tetano
Editor, Senior Moderator
Vaccine. 2014 Nov 18. pii: S0264-410X(14)01533-3. doi: 10.1016/j.vaccine.2014.11.009. [Epub ahead of print]
The large-scale production of an artificial influenza virus-like particle vaccine in silkworm pupae.
Nerome K1, Sugita S2, Kuroda K3, Hirose T4, Matsuda S4, Majima K5, Kawasaki K6, Shibata T7, Poetri ON8, Soejoedono RD8, Mayasari NL8, Agungpriyono S8, Nerome R4.
Author information
Abstract
We successfully established a mass production system for an influenza virus-like particle (VLP) vaccine using a synthetic H5 hemagglutinin (HA) gene codon-optimized for the silkworm. A recombinant baculovirus containing the synthetic gene was inoculated into silkworm pupae. Four days after inoculation, the hemagglutination titer in homogenates from infected pupae reached a mean value of 0.8 million hemagglutination units (HAU), approximately 2000μg HA protein per pupa, more than 50-fold higher than that produced with an embryonated chicken egg. VLPs ranging from 30nm to 300nm in diameter and covered with a large number of spikes were detected in the homogenates. The spikes were approximately 14nm long, similar to an authentic influenza HA spike. Detailed electron micrographs indicated that the VLP spike density was similar to that of authentic influenza virus particles. The results clearly show that the expression of a single HA gene can efficiently produce VLPs in silkworm pupae. When chickens were immunized with the pupae homogenate, the hemagglutination inhibition titer in their sera reached values of 2048-8192 after approximately 1 month. This is the first report demonstrating that a large amount of VLP vaccine could be produced by single synthetic HA gene in silkworm pupae. Our system might be useful for future vaccine development against other viral diseases.
Copyright ? 2014. Published by Elsevier Ltd.
KEYWORDS:
Baculovirus; H5N1 influenza virus; Vaccine; Virus-like particle
PMID:
25448101
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25448101
The large-scale production of an artificial influenza virus-like particle vaccine in silkworm pupae.
Nerome K1, Sugita S2, Kuroda K3, Hirose T4, Matsuda S4, Majima K5, Kawasaki K6, Shibata T7, Poetri ON8, Soejoedono RD8, Mayasari NL8, Agungpriyono S8, Nerome R4.
Author information
Abstract
We successfully established a mass production system for an influenza virus-like particle (VLP) vaccine using a synthetic H5 hemagglutinin (HA) gene codon-optimized for the silkworm. A recombinant baculovirus containing the synthetic gene was inoculated into silkworm pupae. Four days after inoculation, the hemagglutination titer in homogenates from infected pupae reached a mean value of 0.8 million hemagglutination units (HAU), approximately 2000μg HA protein per pupa, more than 50-fold higher than that produced with an embryonated chicken egg. VLPs ranging from 30nm to 300nm in diameter and covered with a large number of spikes were detected in the homogenates. The spikes were approximately 14nm long, similar to an authentic influenza HA spike. Detailed electron micrographs indicated that the VLP spike density was similar to that of authentic influenza virus particles. The results clearly show that the expression of a single HA gene can efficiently produce VLPs in silkworm pupae. When chickens were immunized with the pupae homogenate, the hemagglutination inhibition titer in their sera reached values of 2048-8192 after approximately 1 month. This is the first report demonstrating that a large amount of VLP vaccine could be produced by single synthetic HA gene in silkworm pupae. Our system might be useful for future vaccine development against other viral diseases.
Copyright ? 2014. Published by Elsevier Ltd.
KEYWORDS:
Baculovirus; H5N1 influenza virus; Vaccine; Virus-like particle
PMID:
25448101
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25448101