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Zhu R.,National Institutes for Food and Drug Control NIFDC | Zhu R.,Wuhan Institute of Biological Products | Liu Q.,National Institutes for Food and Drug Control NIFDC | Huang W.,National Institutes for Food and Drug Control NIFDC | And 2 more authors.
Archives of Virology | Year: 2014

Vaccinia virus is widely used as a vector in the development of recombinant vaccines. Vaccinia virus strain Guang 9 (VG9), which was derived from vaccinia virus strain Tian Tan (VTT) by successive plaque-cloning purification, was more attenuated than VTT. In this study, the host cell range and the growth and replication of VG9 were compared with those of VTT. The results showed that both VG9 and VTT could infect permissive cells (Vero, TK-143 and CEF) and semipermissive cells PK (15) and induced a visible cytopathic effect (CPE). Both strains could infect nonpermissive CHO-K1 cells but neither was able to reproduce. The replicative ability of VG9 was a little lower than that of VTT. Additionally, recombinant vaccinia viruses containing a firefly luciferase gene (VG9-L and VTT-L) were constructed, and their expression in vitro and replication and spread in vivo were compared. The expression ability of VG9-L was lower than that of VTT-L. Whole-animal imaging data indicated that VG9-L could reproduce quickly and express the exogenous protein at the site of inoculation, regardless of whether the intramuscular, intracutaneous, subcutaneous or celiac inoculation route was used. VG9-L was better in its ability to express a foreign protein than VTT-L, but the time during which expression occurred was shorter. There was no dissemination of virus in mice inoculated with either strain. In summary, this study demonstrates the possibility of using VG9 for the production of smallpox vaccines or the construction of recombinant vaccinia virus vaccines. © 2014, Springer-Verlag Wien.

Xie T.B.,Wuhan Institute of Biological Products
Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi | Year: 2012

To identify and analyze the genetic characteristics of nucleoprotein (N) and glycoprotein (G) genes of rabies virus (RABV) isolated from a donkey in Wuhan. N gene and G gene of the virus were compared with other representative street strains isolated around Hubei areas as well as the vaccine strains used in China and abroad. RABV in brain tissue of a donkey was detected by direct immunofluorescent method and then inoculated in suckling mice to observe the incidence of rabies. Brain samples of the donkey and infected suckling mice were detected by ELISA. The N gene and G gene fragment of the isolated RABV were amplified by RT-PCR and cloned into pMD18-T vector for sequencing and genetic analysis. RABVs were detected in both donkey brain and suckling mice brain samples. The N gene and G gene nucleotide homology of RABV isolated from the donkey with other representative street strains found around Hubei areas as well as vaccine strains used in China and abroad were 85.7% - 99.1% and 82.2% - 99.7%, and the deduced amino acid identity were 95.6% - 99.8% and 87.8% - 99.4%, respectively. Novel RABV was successfully identified and isolated from a donkey and showed close relationship to the representative street strains found around Hubei areas as well as vaccine strains used in China through genetic analysis.

Meng S.-L.,Wuhan Institute of Biological Products
Chinese Journal of Biologicals | Year: 2010

Objective: To analyze the genetic diversity of rabies virus (RV) isolates in China and provide a theoretical basis for prevention of rabies. Methods: The N genes of 26 RV isolates were amplified by RT-PCR and sequenced, and the results were compared with those reported in GenBank, based on which a phylogenetic tree was plotted, and the genotype, genome as well as dynamic evolution in various times and spaces were analyzed. Results: The available Chinese isolates of RV could be divided into two distinct clades, i.e. phylogroup clade I comprising Chinese 1-4 groups and phylogroup clade II comprising Chinese 5-8 groups. The intra-group homologies of nucleotides and amino acids were not less than 93.2% and not less than 94.3%, while the inter-group divergences were not less than 8.0% and not less than 1.7%, respectively. Bayesian Markov chain Monte Carlo (MCMC) analysis suggested that Chinese RV arose in 968 AD, and its annual mean nucleotide substitution rate for N gene was 1. 4089 × 10-4 per site. Conclusion: All the Chinese RV isolates are Lyssavirus genotype 1, which migrate among various regions and reservoir hosts. The isolates of phylogroup clade I are in the same phylogenetic clades with those isolated in the countries in Southeast Asia, such as Thailand, Vietnam, Indonesia, Philippines and Malaysia, while those of phylogroup clade II are found in many parts of the world.

Zhu R.,Wuhan Institute of Biological Products
Chinese Journal of Biologicals | Year: 2011

Objective: To analyze the immunogenicity of attenuated vaccinia virus Guang 9 (VG9) strain. Methods: VG9 and Vaccinia virus Tian Tan (VTT) strains were cultured in chick embryo fibroblasts (CEFs), purified by centrifugation and inoculated i.d. into BALB/c mice. Serum samples were collected once a week for 4 times, and determined for specific antibody titer against vaccinia virus by ELISA. The neutralizing antibody titers in sera 3 and 4 weeks after immunization were determined by neutralization test. Results: Specific antibody titers against vaccinia virus in sera of mice were 1 : 4 050 one week after immunization with both the strains and increased to 1 : 12 150 on week 3. However, the antibody titers against VG9 and VTT strains on week 4 after immunization were 1 : 12 150 and 1 : 36 450 respectively. Both the neutralizing antibody levels induced by the two strains reached peak values 3 weeks after immunization, and cross neutralization was observed. The neutralizing effect on VTT strain was superior to that on VG9 strain, while the ability of VG9 strain in inducing neutralizing antibody was slightly lower than that of VTT strain. Conclusion: Both VG9 and VTT strains showed good immunogenicity, with which the immunization by i. d. route induced specific antibody and neutralizing antibody against vaccinia virus in mice.

Sun Y.,Huazhong University of Science and Technology | Sun Y.,South-Central University for Nationalities | Meng S.,Wuhan Institute of Biological Products
Infection, Genetics and Evolution | Year: 2013

Previous studies showed that DENV-1 transmitted from monkeys to humans approximately 125. years ago. However, there is no comprehensive analysis about phylogeography and population dynamics of Asian DENV-1. Here, we adopt a Bayesian phylogeographic approach to investigate the evolutionary history and phylogeography of Asian DENV-1 using envelope (E) protein gene sequences of 450 viruses isolated from 1954 to 2010 throughout 18 Asian countries and regions. Bayesian phylogeographic analyses indicate that the high rates of viral migration possibly follows long-distance travel for humans in Southeast Asia. Our study highlights that Southeast Asian countries have acted as the main viral sources of the dengue epidemics in East Asia. The results reveal that the time to the most recent common ancestor (TMRCA) of Asian DENV-1 is 1906 (95% HPD, years 1897-1915). We show that the spatial dissemination of virus is the major source of DENV-1 outbreaks in the different localities and leads to subsequent establishment and expansion of the virus in these areas. © 2013 Elsevier B.V.

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