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A Novel High-Throughput Vaccinia Virus Neutralization Assay and Preexisting Immunity in Populations from Different Geographic Regions in China
- Source :
- PLoS ONE, PLoS ONE, Vol 7, Iss 3, p e33392 (2012)
- Publication Year :
- 2012
- Publisher :
- Public Library of Science (PLoS), 2012.
-
Abstract
- Background Pre-existing immunity to Vaccinia Tian Tan virus (VTT) resulting from a large vaccination campaign against smallpox prior to the early 1980s in China, has been a major issue for application of VTT-vector based vaccines. It is essential to establish a sensitive and high-throughput neutralization assay to understand the epidemiology of Vaccinia-specific immunity in current populations in China. Methodology/Principal Findings A new anti-Vaccinia virus (VACV) neutralization assay that used the attenuated replication-competent VTT carrying the firefly luciferase gene of Photinus pyralis (rTV-Fluc) was established and standardized for critical parameters that included the choice of cell line, viral infection dose, and the infection time. The current study evaluated the maintenance of virus-specific immunity after smallpox vaccination by conducting a non-randomized, cross-sectional analysis of antiviral antibody-mediated immune responses in volunteers examined 30–55 years after vaccination. The rTV-Fluc neutralization assay was able to detect neutralizing antibodies (NAbs) against Vaccinia virus without the ability to differentiate strains of Vaccinia virus. We showed that the neutralizing titers measured by our assay were similar to those obtained by the traditional plaque reduction neutralization test (PRNT). Using this assay, we found a low prevalence of NAb to VTT (7.6%) in individuals born before 1980 from Beijing and Anhui provinces in China, and when present, anti-VTT NAb titers were low. No NAbs were detected in all 222 samples from individuals born after 1980. There was no significant difference observed for titer or prevalence by gender, age range and geographic origin. Conclusion A simplified, sensitive, standardized, reproducible, and high-throughput assay was developed for the quantitation of NAbs against different Vaccinia strains. The current study provides useful insights for the future development of VTT-based vaccination in Beijing and Anhui provinces of China.
- Subjects :
- Male
Viral Diseases
Time Factors
viruses
lcsh:Medicine
Antibodies, Viral
Biochemistry
Neutralization
chemistry.chemical_compound
Mice
Luciferases, Firefly
Smallpox vaccine
lcsh:Science
Mice, Inbred BALB C
Multidisciplinary
Vaccination
Middle Aged
Clinical Laboratory Sciences
Titer
Infectious Diseases
Medicine
Female
Rabbits
Smallpox Vaccine
Research Article
Biotechnology
Adult
China
Adolescent
Immunology
Vaccinia virus
Biology
Microbiology
Virus
Vector Biology
Young Adult
Plaque reduction neutralization test
Immunity
Diagnostic Medicine
Neutralization Tests
Virology
Animals
Humans
lcsh:R
Antibodies, Neutralizing
High-Throughput Screening Assays
Cross-Sectional Studies
chemistry
lcsh:Q
Vaccinia
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 7
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....75b197b7d25c4257816051060b357077
- Full Text :
- https://doi.org/10.1371/journal.pone.0033392