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ABSTRACT: Nipah virus (NiV), a member of the Paramyxoviridae family, causes deadly encephalitis in humans and huge economic losses to the pig industry. Here, we generated recombinant avirulent Newcastle disease virus (NDV) LaSota strains expressing the NiV G and F proteins respectively (designated as rLa-NiVG and rLa-NiVF), and evaluated their immunogenicity in mice and pigs. Both rLa-NiVG and rLa-NiVF displayed growth properties similar to those of LaSota virus in chicken eggs. Co-infection of rLa-NiVG and rLa-NiVF caused marked syncytia formation, while intracerebral co-inoculation of these viruses in mice showed they were safe in at least one mammalian species. Animal immunization studies showed rLa-NiVG and rLa-NiVF induced NiV neutralizing antibody responses in mice and pigs, and F protein-specific CD8+ T cell responses in mice. Most importantly, rLa-NiVG and rLa-NiVF administered alone or together, induced a long-lasting neutralizing antibody response in pigs. Recombinant rLa-NiVG/F thus appear to be promising NiV vaccine candidates for pigs and potentially humans.
Virology 06/2012; 432(2):327-35. · 3.35 Impact Factor
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ABSTRACT: Effective, safe, and affordable rabies vaccines are still being sought. Attenuated live vaccine has been widely used to protect carnivores from canine distemper. In this study, we generated a recombinant canine distemper virus (CDV) vaccine strain, rCDV-RVG, expressing the rabies virus glycoprotein (RVG) by using reverse genetics. The recombinant virus rCDV-RVG retained growth properties similar to those of vector CDV in Vero cell culture. Animal studies demonstrated that rCDV-RVG was safe in mice and dogs. Mice inoculated intracerebrally or intramuscularly with rCDV-RVG showed no apparent signs of disease and developed a strong rabies virus (RABV) neutralizing antibody response, which completely protected mice from challenge with a lethal dose of street virus. Canine studies showed that vaccination with rCDV-RVG induced strong and long-lasting virus neutralizing antibody responses to RABV and CDV. This is the first study demonstrating that recombinant CDV has the potential to serve as bivalent live vaccine against rabies and canine distemper in animals.
Vaccine 06/2012; 30(34):5067-72. · 3.77 Impact Factor
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ABSTRACT: The rabies Flury Low Egg Passage virus (LEP) has been widely used as a seed virus to generate inactive vaccine. Here, we established a reverse genetic system for LEP and generated a recombinant LEP virus (rLEP-G) that carries two identical G genes. This recombinant virus showed similar properties to those of LEP with respect to in vitro growth, neurotropism index, and virulence in mice. rLEP-G produced 4.3-fold more G protein than did LEP in BHK-21 cells. The inactivated vaccine generated from rLEP-G induced significantly higher virus neutralization titers in mice and dogs than those produced in response to LEP-derived vaccine. Our results suggest that rLEP-G is an improved seed virus candidate for inactivated rabies virus vaccine manufacture.
Virology Journal 09/2011; 8:454. · 2.34 Impact Factor
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ABSTRACT: Ducks play an important role in the maintenance of highly pathogenic H5N1 avian influenza viruses (AIVs) in nature, and the successful control of AIVs in ducks has important implications for the eradication of the disease in poultry and its prevention in humans. The inactivated influenza vaccine is expensive, labor-intensive, and usually needs 2 to 3 weeks to induce protective immunity in ducks. Live attenuated duck enteritis virus (DEV; a herpesvirus) vaccine is used routinely to control lethal DEV infections in many duck-producing areas. Here, we first established a system to generate the DEV vaccine strain by using the transfection of overlapping fosmid DNAs. Using this system, we constructed two recombinant viruses, rDEV-ul41HA and rDEV-us78HA, in which the hemagglutinin (HA) gene of the H5N1 virus A/duck/Anhui/1/06 was inserted and stably maintained within the ul41 gene or between the us7 and us8 genes of the DEV genome. Duck studies indicated that rDEV-us78HA had protective efficacy similar to that of the live DEV vaccine against lethal DEV challenge; importantly, a single dose of 10(6) PFU of rDEV-us78HA induced complete protection against a lethal H5N1 virus challenge in as little as 3 days postvaccination. The protective efficacy against both lethal DEV and H5N1 challenge provided by rDEV-ul41HA inoculation in ducks was slightly weaker than that provided by rDEV-us78HA. These results demonstrate, for the first time, that recombinant DEV is suitable for use as a bivalent live attenuated vaccine, providing rapid protection against both DEV and H5N1 virus infection in ducks.
