The ability of Hepatitis B surface antigen DNA vaccine to elicit cell-mediated immune responses, but not antibody responses, was affected by the deglysosylation of S antigen

Xing, Yiping; Huang, Zhibin; Lin, Yan; Li, Jun; Chou, Te-hui W.; Lu, Shan; Wang, Shixia

doi:10.1016/j.vaccine.2008.03.072

Cited by 12 publications

(21 citation statements)

References 43 publications

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“…The expression of the MHBs-encoding DNA vaccine was confirmed by western blot analysis using the polyclonal mouse immune sera, elicited by an MHBs-encoding DNA vaccine (Xing et al 2008). The two glycosylation sites of MHBs are schematically shown in Fig.…”

Section: In Vitro Expression Of Mutated Mhbs-encoding Dna Vaccinesmentioning

confidence: 97%

“…Blocking was performed with 0.1% I-Block (Tropix, Bedford, MA). MHBsspecific mouse sera produced in a previous study (Xing et al 2008) were used as the detecting antibody at 1:500 dilution with incubation for 45 min. Subsequently, membranes were washed with blocking buffer and then reacted with alkaline phosphatase (AP)-conjugated goat anti-rabbit (Tropix) at 1:5000 dilution.…”

Section: Western Blot Analysismentioning

confidence: 99%

“…Serum samples were collected prior to the first immunization and two weeks after each immunization to measure HBs-specific antibody responses. Spleens were removed and single-cell suspensions of splenocytes were prepared, as previously described (Xing et al 2008). This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.…”

Section: Mhbs-n4-7 Mqwqstnftqmentioning

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%

“…MHBs possesses two N-linked glycosylation sites: Asn4 in the pre-S2 domain and Asn146 in the S domain. A previous analysis showed that deglycosylation at N-linked glycosylation site (Asn146) in the S domain resulted in a significant decrease in HBs-specific cell-mediated immune responses (Xing et al 2008), and there is evidence that N-linked glycosylation (Asn4) in the pre-S2 domain plays an essential role in the secretion of MHBs and formation of HBV particles (Block et al 1994). …”

Section: Introductionmentioning

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: In Vitro Expression Of Mutated Mhbs-encoding Dna Vaccinesmentioning

confidence: 97%

Section: Western Blot Analysismentioning

confidence: 99%

Section: Mhbs-n4-7 Mqwqstnftqmentioning

confidence: 99%

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

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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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DNA Immunization

Wang

2013

CP Microbiology

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DNA immunization was discovered in early 1990s, and its use has been expanded from vaccine studies to a broader range of biomedical research areas, such as the generation of high-quality polyclonal and monoclonal antibodies as research reagents. In this unit, three common DNA immunization methods are described: needle injection, electroporation, and gene gun. In addition, several common considerations related to DNA immunization are discussed.

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rpoS involved in immune response of Macrobrachium nipponens to Vibrio mimicus infection

Jiang,

Qin,

Chen

et al. 2024

Fish & Shellfish Immunology

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The ability of Hepatitis B surface antigen DNA vaccine to elicit cell-mediated immune responses, but not antibody responses, was affected by the deglysosylation of S antigen

Cited by 12 publications

References 43 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

DNA Immunization

rpoS involved in immune response of Macrobrachium nipponens to Vibrio mimicus infection

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