Mosquito cell-derived West Nile virus replicon particles mimic arbovirus inoculum and have reduced spread in mice
| Autor: | Kristen A. Bernard, Tim W. Carlson, Brendan T. Boylan, Fernando R. Moreira |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
RNA viruses Viral Diseases Physiology viruses Biochemistry Dengue fever Mice Aedes Replicon Pathology and laboratory medicine education.field_of_study Mammalian Genomics lcsh:Public aspects of medicine Hematology Genomics Medical microbiology 3. Good health Enzymes Body Fluids medicine.anatomical_structure Infectious Diseases Blood Arboviral Infections Viruses Female Pathogens Anatomy Oxidoreductases West Nile virus Luciferase Research Article Virus Cultivation lcsh:Arctic medicine. Tropical medicine Arthropoda lcsh:RC955-962 030106 microbiology Population Spleen Biology Arbovirus Infections Transfection Research and Analysis Methods Arbovirus Microbiology Virus Lymphatic System 03 medical and health sciences medicine Genetics Animals Humans education Molecular Biology Techniques Molecular Biology Medicine and health sciences Viral Structural Proteins Biology and life sciences Flaviviruses Host (biology) Inoculation Virus Assembly Public Health Environmental and Occupational Health Organisms Viral pathogens Proteins lcsh:RA1-1270 biochemical phenomena metabolism and nutrition medicine.disease Virology Invertebrates Microbial pathogens Insect Vectors Mice Inbred C57BL 030104 developmental biology Animal Genomics Enzymology Lymph Nodes Arboviruses West Nile Fever |
| Zdroj: | PLOS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Vol 11, Iss 2, p e0005394 (2017) PLoS Neglected Tropical Diseases |
| ISSN: | 1935-2735 |
| DOI: | 10.1371/journal.pntd.0005394 |
| Popis: | Half of the human population is at risk of infection by an arthropod-borne virus. Many of these arboviruses, such as West Nile, dengue, and Zika viruses, infect humans by way of a bite from an infected mosquito. This infectious inoculum is insect cell-derived giving the virus particles distinct qualities not present in secondary infectious virus particles produced by infected vertebrate host cells. The insect cell-derived particles differ in the glycosylation of virus structural proteins and the lipid content of the envelope, as well as their induction of cytokines. Thus, in order to accurately mimic the inoculum delivered by arthropods, arboviruses should be derived from arthropod cells. Previous studies have packaged replicon genome in mammalian cells to produce replicon particles, which undergo only one round of infection, but no studies exist packaging replicon particles in mosquito cells. Here we optimized the packaging of West Nile virus replicon genome in mosquito cells and produced replicon particles at high concentration, allowing us to mimic mosquito cell-derived viral inoculum. These particles were mature with similar genome equivalents-to-infectious units as full-length West Nile virus. We then compared the mosquito cell-derived particles to mammalian cell-derived particles in mice. Both replicon particles infected skin at the inoculation site and the draining lymph node by 3 hours post-inoculation. The mammalian cell-derived replicon particles spread from the site of inoculation to the spleen and contralateral lymph nodes significantly more than the particles derived from mosquito cells. This in vivo difference in spread of West Nile replicons in the inoculum demonstrates the importance of using arthropod cell-derived particles to model early events in arboviral infection and highlights the value of these novel arthropod cell-derived replicon particles for studying the earliest virus-host interactions for arboviruses. Author summary Many emerging viruses of public health concern are arthropod-borne, including tick-borne encephalitis, dengue, Zika, chikungunya, and West Nile viruses. The arboviruses are maintained in nature via virus-specific transmission cycles, involving arthropod (e.g. mosquitos, midges, and ticks) and vertebrate animals (e.g. birds, humans, and livestock). Common to all transmission cycles is the requirement of the arbovirus to replicate in these very different hosts. Since viruses rely on the host cell machinery to produce progeny, the virus particles from these hosts can differ in viral protein glycosylation and lipid content. Thus, the viral inoculum deposited by an infected arthropod will have different properties than virus produced in vertebrate cells. We set out to study the early events of arbovirus infection in a vertebrate host, using the mosquito-borne West Nile virus as a model. Here, we are the first to describe a robust protocol to produce West Nile replicon particles from mosquito cells. Since replicon particles are restricted to a single round of infection, we were able to compare the tropism and spread of the inoculum in animals for mosquito cell- and mammalian cell-derived replicon particles. We found that West Nile replicon particles derived from mosquito cells were significantly reduced in spread to distant sites compared to those derived from mammalian cells. Our results suggest that studies on arbovirus pathogenesis should be conducted with arthropod cell-derived virus, especially for the study of early virus-host interactions. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|