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ABSTRACT Multicellular organisms evolved sophisticated defence systems to confer protection against pathogens. An important characteristic of these immune systems is their ability to act
both locally at the site of infection and at distal uninfected locations1,2,3,4. In insects, such as _Drosophila melanogaster,_ RNA interference (RNAi) mediates antiviral immunity5,6,7.
However, the antiviral RNAi defence in flies seems to be a local, cell-autonomous process, as flies are thought to be unable to generate a systemic RNAi response8. Here we show that a
recently defined double-stranded RNA (dsRNA) uptake pathway9 is essential for effective antiviral RNAi immunity in adult flies. Mutant flies defective in this dsRNA uptake pathway were
hypersensitive to infection with _Drosophila_ C virus and Sindbis virus. Mortality in dsRNA-uptake-defective flies was accompanied by 100-to 105-fold increases in viral titres and higher
levels of viral RNA. Furthermore, inoculating naked dsRNA into flies elicited a sequence-specific antiviral immune response that required an intact dsRNA uptake pathway. These findings
suggest that spread of dsRNA to uninfected sites is essential for effective antiviral immunity. Notably, infection with green fluorescent protein (GFP)-tagged Sindbis virus suppressed
expression of host-encoded GFP at a distal site. Thus, similar to protein-based immunity in vertebrates, the antiviral RNAi response in flies also relies on the systemic spread of a
virus-specific immunity signal. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS Access through your
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FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS INNATE IMMUNE PATHWAYS ACT SYNERGISTICALLY TO CONSTRAIN RNA VIRUS EVOLUTION IN _DROSOPHILA MELANOGASTER_ Article 10
March 2022 RETROTRANSPOSON ACTIVATION DURING _DROSOPHILA_ METAMORPHOSIS CONDITIONS ADULT ANTIVIRAL RESPONSES Article 17 November 2022 TWO CGAS-LIKE RECEPTORS INDUCE ANTIVIRAL IMMUNITY IN
_DROSOPHILA_ Article 14 July 2021 REFERENCES * Baulcombe, D. RNA silencing in plants. _Nature_ 431, 356–363 (2004) Article ADS CAS Google Scholar * Dorner, T. & Radbruch, A.
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CAS Google Scholar Download references ACKNOWLEDGEMENTS We are grateful to members of the Andino and the O’Farrell laboratories for discussion, technical support and advice on fly work.
We thank J. Frydman, P. O’Farrell and M. Vignuzzi for discussions and comments on the manuscript. We also thank T. Cook for advice on the IR _EcR_ eye phenotype, R. Carthew for providing the
GMR>IR[_white_] and _Dcr2_ fly stocks and M. Siomi for the _Ago2_ fly stock. B.G. is a Manlio Cantarini fellow. B.B. is a Lebanese CNRSL Fellow. C.J. is a University of Paris VI and
Ministère de la Recherche fellow. This work was financially supported by NIH grants AI40085 and AI064738 to R.A., the Institut Pasteur to M.-C.S. and C.A., and CNRS, ANR and ARC grants to
C.A. AUTHOR CONTRIBUTIONS M.-C.S., M.T. and R.P.v.R. performed dsRNA inoculations and virus infections, normal and reverse northern blotting, western blotting, survival curves, obtained
fluorescent images, and prepared and analysed mutant flies. B.G. and V.G. examined systemic spread of dsRNA. The genetic and phenotypic analyses of transgenic flies expressing RNA hairpins
were designed and carried out by B.B., C.J. and C.A. M.-C.S. and R.A. designed the experiments, discussed the interpretation of the results and co-wrote the manuscript. AUTHOR INFORMATION
Author notes * Maria-Carla Saleh & Ronald P. van Rij Present address: Present addresses: Institut Pasteur, Viruses and RNA interference, F-75015 Paris, France (M.-C.S.); Department of
Medical Microbiology, Nijmegen Center for Molecular Life Sciences, University Medical Center Nijmegen, PO Box 9101, 6500 HB Nijmegen, The Netherlands (R.P.v.R.)., * Michel Tassetto and
Ronald P. van Rij: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Department of Microbiology and Immunology, University of California, San Francisco 94122-2280,
USA, Maria-Carla Saleh, Michel Tassetto, Ronald P. van Rij & Raul Andino * Institut Pasteur, Viruses and RNA interference, F-75015 Paris, France, Bertsy Goic & Valérie Gausson *
Institut Pasteur, Drosophila Genetics and Epigenetics; CNRS, URA 2578, F-75015 Paris, France, Bassam Berry, Caroline Jacquier & Christophe Antoniewski * CNRS, URA 2578, F-75015 Paris,
France, Bassam Berry, Caroline Jacquier & Christophe Antoniewski Authors * Maria-Carla Saleh View author publications You can also search for this author inPubMed Google Scholar * Michel
Tassetto View author publications You can also search for this author inPubMed Google Scholar * Ronald P. van Rij View author publications You can also search for this author inPubMed
Google Scholar * Bertsy Goic View author publications You can also search for this author inPubMed Google Scholar * Valérie Gausson View author publications You can also search for this
author inPubMed Google Scholar * Bassam Berry View author publications You can also search for this author inPubMed Google Scholar * Caroline Jacquier View author publications You can also
search for this author inPubMed Google Scholar * Christophe Antoniewski View author publications You can also search for this author inPubMed Google Scholar * Raul Andino View author
publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Raul Andino. SUPPLEMENTARY INFORMATION SUPPLEMENTARY FIGURES This file
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FIG. 4 POWERPOINT SLIDE FOR FIG. 5 RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Saleh, MC., Tassetto, M., van Rij, R. _et al._ Antiviral immunity in
_Drosophila_ requires systemic RNA interference spread. _Nature_ 458, 346–350 (2009). https://doi.org/10.1038/nature07712 Download citation * Received: 04 October 2008 * Accepted: 03
December 2008 * Published: 08 February 2009 * Issue Date: 19 March 2009 * DOI: https://doi.org/10.1038/nature07712 SHARE THIS ARTICLE Anyone you share the following link with will be able to
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