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Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis
BACKGROUND: Biting midges of the genus Culicoides vector multiple veterinary pathogens and are difficult to control. Endosymbionts particularly Wolbachia pipientis may offer an alternative to control populations of Culicoides and/or impact disease transmission in the form of population suppression o...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6792224/ https://www.ncbi.nlm.nih.gov/pubmed/31615544 http://dx.doi.org/10.1186/s13071-019-3716-0 |
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author | Ghosh, Arnab Jasperson, Dane Cohnstaedt, Lee W. Brelsfoard, Corey L. |
author_facet | Ghosh, Arnab Jasperson, Dane Cohnstaedt, Lee W. Brelsfoard, Corey L. |
author_sort | Ghosh, Arnab |
collection | PubMed |
description | BACKGROUND: Biting midges of the genus Culicoides vector multiple veterinary pathogens and are difficult to control. Endosymbionts particularly Wolbachia pipientis may offer an alternative to control populations of Culicoides and/or impact disease transmission in the form of population suppression or replacement strategies. METHODS: Culicoides sonorensis cell lines were transfected with a Wolbachia infection using a modified shell vial technique. Infections were confirmed using PCR and cell localization using fluorescent in situ hybridization (FISH). The stability of Wolbachia infections and density was determined by qPCR. qPCR was also used to examine immune genes in the IMD, Toll and JACK/STAT pathways to determine if Wolbachia were associated with an immune response in infected cells. RESULTS: Here we have transfected two Culicoides sonorensis cell lines (W3 and W8) with a Wolbachia infection (walbB) from donor Aedes albopictus Aa23 cells. PCR and FISH showed the presence of Wolbachia infections in both C. sonorensis cell lines. Infection densities were higher in the W8 cell lines when compared to W3. In stably infected cells, genes in the immune Toll, IMD and JAK/STAT pathways were upregulated, along with Attacin and an Attacin-like anti-microbial peptides. CONCLUSIONS: The successful introduction of Wolbachia infections in C. sonorensis cell lines and the upregulation of immune genes, suggest the utility of using Wolbachia for a population replacement and/or population suppression approach to limit the transmission of C. sonorensis vectored diseases. Results support the further investigation of Wolbachia induced pathogen inhibitory effects in Wolbachia-infected C. sonorensis cell lines and the introduction of Wolbachia into C. sonorensis adults via embryonic microinjection to examine for reproductive phenotypes and host fitness effects of a novel Wolbachia infection. |
format | Online Article Text |
id | pubmed-6792224 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-67922242019-10-21 Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis Ghosh, Arnab Jasperson, Dane Cohnstaedt, Lee W. Brelsfoard, Corey L. Parasit Vectors Research BACKGROUND: Biting midges of the genus Culicoides vector multiple veterinary pathogens and are difficult to control. Endosymbionts particularly Wolbachia pipientis may offer an alternative to control populations of Culicoides and/or impact disease transmission in the form of population suppression or replacement strategies. METHODS: Culicoides sonorensis cell lines were transfected with a Wolbachia infection using a modified shell vial technique. Infections were confirmed using PCR and cell localization using fluorescent in situ hybridization (FISH). The stability of Wolbachia infections and density was determined by qPCR. qPCR was also used to examine immune genes in the IMD, Toll and JACK/STAT pathways to determine if Wolbachia were associated with an immune response in infected cells. RESULTS: Here we have transfected two Culicoides sonorensis cell lines (W3 and W8) with a Wolbachia infection (walbB) from donor Aedes albopictus Aa23 cells. PCR and FISH showed the presence of Wolbachia infections in both C. sonorensis cell lines. Infection densities were higher in the W8 cell lines when compared to W3. In stably infected cells, genes in the immune Toll, IMD and JAK/STAT pathways were upregulated, along with Attacin and an Attacin-like anti-microbial peptides. CONCLUSIONS: The successful introduction of Wolbachia infections in C. sonorensis cell lines and the upregulation of immune genes, suggest the utility of using Wolbachia for a population replacement and/or population suppression approach to limit the transmission of C. sonorensis vectored diseases. Results support the further investigation of Wolbachia induced pathogen inhibitory effects in Wolbachia-infected C. sonorensis cell lines and the introduction of Wolbachia into C. sonorensis adults via embryonic microinjection to examine for reproductive phenotypes and host fitness effects of a novel Wolbachia infection. BioMed Central 2019-10-15 /pmc/articles/PMC6792224/ /pubmed/31615544 http://dx.doi.org/10.1186/s13071-019-3716-0 Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Ghosh, Arnab Jasperson, Dane Cohnstaedt, Lee W. Brelsfoard, Corey L. Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
title | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
title_full | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
title_fullStr | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
title_full_unstemmed | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
title_short | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
title_sort | transfection of culicoides sonorensis biting midge cell lines with wolbachia pipientis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6792224/ https://www.ncbi.nlm.nih.gov/pubmed/31615544 http://dx.doi.org/10.1186/s13071-019-3716-0 |
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