NS1 Protein Secretion during the Acute Phase of West Nile Virus Infection
| Autor: | Brent L. Williams, W. Ian Lipkin, Jessica H. Tonry, Joanne Macdonald, Robert B. Tesh, Mundrigi S. Ashok, Thomas Briese, Omar Jabado, David A. Clark, Roy A. Hall, Gustavo Palacios |
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| Rok vydání: | 2005 |
| Předmět: |
Gene Expression Regulation
Viral medicine.drug_class viruses Immunology Viral Nonstructural Proteins Kidney Monoclonal antibody Microbiology Virus Cell Line Rats Sprague-Dawley Antigen Cricetinae Virology Chlorocebus aethiops medicine Animals Secretion Vero Cells Mesocricetus biology Acute-phase protein virus diseases biochemical phenomena metabolism and nutrition biology.organism_classification Rats Disease Models Animal Kinetics Flavivirus Immunoglobulin M Insect Science biology.protein Pathogenesis and Immunity Antibody West Nile virus West Nile Fever |
| Zdroj: | Journal of Virology. 79:13924-13933 |
| ISSN: | 1098-5514 0022-538X |
| Popis: | The West Nile virus (WNV) nonstructural protein NS1 is a protein of unknown function that is found within, associated with, and secreted from infected cells. We systematically investigated the kinetics of NS1 secretion in vitro and in vivo to determine the potential use of this protein as a diagnostic marker and to analyze NS1 secretion in relation to the infection cycle. A sensitive antigen capture enzyme-linked immunosorbent assay (ELISA) for detection of WNV NS1 (polyclonal-ACE) was developed, as well as a capture ELISA for the specific detection of NS1 multimers (4G4-ACE). The 4G4-ACE detected native NS1 antigens at high sensitivity, whereas the polyclonal-ACE had a higher specificity for recombinant forms of the protein. Applying these assays we found that only a small fraction of intracellular NS1 is secreted and that secretion of NS1 in tissue culture is delayed compared to the release of virus particles. In experimentally infected hamsters, NS1 was detected in the serum between days 3 and 8 postinfection, peaking on day 5, the day prior to the onset of clinical disease; immunoglobulin M (IgM) antibodies were detected at low levels on day 5 postinfection. Although real-time PCR gave the earliest indication of infection (day 1), the diagnostic performance of the 4G4-ACE was comparable to that of real-time PCR during the time period when NS1 was secreted. Moreover, the 4G4-ACE was found to be superior in performance to both the IgM and plaque assays during this time period, suggesting that NS1 is a viable early diagnostic marker of WNV infection. |
| Databáze: | OpenAIRE |
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