Recombinant dengue 2 virus NS3 protein conserves structural antigenic and immunological properties relevant for dengue vaccine design
| العنوان: | Recombinant dengue 2 virus NS3 protein conserves structural antigenic and immunological properties relevant for dengue vaccine design |
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| المؤلفون: | Emiliana M. Silva, María G. Guzmán, Alienys Izquierdo, Mayra Muné, Ronaldo Mohana-Borges, Ana B. Pérez, Mayling Alvarez, Oney Ortega, Yudira Soto, Angélica García, Rosa Ramírez, Rosabel Falcón |
| المصدر: | Virus Genes. 49:185-195 |
| بيانات النشر: | Springer Science and Business Media LLC, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | Antigenicity, viruses, Blotting, Western, Fluorescent Antibody Technique, Gene Expression, Dengue Vaccines, Enzyme-Linked Immunosorbent Assay, Viral Nonstructural Proteins, Dengue virus, Antibodies, Viral, medicine.disease_cause, Epitope, Interferon-gamma, Virology, Protein A/G, Escherichia coli, Genetics, medicine, Animals, Cloning, Molecular, Molecular Biology, Dengue vaccine, Mice, Inbred BALB C, Vaccines, Synthetic, NS3, biology, Tumor Necrosis Factor-alpha, Serine Endopeptidases, virus diseases, General Medicine, Dengue Virus, biochemical phenomena, metabolism, and nutrition, Fusion protein, Recombinant Proteins, Interleukin-10, Leukocytes, Mononuclear, biology.protein, Female, RNA Helicases, Myc-tag |
| الوصف: | The NS3 protein is a multifunctional non-structural protein of flaviviruses implicated in the polyprotein processing. The predominance of cytotoxic T cell lymphocytes epitopes on the NS3 protein suggests a protective role of this protein in limiting virus replication. In this work, we studied the antigenicity and immunogenicity of a recombinant NS3 protein of the Dengue virus 2. The full-length NS3 gene was cloned and expressed as a His-tagged fusion protein in Escherichia coli. The pNS3 protein was purified by two chromatography steps. The recombinant NS3 protein was recognized by anti-protease NS3 polyclonal antibody and anti-DENV2 HMAF by Western Blot. This purified protein was able to stimulate the secretion of high levels of gamma interferon and low levels of interleukin-10 and tumor necrosis factor-α in mice splenocytes, suggesting a predominantly Th-1-type T cell response. Immunized BALB/c mice with the purified NS3 protein showed a strong induction of anti-NS3 IgG antibodies, essentially IgG2b, as determined by ELISA. Immunized mice sera with recombinant NS3 protein showed specific recognition of native dengue protein by Western blotting and immunofluorescence techniques. The successfully purified recombinant protein was able to preserv the structural and antigenic determinants of the native dengue protein. The antigenicity shown by the recombinant NS3 protein suggests its possible inclusion into future DENV vaccine preparations. |
| تدمد: | 1572-994X 0920-8569 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a0edf82152256dfc9e0f7edc80c1a1a2Test https://doi.org/10.1007/s11262-014-1087-3Test |
| حقوق: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....a0edf82152256dfc9e0f7edc80c1a1a2 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1572994X 09208569 |
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