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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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Cell Tropism of CHIKV(A–C) The indicated human cell lines or primary cells were exposed to CHIKV at an moi of 10. At 24 h pi, cells were fixed, stained with anti-CHIKV antibodies, and analyzed by flow cytometry. The percentage of CHIKV-infected cells is indicated. Data are representative of at least three independent experiments. For primary cells, at least three different donors were analyzed.(A) Examples of sensitive and refractory cell types.(B) Sensitivity of adherent cells to CHIKV infection.(C) Sensitivity of primary blood-derived cells to CHIKV infection. The indicated cell lines, as well as nonactivated PBMCs, activated CD4+ lymphocytes, monocytes, and monocyte-derived DCs were analyzed.
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ppat-0030089-g004: Cell Tropism of CHIKV(A–C) The indicated human cell lines or primary cells were exposed to CHIKV at an moi of 10. At 24 h pi, cells were fixed, stained with anti-CHIKV antibodies, and analyzed by flow cytometry. The percentage of CHIKV-infected cells is indicated. Data are representative of at least three independent experiments. For primary cells, at least three different donors were analyzed.(A) Examples of sensitive and refractory cell types.(B) Sensitivity of adherent cells to CHIKV infection.(C) Sensitivity of primary blood-derived cells to CHIKV infection. The indicated cell lines, as well as nonactivated PBMCs, activated CD4+ lymphocytes, monocytes, and monocyte-derived DCs were analyzed.

Mentions: We tested a panel of immortalized and primary human cells for their ability to replicate CHIKV. The epithelial-derived cell lines HeLa, 293T, and BEAS-2B, as well as primary fibroblasts (Hs 789.Sk skin cells and MRC5 lung cells), were highly susceptible to the virus. After 24 h of infection, more than 60% of the cells were CHIKV-positive by flow cytometry (Figure 4A and 4B) and by immunofluorescence analysis (not shown). CHIKV infection was associated with extensive cell mortality and with the release of high levels of infectious virus in supernatants (not shown). Interestingly, resting MRC5 cells, obtained by maintaining confluent cultures in serum-free medium for at least 10 d before viral exposure (95% of the cells at G0/G1 by flow cytometry analysis of the cell cycle, not shown), were also susceptible to infection (Figure 4B). Therefore, CHIKV replicates in dividing and nondividing cells. We also identified an epithelial cell line (A549 alveolar cells), which was resistant to CHIKV infection. Less than 5% of the cells express CHIKV antigens at day 2 pi, without any CPE (Figure 4A and 4B). We then tested two human endothelial cell lines, TrHBMEC and hCMEC/D3, isolated from the bone marrow and the brain, respectively [32,33]. Interestingly, TrHBMEC cells were readily infected and killed by CHIKV, whereas hCMEC/D3 cells were much more resistant (Figure 4B). With hCMEC/D3, only 1% of the cells were CHIKV+ at 24 h pi, this fraction did not increase over time, and no obvious CPE was observed (not shown). Of note, monkey Vero cells were also sensitive to CHIKV infection (Figure 4B).


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)

Cell Tropism of CHIKV(A–C) The indicated human cell lines or primary cells were exposed to CHIKV at an moi of 10. At 24 h pi, cells were fixed, stained with anti-CHIKV antibodies, and analyzed by flow cytometry. The percentage of CHIKV-infected cells is indicated. Data are representative of at least three independent experiments. For primary cells, at least three different donors were analyzed.(A) Examples of sensitive and refractory cell types.(B) Sensitivity of adherent cells to CHIKV infection.(C) Sensitivity of primary blood-derived cells to CHIKV infection. The indicated cell lines, as well as nonactivated PBMCs, activated CD4+ lymphocytes, monocytes, and monocyte-derived DCs were analyzed.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC1904475&req=5

ppat-0030089-g004: Cell Tropism of CHIKV(A–C) The indicated human cell lines or primary cells were exposed to CHIKV at an moi of 10. At 24 h pi, cells were fixed, stained with anti-CHIKV antibodies, and analyzed by flow cytometry. The percentage of CHIKV-infected cells is indicated. Data are representative of at least three independent experiments. For primary cells, at least three different donors were analyzed.(A) Examples of sensitive and refractory cell types.(B) Sensitivity of adherent cells to CHIKV infection.(C) Sensitivity of primary blood-derived cells to CHIKV infection. The indicated cell lines, as well as nonactivated PBMCs, activated CD4+ lymphocytes, monocytes, and monocyte-derived DCs were analyzed.
Mentions: We tested a panel of immortalized and primary human cells for their ability to replicate CHIKV. The epithelial-derived cell lines HeLa, 293T, and BEAS-2B, as well as primary fibroblasts (Hs 789.Sk skin cells and MRC5 lung cells), were highly susceptible to the virus. After 24 h of infection, more than 60% of the cells were CHIKV-positive by flow cytometry (Figure 4A and 4B) and by immunofluorescence analysis (not shown). CHIKV infection was associated with extensive cell mortality and with the release of high levels of infectious virus in supernatants (not shown). Interestingly, resting MRC5 cells, obtained by maintaining confluent cultures in serum-free medium for at least 10 d before viral exposure (95% of the cells at G0/G1 by flow cytometry analysis of the cell cycle, not shown), were also susceptible to infection (Figure 4B). Therefore, CHIKV replicates in dividing and nondividing cells. We also identified an epithelial cell line (A549 alveolar cells), which was resistant to CHIKV infection. Less than 5% of the cells express CHIKV antigens at day 2 pi, without any CPE (Figure 4A and 4B). We then tested two human endothelial cell lines, TrHBMEC and hCMEC/D3, isolated from the bone marrow and the brain, respectively [32,33]. Interestingly, TrHBMEC cells were readily infected and killed by CHIKV, whereas hCMEC/D3 cells were much more resistant (Figure 4B). With hCMEC/D3, only 1% of the cells were CHIKV+ at 24 h pi, this fraction did not increase over time, and no obvious CPE was observed (not shown). Of note, monkey Vero cells were also sensitive to CHIKV infection (Figure 4B).

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