Journal of Virology 08/2011; 85(21):10989-98. · 5.40 Impact Factor
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ABSTRACT: To study the biological characteristics and pathogenicity of a recombinant rabies virus Flury LEP (low egg passage) that has two glycoprotein genes (G gene).
By using reverse genetics techniques, we constructed a recombinant virus Flury LEP that has an additional G gene between P and M gene (rLEP-PGM). Then we studied the biological characteristics of the recombinant virus and its pathogenicity on mice.
The in vitro growth characteristic of rLEP-PGM were similar to the LEP strain. Western blot analysis of glycoprotein expression showed that the glycoprotein expression level of rLEP-PGM was 1.5 times higher than LEP. The LD50 of rLEP-PGM and LEP was 3 FFU and 1 FFU by intracerebral injection. However, the LD50 of intramuscular injection was 4 x 10(4) Lg FFU and 3.2 x 10(5) Lg FFU, respectively.
Insertion of an additional G gene between P and M gene can significantly raise the expression level of glycoprotein and enhance the ability to invade central nervous system from peripheral sites.
ACTA MICROBIOLOGICA SINICA 08/2011; 51(8):1098-105.
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ABSTRACT: Effective, safe, and affordable rabies vaccines are still being sought. Newcastle disease virus (NDV), an avian paramyxovirus, has shown promise as a vaccine vector for mammals. Here, we generated a recombinant avirulent NDV La Sota strain expressing the rabies virus glycoprotein (RVG) and evaluated its potential to serve as a vaccine against rabies. The recombinant virus, rL-RVG, retained its high-growth property in chicken eggs, with titers of up to 10⁹·⁸ 50% egg infective doses (EID₅₀)/ml of allantoic fluid. RVG expression enabled rL-RVG to spread from cell to cell in a rabies virus-like manner, and RVG was incorporated on the surface of the rL-RVG viral particle. RVG incorporation did not alter the trypsin-dependent infectivity of the NDV vector in mammalian cells. rL-RVG and La Sota NDV showed similar levels of sensitivity to a neutralization antibody against NDV and similar levels of resistance to a neutralization antibody against rabies virus. Animal studies demonstrated that rL-RVG is safe in several species, including cats and dogs, when administered as multiple high doses of recombinant vaccine. Intramuscular vaccination with rL-RVG induced a substantial rabies virus neutralization antibody response and provided complete protection from challenge with circulating rabies virus strains. Most importantly, rL-RVG induced strong and long-lasting protective neutralization antibody responses to rabies virus in dogs and cats. A low vaccine dose of 10⁸·³ EID₅₀ completely protected dogs from challenge with a circulating strain of rabies virus for more than a year. This is the first study to demonstrate that immunization with an NDV-vectored vaccine can induce long-lasting, systemic protective immunity against rabies.
