Chapter title |
Generation of a Single-Cycle Replicable Rift Valley Fever Vaccine.
|
---|---|
Chapter number | 9 |
Book title |
Vaccine Design
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3387-7_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3385-3, 978-1-4939-3387-7
|
Authors |
Shin Murakami, Kaori Terasaki, Shinji Makino |
Editors |
Sunil Thomas |
Abstract |
Rift Valley fever virus (RVFV) (genus Phlebovirus, family Bunyaviridae) is an arbovirus that causes severe disease in humans and livestock in sub-Saharan African countries. The virus carries a tripartite, single-stranded, and negative-sense RNA genome, designated as L, M, and S RNAs. RVFV spread can be prevented by the effective vaccination of animals and humans. Although the MP-12 strain of RVFV is a live attenuated vaccine candidate, MP-12 showed neuroinvasiveness and neurovirulence in young mice and immunodeficiency mice. Hence, there is a concern for the use of MP-12 to certain individuals, especially those that are immunocompromised. To improve MP-12 safety, we have generated a single-cycle, replicable MP-12 (scMP-12), which carries L RNA, S RNA encoding green fluorescent protein in place of a viral nonstructural protein NSs, and an M RNA encoding a mutant envelope protein lacking an endoplasmic reticulum retrieval signal and defective for membrane fusion function. The scMP-12 undergoes efficient amplification in the Vero-G cell line, which is a Vero cell line stably expressing viral envelope proteins, while it undergoes single-cycle replication in naïve cells and completely lacks neurovirulence in suckling mice after intracranial inoculation. A single-dose vaccination of mice with scMP-12 confers protective immunity. Thus, scMP-12 represents a new, promising RVF vaccine candidate. Here we describe protocols for scMP-12 generation by using a reverse genetics system, establishment of Vero-G cells, and titration of scMP-12 in Vero-G cells. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 10 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 1 | 10% |
Professor > Associate Professor | 1 | 10% |
Researcher | 1 | 10% |
Other | 1 | 10% |
Unknown | 6 | 60% |
Readers by discipline | Count | As % |
---|---|---|
Immunology and Microbiology | 2 | 20% |
Veterinary Science and Veterinary Medicine | 1 | 10% |
Agricultural and Biological Sciences | 1 | 10% |
Biochemistry, Genetics and Molecular Biology | 1 | 10% |
Unknown | 5 | 50% |