Detection of Leishmania RNA Virus in Leishmania Parasites
| Autor: | Nancy G. Saravia, Laurent Philippe Simon Parmentier, Patrick Bastien, Stephen M. Beverley, Francine Pratlong, Florence Prevel, Mary-Anne Hartley, Norbert Müller, Nicolas Fasel, Catherine Ronet, F. Matthew Kuhlmann, Haroun Zangger, John N. Robinson, Patrik Castiglioni, Chantal Desponds |
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| Přispěvatelé: | Université de Lausanne (UNIL), Washington University School of Medicine in St. Louis, Washington University in Saint Louis (WUSTL), Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Centre National de Référence des Leishmanioses [CHRU Montpellier] (CNR-L), Université de Montpellier (UM)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Biologie, Génétique et Pathologie des Pathogènes Eucaryotes (MIVEGEC-BioGEPPE), Pathogènes, Environnement, Santé Humaine (EPATH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Vetsuisse Faculty [Berne, Suisse], University of Bern, Institute Central des Hopitaux Valaisans, Centro Internacional de Entrenamiento e Investigaciones Medicas (CIDEIM), Université de Lausanne = University of Lausanne (UNIL), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Université de Montpellier (UM) |
| Rok vydání: | 2013 |
| Předmět: |
Fluorescent Antibody Technique
Animals Antibodies Monoclonal/diagnostic use Antibodies Viral/diagnostic use Enzyme-Linked Immunosorbent Assay/methods Fluorescent Antibody Technique/methods Humans Immunoblotting/methods Leishmania/virology Mice Mice Inbred C57BL Molecular Sequence Data RNA Viruses/isolation & purification RNA Double-Stranded/immunology RNA Double-Stranded/isolation & purification RNA Viral/genetics Sequence Analysis DNA Virology/methods Dot blot Pathogenesis Antibodies Viral 0302 clinical medicine [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases Leishmania 0303 health sciences 630 Agriculture lcsh:Public aspects of medicine Antibodies Monoclonal 3. Good health Infectious Diseases RNA Viral RNA extraction Antibody Research Article lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 medicine.drug_class Immunoblotting 030231 tropical medicine Enzyme-Linked Immunosorbent Assay Biology Monoclonal antibody Microbiology 03 medical and health sciences Cutaneous leishmaniasis Virology parasitic diseases medicine RNA Viruses [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] RNA Double-Stranded 030304 developmental biology Public Health Environmental and Occupational Health lcsh:RA1-1270 Leishmaniasis RNA virus medicine.disease biology.organism_classification biology.protein Parasitology |
| Zdroj: | PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Public Library of Science, 2013, 7, pp.e2006. ⟨10.1371/journal.pntd.0002006⟩ PLoS neglected tropical diseases PLoS Neglected Tropical Diseases, Vol 7, Iss 1, p e2006 (2013) PLoS Neglected Tropical Diseases, vol. 7, no. 1, pp. e2006 PLoS Neglected Tropical Diseases, 2013, 7, pp.e2006. ⟨10.1371/journal.pntd.0002006⟩ Zangger, Haroun; Ronet, Catherine; Desponds, Chantal; Kuhlmann, F Matthew; Robinson, John; Hartley, Mary-Anne; Prevel, Florence; Castiglioni, Patrik; Pratlong, Francine; Bastien, Patrick; Müller, Norbert; Parmentier, Laurent; Saravia, Nancy Gore; Beverley, Stephen M; Fasel, Nicolas (2013). Detection of Leishmania RNA virus in Leishmania parasites. PLoS neglected tropical diseases, 7(1), e2006. Public Library of Science 10.1371/journal.pntd.0002006 PLOS NEGLECTED TROPICAL DISEASES |
| ISSN: | 1935-2735 1935-2727 |
| DOI: | 10.1371/journal.pntd.0002006 |
| Popis: | Background Patients suffering from cutaneous leishmaniasis (CL) caused by New World Leishmania (Viannia) species are at high risk of developing mucosal (ML) or disseminated cutaneous leishmaniasis (DCL). After the formation of a primary skin lesion at the site of the bite by a Leishmania-infected sand fly, the infection can disseminate to form secondary lesions. This metastatic phenotype causes significant morbidity and is often associated with a hyper-inflammatory immune response leading to the destruction of nasopharyngeal tissues in ML, and appearance of nodules or numerous ulcerated skin lesions in DCL. Recently, we connected this aggressive phenotype to the presence of Leishmania RNA virus (LRV) in strains of L. guyanensis, showing that LRV is responsible for elevated parasitaemia, destructive hyper-inflammation and an overall exacerbation of the disease. Further studies of this relationship and the distribution of LRVs in other Leishmania strains and species would benefit from improved methods of viral detection and quantitation, especially ones not dependent on prior knowledge of the viral sequence as LRVs show significant evolutionary divergence. Methodology/Principal Findings This study reports various techniques, among which, the use of an anti-dsRNA monoclonal antibody (J2) stands out for its specific and quantitative recognition of dsRNA in a sequence-independent fashion. Applications of J2 include immunofluorescence, ELISA and dot blot: techniques complementing an arsenal of other detection tools, such as nucleic acid purification and quantitative real-time-PCR. We evaluate each method as well as demonstrate a successful LRV detection by the J2 antibody in several parasite strains, a freshly isolated patient sample and lesion biopsies of infected mice. Conclusions/Significance We propose that refinements of these methods could be transferred to the field for use as a diagnostic tool in detecting the presence of LRV, and potentially assessing the LRV-related risk of complications in cutaneous leishmaniasis. Author Summary The endosymbiosis of viruses in microbes is a well-described and prevalent environmental partnership, where viruses offer their cellular host incentives of fitness in exchange for the use of their metabolic machinery. We have recently exposed this as an important factor in certain metastatic leishmaniases of South America, where the nucleic acid of a virus residing within some Leishmania parasites acts as a potent innate immunogen causing a destructive inflammatory response, which worsens disease. Leishmania RNA Virus (LRV) exists within many species of Leishmania as a stable infection; these LRV positive strains have been found throughout South America in cutaneous leishmaniases that are often complicated by the occurrence of infectious metastasis with an underlying hyperinflammatory response. In this report, we describe the use of an anti-dsRNA monoclonal antibody (J2), which specifically recognizes dsRNA in a quantitative and sequence-independent fashion. Refined versions of these methods could be transferred to the field as diagnostic tools for detecting the presence of LRV (or other dsRNA viruses), and potentially assessing the LRV-related risk of complicated cutaneous leishmaniasis. |
| Databáze: | OpenAIRE |
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