West Nile virus recombinant DNA vaccine protects mouse and horse from virus challenge and expresses in vitro a noninfectious recombinant antigen that can be used in enzyme-linked immunosorbent assays.
Abstract: Introduction of West Nile (WN) virus into the United States in 1999 created major human and animal health concerns. Currently, no human or veterinary vaccine is available to prevent WN viral infection, and mosquito control is the only practical strategy to combat the spread of disease. Starting with a previously designed eukaryotic expression vector, we constructed a recombinant plasmid (pCBWN) that expressed the WN virus prM and E proteins. A single intramuscular injection of pCBWN DNA induced protective immunity, preventing WN virus infection in mice and horses. Recombinant plasmid-transformed COS-1 cells expressed and secreted high levels of WN virus prM and E proteins into the culture medium. The medium was treated with polyethylene glycol to concentrate proteins. The resultant, containing high-titered recombinant WN virus antigen, proved to be an excellent alternative to the more traditional suckling-mouse brain WN virus antigen used in the immunoglobulin M (IgM) antibody-capture and indirect IgG enzyme-linked immunosorbent assays. This recombinant antigen has great potential to become the antigen of choice and will facilitate the standardization of reagents and implementation of WN virus surveillance in the United States and elsewhere.
Publication Date: 2001-04-05 PubMed ID: 11287553PubMed Central: PMC114149DOI: 10.1128/JVI.75.9.4040-4047.2001Google Scholar: Lookup
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Summary
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The research conducted aimed to create a vaccine capable of preventing West Nile (WN) viral infection in animals such as mice and horses using a recombinant DNA technique. In addition, the researchers also developed a noninfectious recombinant antigen for use in diagnostic tests.
Objective and Background of the Research
- West Nile virus was introduced into the United States in 1999 and caused significant health concerns for both humans and animals. At the time of this research, there were no vaccines available to prevent infection. The control of mosquitoes, who transit the disease, was the main strategy implemented to try and prevent spread of this virus.
- The researchers decided to attempt to develop a vaccine using a previously designed genetic expression system. This involved constructing a recombinant plasmid (a small circular piece of DNA which can be replicated independently of the host organism) that expressed key proteins from the WN virus that are capable of inducing immunity against the virus.
Creation of the Vaccine
- A recombinant plasmid, pCBWN, was developed, which was able to produce two key proteins from the WN virus: the precursor of the membrane protein (prM) and the envelope glycoprotein (E).
- An intramuscular injection of pCBWN was given to mice and horses. This technique is designed to stimulate the recipient’s own immune system to recognise these proteins as foreign and mount an immune response against them, thus providing immunity against the actual virus should the individual become infected.
- The outcome demonstrated that the vaccine was successful in providing immunity, thereby preventing WN virus infection in the animals tested.
Development of the Recombinant Antigen
- In addition to the vaccine, the researchers created a noninfectious recombinant antigen (proteins that can stimulate an immune response) in vitro (in a lab environment).
- This was achieved by transforming COS-1 cells (a cell line derived from monkey kidney tissue) with the recombinant plasmid. These cells were able to produce and secrete high levels of the WN virus prM and E proteins.
- This noninfectious recombinant antigen proved to be an excellent alternative for use in diagnostic tests – specifically enzyme-linked immunosorbent assays (ELISA). These assays are used to detect and measure antibodies in blood which can confirm exposure or immunity to a certain pathogen.
- Traditionally, the antigen used in these tests was derived from the brains of suckling mice. The use of the recombinant antigen offers a more ethical and potentially safer alternative. The standardization of this antigen could potentially improve the enactment of WN virus surveillance in the United States and globally.
Cite This Article
APA
Davis BS, Chang GJ, Cropp B, Roehrig JT, Martin DA, Mitchell CJ, Bowen R, Bunning ML.
(2001).
West Nile virus recombinant DNA vaccine protects mouse and horse from virus challenge and expresses in vitro a noninfectious recombinant antigen that can be used in enzyme-linked immunosorbent assays.
J Virol, 75(9), 4040-4047.
https://doi.org/10.1128/JVI.75.9.4040-4047.2001 Publication
Researcher Affiliations
- Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Fort Collins, Colorado 80522, USA.
MeSH Terms
- Amino Acid Sequence
- Animals
- Antigens, Viral / biosynthesis
- Antigens, Viral / genetics
- Antigens, Viral / immunology
- COS Cells
- Chlorocebus aethiops
- Enzyme-Linked Immunosorbent Assay / methods
- Female
- Gene Expression
- Horses
- Mice
- Mice, Inbred ICR
- Molecular Sequence Data
- Plasmids
- Vaccines, DNA / genetics
- Vaccines, DNA / immunology
- Viral Envelope Proteins / biosynthesis
- Viral Envelope Proteins / genetics
- Viral Envelope Proteins / immunology
- Viral Vaccines / genetics
- Viral Vaccines / immunology
- West Nile Fever / diagnosis
- West Nile Fever / immunology
- West Nile Fever / prevention & control
- West Nile Fever / virology
- West Nile virus / genetics
- West Nile virus / immunology
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