Profiles of B and T cell immune responses elicited by different forms of the hepatitis B virus surface antigen

Shen, Meilong; Wang, Shixia; Gao, Ge; Xing, Yiping; Ma, Xiuwen; Huang, Zhibin; Lu, Shan

doi:10.1016/j.vaccine.2010.08.081

Cited by 13 publications

(20 citation statements)

References 24 publications

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“…Our work further extends this finding and indicates that any gene-based vaccines or therapies based on MHBs should consider the impact of the N-linked glycosylation site at position 4 in the pre-S2 domain. HBV surface proteins possess a complicated design and structure, with three different forms (SHBs, MHBs, and LHBs), each of which showed a distinct pattern of immunogenicity when tested as DNA vaccines, as we reported previously (Shen et al 2010;Ge et al 2012). Our data also provide new information showing that this N-linked glycosylation site can remain functional after relocation to other positions within the pre-S2 domain.…”

Section: Discussionsupporting

confidence: 73%

“…Anti-MHBs antibodies in immunized animal sera were evaluated by enzyme-linked immunosorbent assay (ELISA), as previously described (Shen et al 2010;Ge et al 2012). Both temporal and peaklevel MHBs-specific antibodies were determined.…”

Section: Detection Of Anti-mhbs Antibody Responses In Mouse Immune Seramentioning

confidence: 99%

“…Gamma interferon (IFN-γ) ELISPOT assays were performed to detect MHBs peptide-specific T cell responses in mouse splenocytes, as described previously (Xing et al 2008;Shen et al 2010;Ge et al 2012;Chuai et al 2013). A mouse IFN-γ kit (U-CyTech Biosciences, the Netherlands) was used to detect MHBs-specific IFN-γ responses, according to the manufacturers' directions.…”

Section: Analysis Of Mhbs-specific T Cell Responses By Enzyme-linked mentioning

confidence: 99%

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N-Linked Glycosylation at an Appropriate Position in the Pre-S2 Domain Is Critical for Cellular and Humoral Immunity against Middle HBV Surface Antigen

Líu

Wang

Jia

et al. 2015

Tohoku J. Exp. Med.

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Infection with hepatitis B virus (HBV) remains a worldwide health problem, and DNA-based vaccines against HBV have been tested for therapeutic applications. HBV possesses three envelope lipoproteins that are translated from a single reading-frame: large, middle, and small HBV surface antigens. Among these envelope proteins, the middle HBV surface antigen (MHBs) contains a constitutive N-linked glycosylation site at position 4 (Asn4) in the amino-terminal portion (MQWNSTTFHQ) of pre-S2 domain. Asn4 (shown in bold) is essential for secretion of viral particles and conserved among all serotypes of HBV, but its influence on the immunogenicity of MHBs remains unknown. Here, we constructed four MHBs genes carrying mutations, underlined, in the amino-terminal portion of pre-S2 domain. One mutant protein contains Q at position 4 (MQWQSTTFHQ). In addition, each of three mutant MHBs proteins contains a N-linked glycosylation site (N-X-S/T), relocated to position 5 (MQWQNTTFHQ), 6 (MQWQSNTSHQ) or 7 (MQWQSTNFTQ) in pre-S2 domain. The expression and immunogenic properties of mutant DNA vaccines were examined in 293T human renal epithelial cells and in BALB/c mice, respectively. We showed that Asn4 was critical for secretion and immunogenicity of MHBs. Moreover, the MHBs protein that carries a N-linked glycosylation site at position 5 or 7 retained the properties similar to wild-type MHBs. In contrast, the secretion-defective mutant protein carrying Asn at position 6 induced only marginal humoral and cellular immune responses in mice, despite the N-linked glycosylation. In conclusion, N-linked glycosylation at an appropriate position in pre-S2 domain is an essential requirement for DNA vaccine expressing MHBs.

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Section: Discussionsupporting

confidence: 73%

Section: Detection Of Anti-mhbs Antibody Responses In Mouse Immune Seramentioning

confidence: 99%

Section: Analysis Of Mhbs-specific T Cell Responses By Enzyme-linked mentioning

confidence: 99%

See 1 more Smart Citation

N-Linked Glycosylation at an Appropriate Position in the Pre-S2 Domain Is Critical for Cellular and Humoral Immunity against Middle HBV Surface Antigen

Líu

Wang

Jia

et al. 2015

Tohoku J. Exp. Med.

