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Native Wolbachia from Aedes albopictus Blocks Chikungunya Virus Infection In Cellulo.

Raquin V, Valiente Moro C, Saucereau Y, Tran FH, Potier P, Mavingui P - PLoS ONE (2015)

Bottom Line: To better understand the Wolbachia/CHIKV/Ae. albopictus interaction, we generated a cellular model using Ae. albopictus derived C6/36 cells that we infected with the wAlbB strain.Our results indicate that CHIKV infection is negatively impacted at both RNA replication and virus assembly/secretion steps in presence of wAlbB.More broadly, this put into question the ecological role of Wolbachia symbiont in Ae. albopictus, but also the ability of the CHIKV to counteract Wolbachia's antiviral potential in vivo.

View Article: PubMed Central - PubMed

Affiliation: Université de Lyon, UMR5557 Ecologie Microbienne, CNRS, USC1190 INRA, VetAgro Sup, Université Lyon 1, Villeurbanne, France.

ABSTRACT
Wolbachia, a widespread endosymbiont of terrestrial arthropods, can protect its host against viral and parasitic infections, a phenotype called "pathogen blocking". However, in some cases Wolbachia may have no effect or even enhance pathogen infection, depending on the host-Wolbachia-pathogen combination. The tiger mosquito Aedes albopictus is naturally infected by two strains of Wolbachia, wAlbA and wAlbB, and is a competent vector for different arboviruses such as dengue virus (DENV) and chikungunya virus (CHIKV). Interestingly, it was shown in some cases that Ae. albopictus native Wolbachia strains are able to inhibit DENV transmission by limiting viral replication in salivary glands, but no such impact was measured on CHIKV replication in vivo. To better understand the Wolbachia/CHIKV/Ae. albopictus interaction, we generated a cellular model using Ae. albopictus derived C6/36 cells that we infected with the wAlbB strain. Our results indicate that CHIKV infection is negatively impacted at both RNA replication and virus assembly/secretion steps in presence of wAlbB. Using FISH, we observed CHIKV and wAlbB in the same mosquito cells, indicating that the virus is still able to enter the cell in the presence of the bacterium. Further work is needed to decipher molecular pathways involved in Wolbachia-CHIKV interaction at the cellular level, but this cellular model can be a useful tool to study the mechanism behind virus blocking phenotype induced by Wolbachia. More broadly, this put into question the ecological role of Wolbachia symbiont in Ae. albopictus, but also the ability of the CHIKV to counteract Wolbachia's antiviral potential in vivo.

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Detection and localization of wAlbB and CHIKV in cellulo by FISH.Detection of Wolbachia 16S rRNA gene (red) and CHIKV Env RNA (green) using oligonucleotide probes labelled with Rhodamine and Alexa488, respectively. Wolbachia signal is detected in C6/36_wAlbB but not in tetracycline treated cells (C6/36_TET). CHIKV signal is detected only in CHIKV infected modality, in the absence or in the presence of Wolbachia where it co-localize with the bacteria in the cytoplasm of C6/36_wAlbB cells. Nuclei of host cells are shown in blue after DAPI labelling (bars = 10 μm).
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pone.0125066.g007: Detection and localization of wAlbB and CHIKV in cellulo by FISH.Detection of Wolbachia 16S rRNA gene (red) and CHIKV Env RNA (green) using oligonucleotide probes labelled with Rhodamine and Alexa488, respectively. Wolbachia signal is detected in C6/36_wAlbB but not in tetracycline treated cells (C6/36_TET). CHIKV signal is detected only in CHIKV infected modality, in the absence or in the presence of Wolbachia where it co-localize with the bacteria in the cytoplasm of C6/36_wAlbB cells. Nuclei of host cells are shown in blue after DAPI labelling (bars = 10 μm).

Mentions: The FISH technique was shown to be an efficient method to detect viruses in mosquito cells [44]. This is the first time such a technique was used to detect CHIKV. The oligonucleotide-probes designed can also detect other alphaviruses, namely Sindbis virus and Ross River virus (not shown). The results showed that CHIKV could be labelled in the cytoplasm of infected cells whereas no CHIKV signal was detected in uninfected cells (Fig 7). Moreover, viral RNA was also detected in cells previously infected with Wolbachia, indicating that at least in some cells the virus is able to penetrate in spite of the presence of the bacterium. However, the co-localization of both Wolbachia and CHIKV was not detected in many cells, and the use of FISH technique did not allowed us to tell if the presence of both micro-organisms in the same host cell was correlated with the load of either bacterium or virus.


