National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control

Guangzhou, China

National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control

Guangzhou, China
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FENG M.,South China Agricultural University | FENG M.,National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control | FENG M.,Key Laboratory of Veterinary Vaccine Innovation | FENG M.,Key Laboratory of Zoonosis Prevention and Control of Guangdong Province | And 12 more authors.
Journal of Integrative Agriculture | Year: 2017

Rapid diagnostic methods for classifying avian leukosis subgroups in the field were needed for routine, large-scale screening. As a first step in method development, we inserted the avian leukosis virus subgroup A (ALV-A) env gene into plasmid pcDNA3.1/Zeo (+) and used this construct to transfect DF-1 cells. Zeocin-resistant cells were obtained after 2 weeks of zeocin selection. Then, the cells were analyzed using PCR, immunofluorescence, and Western blot for expression of the envA-encoded envelope protein after 30 serial passages. The DF-1/A cell line was completely resistant to 104 TCID50/0.1 mL (50% tissue culture infective dose) ALV-A and was partially resistant to 105 TCID50/0.1 mL ALV-A viral particles. By comparing the DF-1/A and DF-1 cell lines, an ALV-A isolate was identified using a gag-specific ELISA for capsid protein p27. Thus, we established a DF-1/A cell line that was resistant to ALV-A infection. This cell line will be useful as a diagnostic tool. © 2017 CAAS. Publishing services by Elsevier B.V


Wei L.,National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control | Wei L.,Key Laboratory of Animal Vaccine Development | Wei L.,Key Laboratory of Zoonosis Prevention and Control of Guangdong | Wei L.,South China Agricultural University | And 34 more authors.
Veterinary Immunology and Immunopathology | Year: 2013

In mammals, Toll-like receptor 7 (TLR7) is an important membrane-bound receptor triggered by antiviral compounds and single-stranded RNA. It is implicated in the immune response to viruses such as influenza virus. It was not known whether geese, a natural host for avian influenza viruses, possess a homologue of mammalian TLR7 for recognizing avian influenza virus. In this study, we cloned the full-length of goose TLR7 and partial sequences of its adaptor protein, myeloid differentiation factor 88 (MyD88), some antiviral molecules such as RNA-dependent protein kinase (PKR) and 2',5'-oligoadenylate synthetase (OAS). Goose TLR7 has a protein secondary structure identical to that of mammals, consisting of several leucine-rich domains, a transmembrane domain, and Toll/interleukin-1 receptor domain. To further understand whether the MyD88-dependent pathway of TLR7 is involved in the antiviral innate immune response against highly pathogenic avian influenza virus (HPAIV) infection in geese, we inoculated geese with an H5N1 HPAIV isolated from ducks in 2004. The virus, A/Duck/Guangdong/212/2004, replicated in various tissues resulting in 40% mortality. Quantitative real-time PCR analysis showed upregulation of mRNA transcripts for TLR7, MyD88, PKR and OAS in the lungs of geese at 1, 2 and 3 days post-inoculation. Therefore, the MyD88-dependent pathway of TLR7 was involved in the early stage of antiviral innate immune response in geese during H5N1 HPAIV infection. © 2013.


Cui J.,South China Agricultural University | Cui J.,National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control | Cui J.,CAS Shenyang Institute of Metal Research | Qu N.,South China Agricultural University | And 23 more authors.
Frontiers in Cellular and Infection Microbiology | Year: 2017

We analyzed five H5N1 avian influenza viruses (AIVs) isolated from different birds in 2012 in China. Based on whole-genome sequences, we divided the viruses into four genotypes. The DKE26, GSE43, and DKE53 viruses belonged to Genotypes 1–3, respectively. The CKE93 and CKE96 viruses were classified into Genotype 4. Genotypes 1–3 correspond to the viruses containing the HA gene of clade 2.3.2, and Genotype 4 is the virus that bears the HA gene of clade 7.2. To better understand the pathogenicity and transmission of the viruses, we infected chickens with 103 EID50 /0.1 ml GSE43 (clade 2.3.2) or CKE93 (clade 7.2) virus. Our results revealed that 6 of 7 specific-pathogen-free (SPF) chickens inoculated with GSE43 virus were dead before 7-day post-infection, but all the SPF chickens inoculated with CKE93 virus survived the infection. Both the GSE43 and CKE93 viruses replicated systemically in chickens. The virus titers of GSE43 virus in tested organs were obviously higher than those of CKE93 virus. Our results revealed that the pathogenicity and replication of GSE43 in chickens was much higher than those of CKE93. The GSE43 virus could transmit between chickens, but the CKE93 could not transmit between chickens by naïve contact. Therefore, different clades of H5N1 AIVs possessed variable pathogenicities and transmission abilities among chickens. Our study contributes to knowledge of pathogenic variations of prevalent H5N1 viruses. © 2017 Cui, Qu, Guo, Cao, Wu, Mei, Sun, Lu, Qin, Jiao and Liao.


Zhang Z.,South China Agricultural University | Zhang Z.,National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control | Zhang Z.,Key Laboratory of Animal Vaccine Development | Zhang Z.,Key Laboratory of Zoonosis Prevention and Control of Guangdong Province | And 26 more authors.
Veterinary Microbiology | Year: 2017

H5N1, a highly pathogenic avian influenza virus (HPAIV), poses a significant threat to poultry and human health. However, currently available inactivated influenza vaccines are less efficacious against viruses that display antigenic drift. In this study, we constructed a recombinant baculovirus (BV-HMNN) expressing four conserved antigen epitopes: H5N1 hemagglutinin stem area amino acids 76–130 (HA2 76–130); three tandem repeats from the ectodomain of the conserved influenza matrix protein M2 (3M2e); nucleoprotein amino acids 55–69 (NP55–69); and nucleoprotein amino acids 380–393 (NP380–393). We evaluated the immunogenicity and protective efficacy of coimmunization with an inactivated avian influenza virus vaccine (Re6) and the recombinant baculovirus (BV-HMNN) against heterologous viral infection in specific-pathogen-free chickens. The chickens immunized with both vaccines (Re6 + BV-HMNN) achieved complete protection, was significantly greater than that of chickens vaccinated with Re6 alone. BV-HMNN-supplemented vaccination also reduced viral shedding more effectively than nonsupplemented vaccination. We conclude that coimmunization with both vaccines was superior to immunization with the inactivated vaccine alone in inducing cross-protection against heterologous H5N1 virus. © 2017

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