analyzed by Western blot 24 h pi, with a mix of anti-capsid mAb and anti-CHIKV antibodies. The predicted viral proteins are indicated on the right.(B) Gallery of electron micrographs of CHIKV-infected HeLa cells. Cells were analyzed at 36 h pi. CHIKV mostly buds at the plasma membrane of HeLa cells. Viral particles were not detected in noninfected cells (not shown). Bars represent 1 μm in the upper panels and 100 nm in the lower panels.NI, noninfected cells.
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ppat-0030089-g003: Assembly and Release of CHIKV(A) Western blot analysis of CHIKV proteins. HeLa cells were exposed to the indicated CHIKV strains (moi 10). Cell lysates and pelleted supernatants were analyzed by Western blot 24 h pi, with a mix of anti-capsid mAb and anti-CHIKV antibodies. The predicted viral proteins are indicated on the right.(B) Gallery of electron micrographs of CHIKV-infected HeLa cells. Cells were analyzed at 36 h pi. CHIKV mostly buds at the plasma membrane of HeLa cells. Viral particles were not detected in noninfected cells (not shown). Bars represent 1 μm in the upper panels and 100 nm in the lower panels.NI, noninfected cells.

Mentions: We also characterized CHIKV by Western blot analysis. Various viral proteins were detected in lysates of CHIKV-infected HeLa cells and in viral particles pelleted from the supernatants (Figure 3A). From their apparent molecular weight, major viral proteins in cell lysates were likely the p62 precursor of the envelope glycoprotein E2 (62 kDa), the envelope glycoprotein E1 (52 kDa), and the capsid C (36 kDa) [25]. Other bands in cell lysates probably corresponded to nonstructural and structural proteins, as well as precursors or intermediate cleavage products [2,25,29]. The three main bands in pelleted supernatants likely corresponded to low levels of p62, the E2–E1 doublet (56 and 52 kDa) and C (Figure 3A). A similar Western blot profile was obtained with sera from patients (not shown), indicating that these viral proteins are synthesized and immunogenic in infected individuals.


Characterization of reemerging chikungunya virus.

Sourisseau M, Schilte C, Casartelli N, Trouillet C, Guivel-Benhassine F, Rudnicka D, Sol-Foulon N, Le Roux K, Prevost MC, Fsihi H, Frenkiel MP, Blanchet F, Afonso PV, Ceccaldi PE, Ozden S, Gessain A, Schuffenecker I, Verhasselt B, Zamborlini A, Saïb A, Rey FA, Arenzana-Seisdedos F, Desprès P, Michault A, Albert ML, Schwartz O - PLoS Pathog. (2007)

Assembly and Release of CHIKV(A) Western blot analysis of CHIKV proteins. HeLa cells were exposed to the indicated CHIKV strains (moi 10). Cell lysates and pelleted supernatants were analyzed by Western blot 24 h pi, with a mix of anti-capsid mAb and anti-CHIKV antibodies. The predicted viral proteins are indicated on the right.(B) Gallery of electron micrographs of CHIKV-infected HeLa cells. Cells were analyzed at 36 h pi. CHIKV mostly buds at the plasma membrane of HeLa cells. Viral particles were not detected in noninfected cells (not shown). Bars represent 1 μm in the upper panels and 100 nm in the lower panels.NI, noninfected cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1904475&req=5

ppat-0030089-g003: Assembly and Release of CHIKV(A) Western blot analysis of CHIKV proteins. HeLa cells were exposed to the indicated CHIKV strains (moi 10). Cell lysates and pelleted supernatants were analyzed by Western blot 24 h pi, with a mix of anti-capsid mAb and anti-CHIKV antibodies. The predicted viral proteins are indicated on the right.(B) Gallery of electron micrographs of CHIKV-infected HeLa cells. Cells were analyzed at 36 h pi. CHIKV mostly buds at the plasma membrane of HeLa cells. Viral particles were not detected in noninfected cells (not shown). Bars represent 1 μm in the upper panels and 100 nm in the lower panels.NI, noninfected cells.
Mentions: We also characterized CHIKV by Western blot analysis. Various viral proteins were detected in lysates of CHIKV-infected HeLa cells and in viral particles pelleted from the supernatants (Figure 3A). From their apparent molecular weight, major viral proteins in cell lysates were likely the p62 precursor of the envelope glycoprotein E2 (62 kDa), the envelope glycoprotein E1 (52 kDa), and the capsid C (36 kDa) [25]. Other bands in cell lysates probably corresponded to nonstructural and structural proteins, as well as precursors or intermediate cleavage products [2,25,29]. The three main bands in pelleted supernatants likely corresponded to low levels of p62, the E2–E1 doublet (56 and 52 kDa) and C (Figure 3A). A similar Western blot profile was obtained with sera from patients (not shown), indicating that these viral proteins are synthesized and immunogenic in infected individuals.

Bottom Line: CHIKV replication was cytopathic and associated with an induction of apoptosis in infected cells.CHIKV was highly sensitive to the antiviral activity of type I and II interferons.These results provide a general insight into the interaction between CHIKV and its mammalian host.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Unité Virus et Immunité, Institut Pasteur, Paris, France.

ABSTRACT
An unprecedented epidemic of chikungunya virus (CHIKV) infection recently started in countries of the Indian Ocean area, causing an acute and painful syndrome with strong fever, asthenia, skin rash, polyarthritis, and lethal cases of encephalitis. The basis for chikungunya disease and the tropism of CHIKV remain unknown. Here, we describe the replication characteristics of recent clinical CHIKV strains. Human epithelial and endothelial cells, primary fibroblasts and, to a lesser extent, monocyte-derived macrophages, were susceptible to infection and allowed viral production. In contrast, CHIKV did not replicate in lymphoid and monocytoid cell lines, primary lymphocytes and monocytes, or monocyte-derived dendritic cells. CHIKV replication was cytopathic and associated with an induction of apoptosis in infected cells. Chloroquine, bafilomycin-A1, and short hairpin RNAs against dynamin-2 inhibited viral production, indicating that viral entry occurs through pH-dependent endocytosis. CHIKV was highly sensitive to the antiviral activity of type I and II interferons. These results provide a general insight into the interaction between CHIKV and its mammalian host.

Show MeSH
Related in: MedlinePlus