Binding activity and immunogenic characterization of recombinant C-terminal quarter and half of the heavy chain of botulinum neurotoxin serotype A

Yu, Yun-Zhou; Ma, Yao; Chen, Yanxia; Gong, Zheng-Wei; Wang, Shuang; Yu, Wei-Yuan

doi:10.4161/hv.7.10.16763

Cited by 12 publications

(6 citation statements)

References 34 publications

(47 reference statements)

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“…The efficacy of this type of DNA vaccines against BoNT/A also needs to compare with that of the subunit AHc or toxoid vaccines for future clinical trial or clinical use. 37,38 To our knowledge, this is the first report to date demonstrating the potency of IL-4 as an adjuvant on replicon vaccines in both strain mice. Thus, we described a strategy to design and develop efficient vaccines against BoNT/A or other pathogens using one replicon vector to simultaneously co-express antigen and molecular adjuvant.…”

Section: Discussionmentioning

confidence: 80%

“…Pooled sera from each group of mice above were diluted initially 1:8 and then 2-fold for serum neutralization titers as described previously. 37,38 Briefly, mixtures of serial dilutions of sera with 100 LD 50 of BoNT/A (L+1/100) between group differences. Fisher's exact test was used to determine statistical differences in survival between the treatment groups.…”

Section: Methodsmentioning

confidence: 99%

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Enhanced potency of replicon vaccine using one vector to simultaneously co-express antigen and interleukin-4 molecular adjuvant

Wei

et al. 2013

Human Vaccines & Immunotherapeutics

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We evaluated the utility of interleukin-4 (IL-4) as molecular adjuvant of replicon vaccines for botulinum neurotoxin serotype A (BoNT/A) in mouse model. In both Balb/c and C57/BL6 mice that received the plasmid DNA replicon vaccines derived from Semliki Forest virus (SFV) encoding the Hc gene of BoNT/A (AHc), the immunogenicity was significantly modulated and enhanced by co-delivery or co-express of the IL-4 molecular adjuvant. The enhanced potencies were also produced by co-delivery or co-expression of the IL-4 molecular adjuvant in mice immunized with the recombinant SFV replicon particles (VRP) vaccines. In particular, when AHc and IL-4 were co-expressed within the same replicon vaccine vector using dual-expression or bicistronic IRES, the anti-AHc antibody titers, serum neutralization titers and survival rates of immunized mice after challenged with BoNT/A were significantly increased. These results indicate IL-4 is an effective Th2-type adjuvant for the replicon vaccines in both strain mice, and the co-expression replicon vaccines described here may be an excellent candidate for further vaccine development in other animals or humans. Thus, we described a strategy to design and develop efficient vaccines against BoNT/A or other pathogens using one replicon vector to simultaneously co-express antigen and molecular adjuvant.

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Section: Discussionmentioning

confidence: 80%

Section: Methodsmentioning

confidence: 99%

Enhanced potency of replicon vaccine using one vector to simultaneously co-express antigen and interleukin-4 molecular adjuvant

Wei

et al. 2013

Human Vaccines & Immunotherapeutics

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“…It has been demonstrated that genetic fusion of the chemokines MCP-3 and IP-10 to a model self-tumor antigen converts non-immunogenic sFv into a potent immunogen and induces anti-idiotypic antibody responses and protective antitumor immunity [17,22,23]. DNA vaccines encoding gp120 fused with proinflammatory chemoattractants of immature dendritic cells, such as β-defensin 2, monocyte chemoattractant protein-3 (MCP-3) or macrophage-derived chemokine (MDC/ CCL22), elicited anti-gp120 antibodies with high titers of virus-neutralizing activity [19]. A prototype MDC/CCL22-based DNA epitope vaccine expressing a fusion protein was shown to promote a strong anti-inflammatory Th2-polarized humoral immune response in a 3×Tg-AD mouse model [24].…”

Section: Discussionmentioning

confidence: 99%

“…Anti-AHc-C antibodies in the sera from mice in the different groups were detected by ELISA as described previously [19,20]. Briefly, ELISA plates (Corning) were coated overnight at 4°C with 100 μl recombinant AHc-C (2 μg/ml).…”

Section: Detection Of Anti-ahc-c Antibodies In Serummentioning

confidence: 99%

Co-administration of antigen with chemokine MCP-3 or MDC/CCL22 enhances DNA vaccine potency

