Huo Y.,The Sixth Peoples Hospital of Zhengzhou |
Wan X.,Wuhan Institute of Biological Products |
Ling T.,Wuhan Institute of Biological Products |
Shen S.,Wuhan Institute of Biological Products |
Shen S.,Wuhan Institute of Biological Products Co.
Microbial Pathogenesis | Year: 2016
Noroviruses (NoVs) are the leading cause of non-bacterial acute gastroenteritis worldwide. Due to a lack of cell culture system and animal model, our understanding of NoVs has been lagging behind. In this study, NoV major capsid proteins (VP1) from three different genotypes (GI.2, GII.3 and GII.4) were expressed by using recombinant baculovirus expression system and which led to successful assembly of virus-like particles (VLPs). The receptor binding patterns of three kinds of VLPs were characterized by using synthetic and salivary HBGA-VLP binding assay. Cross-reactivity and cross-blocking activity of rabbit hyperimmune sera against these VLPs were determined by ELISA/Western blot analysis and saliva-VLP binding blockade assay, respectively. Expression of the major capsid proteins from three genotypes all led to smaller VLPs in dominance when sf9 cells were cultured in suspension, which was in consistence with our previous report. These smaller VLPs were used for in vitro synthetic and salivary HBGA-VLP binding and binding blockade assays. VLPs from GII.3 strain exhibited no binding to all synthetic HBGAs and saliva samples tested while VLPs from GI.2 and GII.4 strain showed similar binding pattern and bound to all salivary HBGAs tested. Rabbit anti-GII.3 VLPs hyperimmune serum didn't block the binding of GI.2 and GII.4 VLPs to salivary HBGAs while rabbit anti-GI.2 VLP hyperimmune serum blocked the binding of GII.4 VLPs to salivary HBGAs but not vice versa. Our results provide further evidence indirectly in support of presence of other factors involved in receptor binding other than HBGAs for NoVs, and demonstrate poor cross-blocking activities of antibodies against VLPs within or across genogroups. © 2015 Elsevier Ltd.
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.
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.
Huo Y.,Wuhan Institute of Biological Products |
Wan X.,Wuhan Institute of Biological Products |
Wang Z.,Wuhan Institute of Biological Products |
Meng S.,Wuhan Institute of Biological Products |
Shen S.,Wuhan Institute of Biological Products
Virus Research | Year: 2015
Expression of full-length major capsid protein of Noroviruses (NoVs) in sf9 cells using recombinant baculovirus expression system leads to the formation of virus-like particles (VLPs) with two sizes. In our pursuit of VLPs with uniform sizes, we find that N terminal truncated capsid protein formed primary VLPs with an average size of 21. nm. This kind of VLPs showed similar binding patterns to those produced with full-length major capsid protein. HBGA-VLPs binding assay and saliva-VLPs blocking analysis, as well as stability test demonstrate that the smaller 21. nm VLPs might be an excellent candidate for NoVs vaccine. © 2015 Elsevier B.V.
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.
Gong J.,Huazhong University of Science and Technology |
Liu W.,Huazhong University of Science and Technology |
Zhang J.,Wuhan Institute of Biological Products |
Miao X.,Huazhong University of Science and Technology |
Guo A.-Y.,Huazhong University of Science and Technology
Nucleic Acids Research | Year: 2015
Long non-coding RNAs (lncRNAs) play key roles in various cellular contexts and diseases by diverse mechanisms. With the rapid growth of identified lncRNAs and disease-associated single nucleotide polymorphisms (SNPs), there is a great demand to study SNPs in lncRNAs. Aiming to provide a useful resource about lncRNA SNPs, we systematically identified SNPs in lncRNAs and analyzed their potential impacts on lncRNA structure and function. In total, we identified 495 729 and 777 095 SNPs in more than 30 000 lncRNA transcripts in human and mouse, respectively. A large number of SNPs were predicted with the potential to impact on the miRNA-lncRNA interaction. The experimental evidence and conservation of miRNA-lncRNA interaction, as well as miRNA expressions from TCGA were also integrated to prioritize the miRNA-lncRNA interactions and SNPs on the binding sites. Furthermore, by mapping SNPs to GWAS results, we found that 142 human lncRNA SNPs are GWAS tagSNPs and 197 827 lncRNA SNPs are in the GWAS linkage disequilibrium regions. All these data for human and mouse lncRNAs were imported into IncRNASNP database (http://bioinfo. life.hust.edu.cn/IncRNASNP/), which includes two sub-databases IncRNASNP-human and IncRNASNP-mouse. The IncRNASNP database has a user-friendly interface for searching and browsing through the SNP, lncRNA and miRNA sections. © The Author(s) 2014.
Meng S.L.,Wuhan Institute of Biological Products
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] | Year: 2011
In order to study phylogeography, population dynamics and molecular evolution of rabies viruses (RABVs) isolates from China, especially spatio-temporal dynamics, the timescale of RABVs evolution and its pattern of migration, we performed an extensive comparative analysis of RABV N gene sequence data, representing 167 isolates sampled from 20 provinces in a 78-year period (from 1931 through 2009). The available Chinese isolates could be divided into two distinct clades:Phylogroup clades I comprised Chinese group 1-4; Phylogroup clades II contained Chinese group 5-8. We found no evidence for positive selection (dN/dS>1) acting at any codon and found strong selective constraints for N gene. Bayesian Markov Chain Monte Carlo (MCMC) analysis suggested that the Chinese rabies viruses originated within the last 2000 years and the mean rates of nucleotide substitution for the N gene were approximately 4 x 10(-4) substitutions per site per year. The analyses of the spatial and spatio-temporal evolution indicated that RABV isolates from China migrated among different Provinces.
Chai M.,Wuhan Institute of Biological Products
Zhongguo ji hua mian yi = Chinese journal of vaccines and immunization | Year: 2010
OBJECTIVE: A new Fluorescence Focus Units (FFU) developed for Rabies Virus titration replace current reference method (LD50) so as to reduce the cost. METHODS: The BSR cell was infected by triple serial dilution of the Rabies Virus CTN strain, then detect the fluorescent focus by FITC labeling anti-Rabies Virus monoclonal antibodies. All the samples were assayed in both FFU and LD50. RESULTS: Data generated indicateed that a significant correlation between the two methods (r=99). CONCLUSION: This method is simple and rapid. It will be considered as a useful alternative method to the LD50.