Journal of Virology 06/2011; 85(16):8241-52. · 5.40 Impact Factor
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ABSTRACT: To identify an agent with specific activity against human lens epithelial cells (HLECs), we confirmed the presence of glucocorticoid receptors (GRs) and GR-α genes and evaluated whether GRs have a relationship with the apoptotic process in cultured HLECs. We also determined whether the inhibitor RU486 could rescue the cells from apoptosis when the HLECs were exposed to dexamethasone (Dex), a steroid, in 4 concentrations for 4 periods, or were co-treated with the antagonist RU486. We found that Dex, which has been used as a medical agent for a long time, resulted in increased expression of GRE-luciferase, the GR-α gene and GR-protein and, in contrast, decreased the viability of HLECs. The expression of Bax protein was increased in an earlier stage in contrast to the expression of Bcl-2 protein, which was increased in a later stage. Caspase-3 activity was significantly increased under lower concentrations of Dex in the last stage. The nuclear morphology of HLECs showed an obvious apoptotic phenomenon under greater concentrations of Dex in the last stage. However, RU486, a GR antagonist, could partially inhibit GR and Bax expressions and the expression of caspase-3 was increased so that there was not a decrease in the ratio of apoptotic cells and an increase in the viability of HLECs. Our data showed that GRs had a partial relationship to the apoptotic process of HLECs when exposed to Dex and RU486 did not rescue the cells fully. Because of its toxicity, RU486 did not provide a therapeutic benefit in a glucocorticoid induced cataract (GIC) for the in vitro model, however, its activity and pathway targeting should still be studied further with appropriate drug combinations.
Molecular BioSystems 03/2011; 7(6):1926-37. · 3.53 Impact Factor
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Weiye Chen,
Wenyan Cao,
Huijun Zhao,
Qianqian Hu,
Linmao Qu,
Sen Hu, Jinying Ge,
Zhiyuan Wen,
Xijun Wang,
Haobo Li,
Kehe Huang,
Zhigao Bu
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ABSTRACT: A CHO cell clone (CHO-PoIFN-β) with stable porcine IFN-β expression under control of CMV promoter was selected under G418 pressure. In a 25cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-β reached 2.3×10(6) IU/ml. This cells clone maintained stable expression for at least 20 generations even in the absence of G418 selection pressure. The expressed recombinant PoIFN-β could induce the expression of porcine Mx protein in PK15 cells, and activate the chicken Mx promoter-controlled luciferase reporter gene expression, confirming that the recombinant PoIFN-β has the biological activity of natural porcine type-I interferon. In addition, the recombinant PoIFN-β fully protected PK15 cells against 1000 TCID(50) of porcine transmissible gastroenteritis virus and pseudo-rabies virus infection, demonstrating its high potential in therapeutic applications. This is the first report of establishing a mammalian cell line with stable expression of porcine IFN-β.
Cytokine 03/2011; 54(3):324-9. · 3.02 Impact Factor
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Songhua Shan,
Yongping Jiang,
Zhigao Bu,
Trevor Ellis,
Xianying Zeng,
John Edwards,
Guobin Tian,
Yanbing Li, Jinying Ge,
Hualan Chen,
Stan Fenwick
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ABSTRACT: A low-pathogenicity avian influenza H6N2 virus was used to investigate approaches to improve DNA vaccine efficacy. The viral hemagglutinin (HA) gene or its chicken biased HA gene, incorporating a Kozak sequence, was cloned into a pCAGGS vector to produce the pCAG-HAk and pCAG-optiHAk constructs. Following two intramuscular injections, the seroconversion rate in vaccinated chickens with 10, 100 or 300 μg pCAG-HAk were 87.5%, 75% and 75%, respectively. The profile of H6 hemagglutination inhibition (HI) antibodies induced by different doses of pCAG-HAk during the 8-week study period was similar. The HI titer rose significantly in the three different dose groups following the booster and reached a plateau 2-3 weeks post-booster. In a single dose vaccination group with 100 μg pCAG-HAk, a maximum seroconversion rate reached 53.3% at 5 weeks post-vaccination. The earliest time of seroconversion appeared two weeks after DNA immunization. Following two electroporation (EP) vaccinations with 100 μg pCAG-HAk, all birds seroconverted and the HI antibody titers were significantly higher than those using intramuscular immunization, suggesting that EP was more efficient than intramuscular delivery of the DNA vaccines. In comparison, chickens immunized with 10 or 100 μg pCAG-optiHAk showed 37.5% and 87.5% seroconversion rates, respectively, at 3 weeks following the booster. The pCAG-HAk was not significantly different from the pCAG-optiHAk in either the seroconversion rate or H6 HI titer, suggesting that the codon-optimized HA DNA vaccine did not achieve significantly better immunogenicity than the pCAG-HAk vaccine.
