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ABSTRACT: INTRODUCTION: Integration-deficient lentiviral vectors (IDLVs) are a promising platform for immunisation to elicit both humoral immunity and cellular mediated immunity (CMI). Here, we compared the specific immunity in mice immunised via different regimens (homologous and cocktail) with IDLV-based HCV pseudoparticles (HCVpps) carrying pseudotyped glycoproteins E1E2 and bearing the HCV NS3 gene. Humoral and cell-mediated immune responses were also evaluated after IDLV-HCVpp immunisation combined with heterologous rAd5-CE1E2 priming protocols. Sera from the mice effectively elicited anti-E1, -E2, and -NS3 antibody responses, and neutralised various HCVpp subtypes (1a, 1b, 2a, 3a and 5a). No significant CMI was detected in the groups immunised with IDLV-based HCVpps. In contrast, the combination of rAd5-CE1E2 priming and IDLV-based HCVpp boosting induced significant CMI against multiple antigens (E1, E2, and NS3). CONCLUSION: IDLV-based HCVpps are a promising vaccination platform and the combination of rAd5-CE1E2 and IDLV-based HCVpp prime-boost strategy should be further explored for the development of a cross-protective HCV vaccine.
PLoS ONE 01/2013; 8(4):e62684. · 4.09 Impact Factor
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ABSTRACT: An effective vaccine and new therapeutic methods for hepatitis C virus (HCV) are needed, and a potent HCV vaccine must induce robust and sustained cellular-mediated immunity (CMI). Research has indicated that adenoviral and vaccinia vectors may have the ability to elicit strong B and T cell immune responses to target antigens.
A recombinant replication-defective adenovirus serotype 5 (rAd5) vector, rAd5-CE1E2, and a recombinant Tian Tan vaccinia vector, rTTV-CE1E2, were constructed to express the HCV CE1E2 gene (1-746 amino acid HCV 1b subtype). Mice were prime-immunised with rAd5-CE1E2 delivered via intramuscular injection (i.m.), intranasal injection (i.n.), or intradermal injection (i.d.) and boosted using a different combination of injection routes. CMI was evaluated via IFN-γ ELISPOT and ICS 2 weeks after immunisation, or 16 weeks after boost for long-term responses. The humoral response was analysed by ELISA. With the exception of priming by i.n. injection, a robust CMI response against multiple HCV antigens (core, E1, E2) was elicited and remained at a high level for a long period (16 weeks post-vaccination) in mice. However, i.n. priming elicited the highest anti-core antibody levels. Priming with i.d. rAd5-CE1E2 and boosting with i.d. rTTV-CE1E2 carried out simultaneously enhanced CMI and the humoral immune response, compared to the homologous rAd5-CE1E2 immune groups. All regimens demonstrated equivalent cross-protective potency in a heterologous surrogate challenge assay based on a recombinant HCV (JFH1, 2a) vaccinia virus.
Our data suggest that a rAd5-CE1E2-based HCV vaccine would be capable of eliciting an effective immune response and cross-protection. These findings have important implications for the development of T cell-based HCV vaccine candidates.
Virology Journal 11/2011; 8:506. · 2.34 Impact Factor
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ABSTRACT: To develop a novel, effective HBV therapeutic vaccine, we constructed two HBV DNA immunogens that contained PreS1, HBSS1, and HBCS1. Several delivery methods, such as intramuscular (i.m.) injection, intramuscular injection plus electroporation (i.m.-EP), and intradermal injection plus electroporation (i.d.-EP) were used in a murine model to analyze and compare the immune responses that were induced by the DNA immunogens. We found that i.d.-EP accelerated specific antibody seroconversion and produced high antibody (anti-PreS1, anti-S, and anti-C antibody) titers after HBSS1 and HBCS1 immunization. Combining the HBSS1 and HBCS1 DNA immunogens with i.d.-EP produced the strongest multiantigen (PreS1, S, and C)-specific cellular immune response and the highest specific PreS1 antibody levels. The results indicated that DNA immunization using HBSS1 and HBCS1 might be an ideal candidate, with its ability to elicit robust B and T cell immune responses against multiantigen when combined with optimized delivery technology. The present study provides a basis for the design and rational application of a novel HBV DNA vaccine.
