Comprehensive mapping of a novel NS1 epitope conserved in flaviviruses within the Japanese encephalitis virus serocomplex
| Autor: | Li-Ping Guo, Rong-Hong Hua, Ye-Nan Li, Hong Huo, Xiao-Lei Wang, Li-Ke Liu, Zhigao Bu, Cheng-Feng Qin |
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| Rok vydání: | 2014 |
| Předmět: |
Cancer Research
medicine.drug_class viruses Molecular Sequence Data Cross Reactions Viral Nonstructural Proteins Dengue virus Antibodies Viral medicine.disease_cause Monoclonal antibody Epitope Virus Flaviviridae Virology medicine Humans Amino Acid Sequence Encephalitis Japanese Conserved Sequence Encephalitis Virus Japanese biology Linear epitope Flavivirus virus diseases biochemical phenomena metabolism and nutrition Japanese encephalitis medicine.disease biology.organism_classification Infectious Diseases Epitope mapping Epitopes B-Lymphocyte Sequence Alignment Epitope Mapping |
| Zdroj: | Virus Research. 185:103-109 |
| ISSN: | 0168-1702 |
| DOI: | 10.1016/j.virusres.2014.03.001 |
| Popis: | Nonstructural protein-1 (NS1) of the Japanese encephalitis virus (JEV) is an immunogenic protein that is a potential candidate for the development of vaccines and diagnostic reagents. NS1 is known to be more specific than the E protein in serological testing of flavivirus infections. However, NS1 exhibits cross-reactivity among flaviviruses even within the same genus and more so within a serocomplex. However, the cross-reactive epitopes on JEV NS1 are poorly characterized. The present study describes the full mapping of a linear B-cell epitope that is common and specific to the JEV serocomplex of Flaviviridae. We generated an NS1-specific monoclonal antibody that cross-reacts with the West Nile virus (WNV) NS1 protein by immunizing mice with recombinant JEV NS1. For epitope mapping, 51 partially overlapping peptides spanning the entire NS1 protein were expressed with a glutathione S-transferase (GST) tag and screened using monoclonal antibodies. Two linear epitope-containing peptides were identified using enzyme-linked immunosorbent assay (ELISA). By sequentially removing amino acid residues from the carboxy and amino terminal of peptides, we successfully identified the smallest unit of the linear epitope required to react with the monoclonal antibody. The linear epitope was located in amino acids residues ²²⁷ETHTLW²³². Furthermore, results of the sequence alignment revealed that the epitope was highly conserved among JEV strains. Notably, the epitope is highly conserved among viruses of the JEV serocomplex. Furthermore, the homologous regions on NS1 proteins from dengue viruses showed no cross-reactivity with the monoclonal antibodies. The epitope was recognized by antisera against the WNV but not against the dengue virus. This novel JEV serocomplex-specific linear B-cell epitope of NS1 would be helpful in the development of new vaccines and diagnostic assays. |
| Databáze: | OpenAIRE |
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