Journal of virological methods 02/2011; 173(2):220-6. · 2.13 Impact Factor
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ABSTRACT: The molecular mechanisms associated with rabies virus (RV) virulence are not fully understood. In this study, the RV Flury low-egg-passage (LEP) and high-egg-passage (HEP) strains were used as models to explore the attenuation mechanism of RV. The results of our studies confirmed that the R333Q mutation in the glycoprotein (G(R333Q)) is crucial for the attenuation of Flury RV in mice. The R333Q mutation is stably maintained in the HEP genome background but not in the LEP genome background during replication in mouse brain tissue or cell culture. Further investigation using chimeric viruses revealed that the polymerase L gene determines the genetic stability of the G(R333Q) mutation during replication. Moreover, a recombinant RV containing the LEP G protein with the R333Q mutation and the HEP L gene showed significant attenuation, genetic stability, enhancement of apoptosis, and immunogenicity. These results indicate that attenuation of the RV Flury strain results from the coevolution of G and L elements and provide important information for the generation of safer and more effective modified live rabies vaccine.
Journal of Virology 09/2010; 84(17):8926-36. · 5.40 Impact Factor
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Yanbing Li,
Jianzhong Shi,
Gongxun Zhong,
Guohua Deng,
Guobin Tian, Jinying Ge,
Xianying Zeng,
Jiasheng Song,
Dongming Zhao,
Liling Liu,
Yongping Jiang,
Yuntao Guan,
Zhigao Bu,
Hualan Chen
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ABSTRACT: Despite substantial efforts to control H5N1 avian influenza viruses (AIVs), the viruses have continued to evolve and cause disease outbreaks in poultry and infections in humans. In this report, we analyzed 51 representative H5N1 AIVs isolated from domestic poultry, wild birds, and humans in China during 2004 to 2009, and 21 genotypes were detected based on whole-genome sequences. Twelve genotypes of AIVs in southern China bear similar H5 hemagglutinin (HA) genes (clade 2.3). These AIVs did not display antigenic drift and could be completely protected against by the A/goose/Guangdong/1/96 (GS/GD/1/96)-based oil-adjuvanted killed vaccine and recombinant Newcastle disease virus vaccine, which have been used in China. In addition, antigenically drifted H5N1 viruses, represented by A/chicken/Shanxi/2/06 (CK/SX/2/06), were detected in chickens from several provinces in northern China. The CK/SX/2/06-like viruses are reassortants with newly emerged HA, NA, and PB1 genes that could not be protected against by the GS/GD/1/96-based vaccines. These viruses also reacted poorly with antisera generated from clade 2.2 and 2.3 viruses. The majority of the viruses isolated from southern China were lethal in mice and ducks, while the CK/SX/2/06-like viruses caused mild disease in mice and could not replicate in ducks. Our results demonstrate that the H5N1 AIVs circulating in nature have complex biological characteristics and pose a continued challenge for disease control and pandemic preparedness.
Journal of Virology 09/2010; 84(17):8389-97. · 5.40 Impact Factor
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ABSTRACT: In order to ensure the biosafety of the IFN-gamma antiviral activity assay, we used a replication-deficient VSV carrying GFP as an interferon sensitive indicator virus (VSVdeltaG*G). The antiviral activities of porcine IFN-gamma expressed in Escherichia coli and in baculovirus on MDBK cells were assessed. The results showed that the antiviral activity of porcine IFN-gamma expressed in baculovirus could reach 10(5) IU/mL, while the porcine IFN-gamma expressed in E. coli showed some antiviral activity (32 IU/mL) after refolding. The results of the VSVdeltaG*G-based antiviral assay were almost identical to that of the VSV*GFP-based assay, suggesting it is highly feasible to use VSVdeltaG*G as a substitute for VSV*GFP, making assays for IFN-gamma antiviral activity safer and more accurate.