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“…Only low amounts of S-specific and no PreS1- and PreS2-specific IFN-γ-secreting PBMC were detected. In mice, strong S-specific [32], [33] and negligible PreS1/PreS2-specific [33] cellular responses have been described after immunization with DNA vaccines expressing non-secreted L proteins. The comparably high DNA doses applied in those studies (15 µg [33] or 100 µg DNA [32] for a ∼20 g mouse) as opposed to 333 µg DNA for a ∼20 kg pig used in our study might explain the different level of S-specific cellular immune responses.…”

Section: Discussionmentioning

confidence: 99%

Cationic Lipid-Formulated DNA Vaccine against Hepatitis B Virus: Immunogenicity of MIDGE-Th1 Vectors Encoding Small and Large Surface Antigen in Comparison to a Licensed Protein Vaccine

et al. 2014

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Currently marketed vaccines against hepatitis B virus (HBV) based on the small (S) hepatitis B surface antigen (HBsAg) fail to induce a protective immune response in about 10% of vaccinees. DNA vaccination and the inclusion of PreS1 and PreS2 domains of HBsAg have been reported to represent feasible strategies to improve the efficacy of HBV vaccines. Here, we evaluated the immunogenicity of SAINT-18-formulated MIDGE-Th1 vectors encoding the S or the large (L) protein of HBsAg in mice and pigs. In both animal models, vectors encoding the secretion-competent S protein induced stronger humoral responses than vectors encoding the L protein, which was shown to be retained mainly intracellularly despite the presence of a heterologous secretion signal. In pigs, SAINT-18-formulated MIDGE-Th1 vectors encoding the S protein elicited an immune response of the same magnitude as the licensed protein vaccine Engerix-B, with S protein-specific antibody levels significantly higher than those considered protective in humans, and lasting for at least six months after the third immunization. Thus, our results provide not only the proof of concept for the SAINT-18-formulated MIDGE-Th1 vector approach but also confirm that with a cationic-lipid formulation, a DNA vaccine at a relatively low dose can elicit an immune response similar to a human dose of an aluminum hydroxide-adjuvanted protein vaccine in large animals.

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“…ELISPOT assays were performed on fresh mouse splenocytes as previously described. 57,59,60 Multiscreen Immobilon P membrane White sterile 96-well plates (Millipore) were coated with 5 μg/ml of purified rat anti-mouse IFN-γ IgG1 (clone R4-6A2) in PBS at 4 °C overnight. After the plates were washed three times with PBS, each plate was blocked by the addition of 200 μl of R10 medium per well for 2-3 h at 37 °C.…”

Section: Acknowledgmentsmentioning

confidence: 99%

Post-translational intracellular trafficking determines the type of immune response elicited by DNA vaccines expressing Gag antigen of Human Immunodeficiency Virus Type 1 (HIV-1)

Wallace

West

Rothman

et al. 2013

Human Vaccines & Immunotherapeutics

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In the current study, immune responses induced by Gag DNA vaccines with different designs were evaluated in Balb/C mice. The results demonstrated that the DNA vaccine with the full length wild type gag gene (Wt-Gag) mainly produced Gag antigens intracellularly and induced a higher level of cell-mediated immune (CMI) responses, as measured by IFN-gamma ELISPOT, intracellular cytokine staining (ICS), and cytotoxic T lymphocytes (CTL) assays against a dominant CD8(+) T cell epitope (AMQMLKETI). In contrast, the addition of a tissue plasminogen activator (tPA) leader sequence significantly improved overall Gag protein expression/secretion and Gag-specific antibody responses; however, Gag-specific CMI responses were decreased. The mutation of zinc-finger motif changed Gag protein expression patterns and reduced the ability to generate both CMI and antibody responses against Gag. These findings indicate that the structure and post-translational processing of antigens expressed by DNA vaccines play a critical role in eliciting optimal antibody or CMI responses.

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Profiles of B and T cell immune responses elicited by different forms of the hepatitis B virus surface antigen

Cited by 13 publications

References 24 publications

N-Linked Glycosylation at an Appropriate Position in the Pre-S2 Domain Is Critical for Cellular and Humoral Immunity against Middle HBV Surface Antigen

N-Linked Glycosylation at an Appropriate Position in the Pre-S2 Domain Is Critical for Cellular and Humoral Immunity against Middle HBV Surface Antigen

Cationic Lipid-Formulated DNA Vaccine against Hepatitis B Virus: Immunogenicity of MIDGE-Th1 Vectors Encoding Small and Large Surface Antigen in Comparison to a Licensed Protein Vaccine

Post-translational intracellular trafficking determines the type of immune response elicited by DNA vaccines expressing Gag antigen of Human Immunodeficiency Virus Type 1 (HIV-1)

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