Targeting the vaccinia virus L1 protein to the cell surface enhances production of neutralizing antibodies

Golden, Joseph W.; Josleyn, Matthew; Hooper, Jay W.

doi:10.1016/j.vaccine.2008.04.017

Cited by 36 publications

(40 citation statements)

References 47 publications

(109 reference statements)

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“…Following immunization with IgM-tL1R through electroporation, we observed the induction of a prominent antibody response. At five weeks post-immunization, the antibody titer was approximately 3.0 log titers which were similar to other studies using a modified L1R construct [28,33]. The capacity tL1R-specific antibodies to neutralize vaccinia virus was also confirmed by plaque assay.…”

Section: Discussionsupporting

confidence: 87%

“…In our experiments, the original L1R and codon-optimized tL1R were 50% different in terms of codon usage (data not shown). Previous poxvirus DNA studies demonstrated that signal sequence was able to promote the secretion of antigens and their immune responses [32,33]. These studies indicated that a tissue plasminogen activator signal peptide fused to L1R induced a higher amount of neutralizing antibodies and provided better immunogenicity than unmodified L1R.…”

Section: Discussionmentioning

confidence: 98%

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Immunogenicity and Protective Efficiency in Mice of a Smallpox DNA Vaccine Candidate

Kim¹,

Chang²,

Hur³

et al. 2017

J Bioterror Biodef

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The adverse reactions of the current live smallpox vaccine, and potential use of smallpox as a bioterrorism weapon, have highlighted the need to develop a new effective vaccine for this infectious disease. In the present study, a DNA vaccine vector was produced, which was optimized for expression of the vaccinia virus L1 antigen in a mouse model. Plasmid-encoded IgM-tL1R, which contains a truncated L1R gene fused to an IgM signal sequence, was constructed and expressed under the regulation of an SV40 enhancer. The expressed recombinant tL1 proteins were successfully secreted into the culture media. The DNA vaccine was administered to mice by electroporation, and animals were subsequently challenged with the lethal dose of vaccinia virus. We observed that immunization with IgM-tL1R induced robust neutralizing antibody responses and provided complete protection against a vaccinia virus infection. Isotyping studies revealed a lower IgG1/IgG2a ratio following vaccination with IgM-tL1R, suggesting the stimulation of Th1 immune responses. Our results propose that an optimized DNA vaccine, IgM-tL1R, can be effective in eliciting an anti-vaccinia virus immune response and provide protection against lethal orthopoxvirus challenge.

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Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 98%

Immunogenicity and Protective Efficiency in Mice of a Smallpox DNA Vaccine Candidate

Kim¹,

Chang²,

Hur³

et al. 2017

J Bioterror Biodef

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“…Recently, we further enhanced the protective efficacy of the 4pox vaccine by optimizing the genes for codon usage and for mRNA stability ( Fig. 1 and 2 and data not shown) and enhancing the immunogenicity of the L1 target by targeting the protein through the endoplasmic reticulum and to the cell surface (12). Additionally, we found that all four target gene plasmids could be mixed and delivered to the same cell without evidence of interference (data not shown), thus alleviating complications in the manufacturing process of the vaccine (i.e., separate gene delivery).…”

Section: Discussionmentioning

confidence: 99%

“…Combinations of the MV and EV immunogens have been shown to elicit more complete protection than that elicited by vaccination with EV or MV targets alone (9,15,16). We have focused on a gene-based molecular vaccine, termed 4pox, targeting the EV immunogens A33 and B5 plus the MV targets L1 and A27 (11,12,(15)(16)(17). This vaccine protects mice and nonhuman primates from lethal vaccinia virus (VACV) or monkeypox virus (MPXV) challenges, respectively (16,17,19).…”

mentioning

confidence: 99%

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Evaluating the Orthopoxvirus Type I Interferon-Binding Molecule as a Vaccine Target in the Vaccinia Virus Intranasal Murine Challenge Model

Golden

Hooper

2010

Clin Vaccine Immunol

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The biological threat imposed by orthopoxviruses warrants the development of safe and effective vaccines. We developed a candidate orthopoxvirus DNA-based vaccine, termed 4pox, which targets four viral structural components, A33, B5, A27, and L1. While this vaccine protects mice and nonhuman primates from lethal infections, we are interested in further enhancing its potency. One approach to enhance potency is to include additional orthopoxvirus immunogens. Here, we investigated whether vaccination with the vaccinia virus (VACV) interferon (IFN)-binding molecule (IBM) could protect BALB/c mice against lethal VACV challenge. We found that vaccination with this molecule failed to significantly protect mice from VACV when delivered alone. IBM modestly augmented protection when delivered together with the 4pox vaccine. All animals receiving the 4pox vaccine plus IBM lived, whereas only 70% of those receiving a single dose of 4pox vaccine survived. Mapping studies using truncated mutants revealed that vaccine-generated antibodies spanned the immunoglobulin superfamily domains 1 and 2 and, to a lesser extent, 3 of the IBM. These antibodies inhibited IBM cell binding and IFN neutralization activity, indicating that they were functionally active. This study shows that DNA vaccination with the VACV IBM results in a robust immune response but that this response does not significantly enhance protection in a high-dose challenge model.

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Improved expression of secretory and trimeric proteins in mammalian cells via the introduction of a new trimer motif and a mutant of the tPA signal sequence

Wang

Song

et al. 2011

Appl Microbiol Biotechnol

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Ideal immunogenicity in antigens is a prerequisite to eliciting a sufficiently strong immune and memory response via either DNA or protein vaccines. To improve immunogenicity, efforts have focused on high-level expression of target proteins and on maintaining their natural conformations. In the present work, two trimer motifs (MTQ and MTI) were designed and introduced into a plasmid vector with the tissue plasminogen activator signal peptide (tPA-SP). Next, we examined the efficacy and the efficiency of the two motifs as well as the introduction of tPA-SP and its mutant forms, 22P/A and 22P/G, in facilitating the secretory expression of trimeric proteins in mammalian cells. We found that both trimer motifs could produce the target protein in a trimeric form at a high level. Introduction of tPA-SP 22P/A markedly increased the secretory expression level. The combination of the trimer motif, MTQ, and the signal peptide, 22P/A, may serve as a universal mammalian vector for producing trimeric proteins in vaccine development.

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Targeting the vaccinia virus L1 protein to the cell surface enhances production of neutralizing antibodies

Cited by 36 publications

References 47 publications

Immunogenicity and Protective Efficiency in Mice of a Smallpox DNA Vaccine Candidate

Immunogenicity and Protective Efficiency in Mice of a Smallpox DNA Vaccine Candidate

Evaluating the Orthopoxvirus Type I Interferon-Binding Molecule as a Vaccine Target in the Vaccinia Virus Intranasal Murine Challenge Model

Improved expression of secretory and trimeric proteins in mammalian cells via the introduction of a new trimer motif and a mutant of the tPA signal sequence

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