Sheng wu gong cheng xue bao = Chinese journal of biotechnology 04/2010; 26(4):439-47.
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ABSTRACT: Infection with H9 avian influenza virus (AIV) and Newcastle disease virus (NDV) are two important causes of egg drop in layer and breeder poultry, leading to severe economic loss in the industry. Currently in China, inactivated H9 AIV vaccine and live attenuated NDV vaccine have to be repeatedly administered to prevent egg drop in layer animals. Using reverse genetics, we constructed a recombinant NDV expressing an H9 AIV hemagglutinin (HA) from an H9N2 field isolate, A/Chicken/ Shandong/2/2007. The HA gene was inserted into the intergenic region between the phosphoprotein (P) and matrix (M) genes of the LaSota NDV vaccine strain. The recombinant virus stably expressing the HA gene, rL-H9, was found to be innocuous after intracerebral inoculation of 1-day-old chickens. A single dose of 10(6) 50% egg infectious dose of the recombinant virus intranasally inoculated into chickens induced high levels of NDV- and AIV H9-specific hemagglutination-inhibition antibody. Complete protection from clinical disease and mortality against challenge with a lethal dose of velogenic NDV was observed in chickens and 90% of chickens were protected from clinical disease, mortality, and virus shedding against challenge with homologous H9N2 AIV. Our results suggest that recombinant NDV is suitable as a potential bivalent live attenuated vaccine against both NDV and H9 AIV infection in poultry.
Avian Diseases 03/2010; 54(1 Suppl):294-6. · 1.46 Impact Factor
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ABSTRACT: Oxidative damage induced by H(2)O(2) treatment can irreversibly damage the lens epithelium, resulting in cell death and cataract. Whether the effects of oxidative stress could be attenuated in cultured human lens epithelial cells by incubation with resveratrol (RES) is still unknown. In the present study, we examined the function of resveratrol in protecting human lens epithelial B-3 (HLEB-3) cells against H(2)O(2) induced cell death and cell apoptosis, its role in reducing H(2)O(2) induced intracellular reactive oxygen species (ROS) accumulation, and investigated the mechanism by which resveratrol underlies the effect.
HLEB-3 cells, a human lens epithelial cell line, were exposed to 100 muM H(2)O(2) with or without RES pre-treatment at different concentrations for different time duration. Cell viabilities were monitored by 4-[3-[4-iodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] (WST-1) assay. The apoptosis rate and ROS generation were detected by flow cytometric analysis. Expression levels of superoxide dismutases-1 (SOD-1), catalase, and heme oxygenase-1 (HO-1) proteins were measured by western-blotting analysis. p38 and c-jun N terminal kinase (JNK) activation was also evaluated by western-blotting analysis.
Resveratrol clearly reduced H(2)O(2) induced cell apoptosis and ROS accumulation; protected HLEB-3 cells from H(2)O(2) induced oxidative damage, and increased the expression levels of SOD-1, catalase, and HO-1. Further studies showed that RES also inhibited H(2)O(2) induced p38 and JNK phosphorylation.
These findings suggested that RES protected HLEB-3 cells from H(2)O(2) induced oxidative damage, presumably by inducing three antioxidative enzymes including catalase, SOD-1, and HO-1.
Molecular vision 01/2010; 16:1467-74. · 2.20 Impact Factor
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ABSTRACT: To establish a reverse genetic system of rabies virus for producing safe and efficient rabies vaccine.
By reverse genetic and molecular cloning technique, we established two rabies virus rescue systems including (1) help plasmids expressing N, P and L protein, and (2) CMV/T7 and T7 promoter.
Wild-type rERA-VC was rescued by both systems, and had the same growth kinetics as parental virus. The third generation virus could grow to high titer.
The established reverse genetic system for rescuing wild-type rERA-VC provides the possibility of producing safe and efficient rabies virus vaccines.
ACTA MICROBIOLOGICA SINICA 07/2009; 49(7):949-54.