Native Wolbachia from Aedes albopictus Blocks Chikungunya Virus Infection In Cellulo.

Raquin V, Valiente Moro C, Saucereau Y, Tran FH, Potier P, Mavingui P - PLoS ONE (2015)

Detection and localization of wAlbB and CHIKV in cellulo by FISH.Detection of Wolbachia 16S rRNA gene (red) and CHIKV Env RNA (green) using oligonucleotide probes labelled with Rhodamine and Alexa488, respectively. Wolbachia signal is detected in C6/36_wAlbB but not in tetracycline treated cells (C6/36_TET). CHIKV signal is detected only in CHIKV infected modality, in the absence or in the presence of Wolbachia where it co-localize with the bacteria in the cytoplasm of C6/36_wAlbB cells. Nuclei of host cells are shown in blue after DAPI labelling (bars = 10 μm).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414612&req=5

pone.0125066.g007: Detection and localization of wAlbB and CHIKV in cellulo by FISH.Detection of Wolbachia 16S rRNA gene (red) and CHIKV Env RNA (green) using oligonucleotide probes labelled with Rhodamine and Alexa488, respectively. Wolbachia signal is detected in C6/36_wAlbB but not in tetracycline treated cells (C6/36_TET). CHIKV signal is detected only in CHIKV infected modality, in the absence or in the presence of Wolbachia where it co-localize with the bacteria in the cytoplasm of C6/36_wAlbB cells. Nuclei of host cells are shown in blue after DAPI labelling (bars = 10 μm).
Mentions: The FISH technique was shown to be an efficient method to detect viruses in mosquito cells [44]. This is the first time such a technique was used to detect CHIKV. The oligonucleotide-probes designed can also detect other alphaviruses, namely Sindbis virus and Ross River virus (not shown). The results showed that CHIKV could be labelled in the cytoplasm of infected cells whereas no CHIKV signal was detected in uninfected cells (Fig 7). Moreover, viral RNA was also detected in cells previously infected with Wolbachia, indicating that at least in some cells the virus is able to penetrate in spite of the presence of the bacterium. However, the co-localization of both Wolbachia and CHIKV was not detected in many cells, and the use of FISH technique did not allowed us to tell if the presence of both micro-organisms in the same host cell was correlated with the load of either bacterium or virus.

Bottom Line: To better understand the Wolbachia/CHIKV/Ae. albopictus interaction, we generated a cellular model using Ae. albopictus derived C6/36 cells that we infected with the wAlbB strain.Our results indicate that CHIKV infection is negatively impacted at both RNA replication and virus assembly/secretion steps in presence of wAlbB.More broadly, this put into question the ecological role of Wolbachia symbiont in Ae. albopictus, but also the ability of the CHIKV to counteract Wolbachia's antiviral potential in vivo.

View Article: PubMed Central - PubMed

Affiliation: Université de Lyon, UMR5557 Ecologie Microbienne, CNRS, USC1190 INRA, VetAgro Sup, Université Lyon 1, Villeurbanne, France.

ABSTRACT
Wolbachia, a widespread endosymbiont of terrestrial arthropods, can protect its host against viral and parasitic infections, a phenotype called "pathogen blocking". However, in some cases Wolbachia may have no effect or even enhance pathogen infection, depending on the host-Wolbachia-pathogen combination. The tiger mosquito Aedes albopictus is naturally infected by two strains of Wolbachia, wAlbA and wAlbB, and is a competent vector for different arboviruses such as dengue virus (DENV) and chikungunya virus (CHIKV). Interestingly, it was shown in some cases that Ae. albopictus native Wolbachia strains are able to inhibit DENV transmission by limiting viral replication in salivary glands, but no such impact was measured on CHIKV replication in vivo. To better understand the Wolbachia/CHIKV/Ae. albopictus interaction, we generated a cellular model using Ae. albopictus derived C6/36 cells that we infected with the wAlbB strain. Our results indicate that CHIKV infection is negatively impacted at both RNA replication and virus assembly/secretion steps in presence of wAlbB. Using FISH, we observed CHIKV and wAlbB in the same mosquito cells, indicating that the virus is still able to enter the cell in the presence of the bacterium. Further work is needed to decipher molecular pathways involved in Wolbachia-CHIKV interaction at the cellular level, but this cellular model can be a useful tool to study the mechanism behind virus blocking phenotype induced by Wolbachia. More broadly, this put into question the ecological role of Wolbachia symbiont in Ae. albopictus, but also the ability of the CHIKV to counteract Wolbachia's antiviral potential in vivo.

Show MeSH
Related in: MedlinePlus