Xie

Wang²,

Yang³

et al. 2015

Invest New Drugs

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We evaluated the utility of chemokine MCP-3 and MDC/CCL22 as molecular adjuvants of DNA vaccines for botulinum neurotoxin serotype A (BoNT/A) in a Balb/c mouse model. Notably, the immunogenicity of the DNA vaccine against BoNT/A was not enhanced using a fusion of the AHc-C antigen with the MCP-3 or MDC/CCL22. Nevertheless, the potency of the DNA vaccine was significantly modulated and enhanced by co-administration of the AHc-C antigen with MCP-3 or MDC/CCL22. This strategy elicited high levels of humoral immune responses and protection against BoNT/A. The enhanced potency was further boosted by co-administration of the AHc-C antigen with both MCP-3 and MDC/CCL22 in Balb/c mice, but not by co-administration of AHc-C antigen with the MCP-3-MDC/CCL22 fusion. Co-immunization with both the MCP-3 and MDC/CCL22 constructs induced the highest levels of humoral immunity and protective potency against BoNT/A. Our results indicated that MCP-3 and MDC/CCL22 are effective molecular adjuvants of the immune responses induced by the AHc-C-expressing DNA vaccine when delivered by co-administration of the individual chemokines, but not when delivered in the form of a chemokine/antigen fusion. Thus, we describe an alternative strategy to the design and optimization of DNA vaccine constructs based on co-administration of the antigen with the chemokine rather than in the form of a chemokine/antigen fusion.

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“…35 Briefly, ELISA plates were coated with 100 ml of 40 mg/ ml bovine ganglioside GT1b (Sigma) per well and blocked with PBS containing 1% Casein (Sigma) at 25 C for 3 h. After incubation, 100 ml of purified BHc (50 mg /mL) or BSA (50 mg/ mL) was added to different wells for 3 h at 25 C. Unbound BHc protein was removed by three washes each with PBS containing 0.05% Tween-20 (PBST). Plates were incubated for 8 h at 4 C with 100 ml of 1:2,000 dilutions of polyclonal mouse anti-BHc antibodies, washed, and incubated for 0.5 h with goat antimouse IgG-HRP.…”

Section: Ganglioside Bindingmentioning

confidence: 99%

Production and evaluation of a recombinant subunit vaccine against botulinum neurotoxin serotype B using a 293E expression system

Shi²,

Liu³

et al. 2014

Human Vaccines & Immunotherapeutics

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Keywords: Botulinum neurotoxin serotype B, Hc, expression, vaccine, binding activityAlthough Escherichia coli and yeast were commonly used to express recombinant Hc of botulinum neurotoxins, as an alternative, in current study, a 293E expression system was used to express the Hc of botulinum neurotoxin serotype B (BHc) as soluble recombinant protein for experimental vaccine evaluation. Our results demonstrated that the 293E expression system could produce high level of recombinant secreted BHc protein, which was immunorecognized specifically by anti-botulinum neurotoxin serotype B (BoNT/B) sera and showed ganglioside binding activities. The serological response and efficacy of recombinant BHc formulated with aluminum hydroxide adjuvant were evaluated in mice. Immunization with Alhydrogel-formulated BHc subunit vaccine afforded the effective protection against BoNT/B challenge. A frequency-and dose-dependent effect to immunization with BHc subunit vaccine was observed and the ELISA antibody titers correlated well with neutralizing antibody titers and protection. And a solid-phase assay showed that the neutralizing antibodies from the BHc-immunized mice inhibited the binding of BHc to the ganglioside GT1b. Our results also show that the plasmid pABE293SBHc derived of the 293E expression system as DNA vaccine is capable of inducing stronger humoral response and protective efficacy against BoNT/B than the pVAX1SBHc. In summary, immunization with the 293E-expressed BHc protein generates effective immune protection against BoNT/B as E. coli or yeast-expressed BHc, so the efficient expression of botulinum Hc protein for experimental vaccine can be prepared using the 293E expression system.

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Binding activity and immunogenic characterization of recombinant C-terminal quarter and half of the heavy chain of botulinum neurotoxin serotype A

Abstract: (2011) Binding activity and immunogenic characterization of recombinant C-terminal quarter and half of the heavy chain of botulinum neurotoxin serotype A, Human Vaccines, 7

Cited by 12 publications

References 34 publications

Enhanced potency of replicon vaccine using one vector to simultaneously co-express antigen and interleukin-4 molecular adjuvant

Enhanced potency of replicon vaccine using one vector to simultaneously co-express antigen and interleukin-4 molecular adjuvant

Co-administration of antigen with chemokine MCP-3 or MDC/CCL22 enhances DNA vaccine potency

Production and evaluation of a recombinant subunit vaccine against botulinum neurotoxin serotype B using a 293E expression system

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