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ABSTRACT: The hemagglutinin-neuraminidase (HN) and fusion protein of Newcastle disease virus (NDV) plays a crucial role in the process of budding and infection. To understand the exact contribution of the HN gene to NDV pathogenecity, a reverse genetics system was developed using the lentogenic NDV LaSota strain .
the HN genes of an avirulent recombinant NDV strain (rLaSota) was replaced by an HN gene from Mukteswar mesogenic NDV strain by reverse genetics. Furthermore, the F gene of rL-MuHN was replaced with that of Mukteswar strain, resulting the double genes replaced chmeric virus, rL-MuFHN.
Although the rescued chimeric virus (rL-MuHN) did not show significant increase in ability of hemadsorption, the intracerebral pathogenicity index test (ICPI = 0) in chickens and mean death time for eggs (MDT > or = 90 h). rL-MuHN kept the low pathogenicity similar to its parent rLaSota strain. Compared to single gene replaced rL-MuHN, rL-MuFHN induced stronger cell fusion and showed a mild increase in ICPI (from 0 to 0.59) and no significant change in MDT (> or = 90 h). rL-MuFHN showed much lower pathogenicity than that of Mukteswar (ICPI. = 1.32 and MDT = 46, respectively). A HN gene exchange alone within the context of the NDV rLaSota backbone failed to increase virus virulence from unvelogenic to mesogenic pathotype.
These results indicated that the virulence of NDV is determined multigenically. The heterotypic HN and F pairs were not equally effective in virus pathogenicity. The HN gene derivated from mesogenic strain dose not alter the lentogenic property of NDV LaSota strain. NDV can be manipulated by gene replacement in the future for use as a vaccine candidate.
ACTA MICROBIOLOGICA SINICA 05/2008; 48(5):638-43.
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ABSTRACT: Newcastle disease virus (NDV) cause a highly contagious and economically loss in poultry. The amino acid sequence at the protease cleavage site of the fusion (F) protein has been postulated as a major determinant of NDV virulence. In this study, we have examined the role of F protein cleavage site sequence in NDV virulence by use reverse genetics technology. The sequence G-R-Q-G-R-L present at the cleavage site of the F protein of avirulent strain LaSota was mutated to R-R-Q-R-R-F or R-R-Q-R-R-L. The resultant mutated rL-FmF and rL-FmL virus were evaluated by mean death test (MDT) in eggs, intracerebral pathogenicity index (ICPI) and intravenous pathogenicity index (IVPI) tests in chickens. The results showed that the modification of the F protein cleavage site resulted in a dramatic in crease in virulence from an ICPI value of 0.36 for LaSota to a value of 1.18 for rL-FmF and 1.06 for rL-FmL, respectively. In addition, the mutational viruses showed increase MDT and identical IVPI values to parent virus, the virulence of rescued viruses was greatly enhanced by the amino acid replacements, whether the amino acid on the N terminus of F2 was F or L. On this base, we constructed another two NDVs that expressed a H5 subtype avian influenza virus hemagglutinin (rL-FmF-HA) and green fluorescent protein (rL-FmF-EGFP). the ICPI value of recombinant virus rL-FmF-HA and rL-FmF-EGFP were 0.67 and 1.10, respectively lower than that of rL-FmF. the IVPI of both recombinant viruses still keep 0.00. The values of MDT for rL-FmF-HA and rL-FmF-EGFP were 117h and 101h, greater than 90h. thus, Introduction of a foreign gene into NDV genome resulted in growth retardation and attenuation, the decrease degree depended on the nature of foreign protein expressed. These results also indicate that cleavability of the F0 protein is an important determinant for virulence of NDV. NDV can be manipulated in the future for use as a vaccine vector.
ACTA MICROBIOLOGICA SINICA 04/2008; 48(3):362-8.