Clinical and vaccine immunology: CVI 09/2011; 18(11):1789-95. · 2.37 Impact Factor
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ABSTRACT: A large number of researches focused on glycoproteins E1 and E2 of hepatitis C virus (HCV) aimed at the development of anti-HCV vaccines and inhibitors. Enhancement of E1/E2 expression and secretion is critical for the characterization of these glycoproteins and thus for subunit vaccine development. In this study, we designed and synthesized three signal peptide sequences based on online programs SignalP, TargetP, and PSORT, then removed and replaced the signal peptide preceding E1/E2 by overlapping the polymerase chain reaction method. We assessed the effect of this alteration on E1/E2 expression and secretion in mammalian cells, using western blot analysis, dot blot, and Galanthus nivalis agglutinin lectin capture enzyme immunoassay. Replacing the peptides preceding E1 and E2 with the signal peptides of the tissue plasminogen activator and Gaussia luciferase resulted in maximum enhancement of E1/E2 expression and secretion of E1 in mammalian cells, without altering glycosylation. Such an advance would help to facilitate both the research of E1/E2 biology and the development of an effective HCV subunit vaccine. The strategy used in this study could be applied to the expression and production of other glycoproteins in mammalian cell line-based systems.
Acta Biochimica et Biophysica Sinica 02/2011; 43(2):96-102. · 1.38 Impact Factor
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ABSTRACT: To develop novel and effective HBV therapeutic vaccine, we constructed an expression vector, pVRC-HBSS1, in which PreS1 (21-47aa) coding gene fused to the C-terminal of the S (1-223 aa) coding gene of HBV, and prepared the protein particle vaccine HBSS1 that consist of S and PreS1 fusion antigen derived from CHO system. We immunized mice by priming three times with DNA vaccine via different methods (i.e., intramuscular injection, intradermal injection with electroporation), then boosting once with protein particle vaccine. We analyzed the immune response among various vaccination groups. The higher level of S or PreS1 specific antibodies was detected in the group via intradermal injection with electroporation, compared with that of direct intramuscular injection. We further found that the specific cellular immune responses (IFN-gamma ELISpot analysis) in the group priming with DNA vaccines and boosting with protein subunit vaccine particles, was significantly higher than that of the DNA or protein particle subunit alone. Moreover, combination vaccination priming with intradermal injection DNA via electroporation and boosting with protein particle induced the strongest cellular immune response. These results provide a basis for rational design and application of the novel HBV therapeutic vaccine.
Sheng wu gong cheng xue bao = Chinese journal of biotechnology 01/2011; 27(1):95-100.
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ABSTRACT: To develop a new Hepatitis C virus (HCV) DNA vaccine ,We explored strategies for optimizing the immunogenicity of HCV DNA vaccine in fusion with dendritic cell-targeting molecules (scDEC, a single-chain antibody against the murine DC cell surface molecule DEC205). We constructed the DNA vaccine plasmids expressing the HCV non-structural protein NS3 alone or in combination with DEC205 as a fusion protein and identified the expression of the molecules of interest by transient transfection of 293 cells with the resultant DNA vaccine plasmids. Then BALB/C mice were immunized with these plasmids by the injection in combination with electroporation. The NS3-specific IgG antibody(ELISA) and cellular immunity (IFN-gamma ELISPOT) were analyzed post twice immunization. Our results showed that: the single-chain antibody against DEC205 fused with vaccine antigen NS3 significantly enhanced the immunogenicity of new HCV DNA vaccine, the intradermal injection in combination with electroporation using caliper electrodes resulted in most robust NS3-specific antibody and T cell immune response. In conclusion,immune response for the HCV NS3 protein-encoding DNA vaccine was enhanced significantly when targeting antigen NS3 to DCs by scDEC. The present strategy could merit further study in the context of other prophylactic and therapeutic DNA based vaccines.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 01/2011; 27(1):44-9.