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Yongping Jiang,
Kangzhen Yu,
Hongbo Zhang,
Pingjing Zhang,
Chenjun Li,
Guobin Tian,
Yanbing Li,
Xijun Wang, Jinying Ge,
Zhigao Bu,
Hualan Chen
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ABSTRACT: H5N1 influenza viruses have caused significant disease and deaths in various parts of the world in several species, including humans. Vaccination combined with culling can provide an attractive method for outbreak containment. Using synthesized oligos and overlapping extension PCR techniques, we constructed an H5 HA gene, optiHA, containing chicken biased codons based on the HA amino acid sequence of the highly pathogenic H5N1 virus A/goose/Guangdong/1/96 (GS/GD/96). The optiHA and wild-type HA genes were inserted into plasmids pCI or pCAGGS, and designated as pCIoptiHA, pCAGGoptiHA, pCIHA and pCAGGHA, respectively. To evaluate vaccine efficacy, groups of 3-week-old specific pathogen free (SPF) chickens were intramuscularly injected with the four plasmids. Sera were collected on a weekly basis post-vaccination (p.v.) for hemagglutination inhibition (HI) assays and neutralization (NT) antibody detection. All chickens receiving pCAGGoptiHA and pCAGGHA developed high levels of HI and NT antibodies at 3 weeks p.v., and were completely protected from lethal H5 virus challenge, while only partial protection was induced by inoculation with the other two plasmids. A second experiment was conducted to evaluate if a lower dose of the pCAGGoptiHA vaccine could be effective, results indicated that two doses of 10 microg of pCAGGoptiHA could induce complete protection in chickens against H5 lethal virus challenge. Based on our results, we conclude that construction optimization could dramatically increase the H5 HA gene DNA vaccine efficacy in chickens, and therefore, greatly decrease the dose necessary for inducing complete protection in chickens.
Antiviral Research 10/2007; 75(3):234-41. · 4.30 Impact Factor
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Jinying Ge,
Guohua Deng,
Zhiyuan Wen,
Guobing Tian,
Yong Wang,
Jianzhong Shi,
Xijun Wang,
Yanbing Li,
Sen Hu,
Yongping Jiang,
Chinglai Yang,
Kangzhen Yu,
Zhigao Bu,
Hualan Chen
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ABSTRACT: H5N1 highly pathogenic avian influenza virus (HPAIV) has continued to spread and poses a significant threat to both animal and human health. Current influenza vaccine strategies have limitations that prevent their effective use for widespread inoculation of animals in the field. Vaccine strains of Newcastle disease virus (NDV), however, have been used successfully to easily vaccinate large numbers of animals. In this study, we used reverse genetics to construct a NDV that expressed an H5 subtype avian influenza virus (AIV) hemagglutinin (HA). Both a wild-type and a mutated HA open reading frame (ORF) from the HPAIV wild bird isolate, A/Bar-headed goose/Qinghai/3/2005 (H5N1), were inserted into the intergenic region between the P and M genes of the LaSota NDV vaccine strain. The recombinant viruses stably expressing the wild-type and mutant HA genes were found to be innocuous after intracerebral inoculation of 1-day-old chickens. A single dose of the recombinant viruses in chickens induced both NDV- and AIV H5-specific antibodies and completely protected chickens from challenge with a lethal dose of both velogenic NDV and homologous and heterologous H5N1 HPAIV. In addition, BALB/c mice immunized with the recombinant NDV-based vaccine produced H5 AIV-specific antibodies and were completely protected from homologous and heterologous lethal virus challenge. Our results indicate that recombinant NDV is suitable as a bivalent live attenuated vaccine against both NDV and AIV infection in poultry. The recombinant NDV vaccine may also have potential use in high-risk human individuals to control the pandemic spread of lethal avian influenza.
Journal of Virology 02/2007; 81(1):150-8. · 5.40 Impact Factor
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ABSTRACT: We investigated the antibody response of DNA immunization with two mammalian codon optimized envelope glycoprotein genes, F and G, of Nipah virus in a mouse model. The results indicated that G gene immunization elicited more significant specific serum IgG response and neutralization antibody response than F gene did, suggesting that the G gene DNA immunization is a potential vaccine strategy against Nipah virus.
Annals of the New York Academy of Sciences 11/2006; 1081:243-5. · 3.15 Impact Factor