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Wen Wang,
Hong Chen,
Wen-jie Tan,
Yao Deng,
Min Wang,
Yuan Liu,
Xiao Yin,
Ke Zhang, Jie Guan,
Jian-fang Zhou,
Yue-long Shu,
Li Ruan
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ABSTRACT: This study aims to develop inexpensive and effective experimental vaccines against highly pathogenic H5N1 Avian Influenza (HPAI) virus and to optimize their immunization programs. To this end, we first synthesized the codon-optimized hemagglutinin gene (HAop) and neuraminidase gene (NAop), both of which were derived from a H5N1 virus (Anhui strain), and constructed successfully the DNA vaccines containing a single cistronic construct (HAwt, HAop, or NAop) or a bicistronic construct (HAop/M2 or NAop/M1) of H5N1 influenza virus origin. Their expression was confirmed by indirect immunofluorescent assay (IFA) and Western blotting. Then twice vaccination of mice with the DNA vaccines by injection intramuscularly or in vivo electroporation (EP) via two different routes was evaluated and analyzed by hemagglutination inhibition (HI) assay, NA-specific antibody detection, micro-neutralizing antibody test and IFN-gamma ELISpot assay. Our results showed that the DNA vaccines with coden-optimized HAop and NAop constructs could quickly elicit a strong immune response by in vivo EP, especially the cellular immune response against HA and NA; the in vivo EP via intradermal route induced stronger humoral immune responses than those via intramuscular route. Our findings will pave a way for further development of novel DNA-based H5N1 vaccine and for the optimization of the immunization programs of DNA vaccine.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 05/2010; 26(3):170-5.
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ABSTRACT: To rational design HBV therapeutic vaccine candidate and evaluate their specific immunity to HBV in mice.
Based on our previous data of HBV protein vaccine consisting of S-PreS1 fusion particle. We first construct a novel DNA vaccine candidate, pVRC-HBSS1, which consisting of S (aa: 1-223) and PreS1 (aa: 21-47) fuse gene,then confirm the expression of the DNA vaccine by Western blotting, and followed by vaccination using prime boost strategy, ie, Intradermal injection of DNA vaccine with gene electroporation (EP) in BALB/c mice after twice injection of different HBSS1 protein vaccines (combination with different adjuvants). The immune response was measured by ELISA and IFN-gamma ELISPOT.
The novel DNA vaccine candidate could effectively express in vitro, boost with single intradermal injection of HBV DNA vaccine via EP can significantly enhance the surface antigen (S)-specific cellular immune responses (IFN-gamma ELISpot analysis) and PreS1-specific antibody levels, especially in the group primed with protein vaccine in combination with alum adjuvant.
Boost with the novel HBV DNA vaccine followed prime with HBV protein vaccine could induced a higher anti-HBV T cell response in mice than vaccination with the HBSS1 particle-like protein vaccine only. This prime-boost vaccination may serve as a promising way to develop and optimize the novel HBV therapeutic vaccine.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 04/2010; 24(2):94-7.
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ABSTRACT: We previous reported the development of novel hepatitis B virus(HBV) vaccine containing the surface antigen(S) plus PreS1 fusion derived from Chinese hamster ovary (CHO) cells system. In this study, we analyzed the impact of different adjuvants on immunogenicity of the HBV particle vaccine in Balb/C mice, including alum alone, CpG oligodeoxynucleotides (CpG-ODN) alone and CpG-ODN in combination with alum adjuvant. We first detected the antigen specific humoral response in mice, including total IgG antibody and IgG subtyping. Then, we characterized the specific cell-mediated immune (CMI) response by detection of gamma-interferon secreting splenocytes after stimulating with S or PreS1 peptide pool. Our results showed that: CpG-ODN adjuvanted vaccine could rapidly induce higher level of anti-PreS 1 and anti-S antibodies, and a higher ratio of IgG2a/IgG1 antibody than that of alum adjuvanted vaccine. At the same time, CpG-ODN adjuvanted vaccine induced robust antigen-specific cellular immune responses in mice, which was superior to that of alum adjuvanted vaccine and CpG-ODN in combination with alum adjuvanted vaccine; however, the vaccine candidate with CpG-ODN in combination with alum adjuvant induced highest anti-S antibody and mixed IgG subclasses in mice after twice immunization. There exists dominant HBV CMI epitopes in the N-terminal of S antigen. These results provided important evidence that CpG-ODN adjuvanted HBSS1 particles vaccine may serve as a novel candidate in the development of new preventive and therapeutic agents against hepatitis B infection.
Sheng wu gong cheng xue bao = Chinese journal of biotechnology 01/2010; 26(1):74-8.
