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e2 chikv ectodomain consensus protein  (GenScript corporation)

 
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    GenScript corporation e2 chikv ectodomain consensus protein
    Homologous DNA or <t>E2*</t> <t>CHIKV</t> protein prime-boost immunization induces robust humoral responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with scDEC-E2 (a DC-targeted DNA vaccine) followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). The control group received empty pVAX vector and poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. ( b ) E2*-Specific IgG subclasses after boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed by one-way ANOVA followed by Tukey post-hoc test. Data represent the mean ± SD and are representative from 3 independent experiments. (a,b) statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    E2 Chikv Ectodomain Consensus Protein, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e2 chikv ectodomain consensus protein/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    e2 chikv ectodomain consensus protein - by Bioz Stars, 2026-04
    90/100 stars

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    1) Product Images from "Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity"

    Article Title: Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms241310517

    Homologous DNA or E2* CHIKV protein prime-boost immunization induces robust humoral responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with scDEC-E2 (a DC-targeted DNA vaccine) followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). The control group received empty pVAX vector and poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. ( b ) E2*-Specific IgG subclasses after boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed by one-way ANOVA followed by Tukey post-hoc test. Data represent the mean ± SD and are representative from 3 independent experiments. (a,b) statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    Figure Legend Snippet: Homologous DNA or E2* CHIKV protein prime-boost immunization induces robust humoral responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with scDEC-E2 (a DC-targeted DNA vaccine) followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). The control group received empty pVAX vector and poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. ( b ) E2*-Specific IgG subclasses after boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed by one-way ANOVA followed by Tukey post-hoc test. Data represent the mean ± SD and are representative from 3 independent experiments. (a,b) statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Techniques Used: In Vivo, Electroporation, Recombinant, Control, Plasmid Preparation, Incubation

    Immunization with vaccines encoding E2 CHIKV induces robust humoral immune responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with 100 μg of a DC-targeted scDEC-E2 DNA vaccine followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). For the heterologous DNA prime-protein boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by a boost with E2* recombinant protein + poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. a- statistical analysis in comparison to the first dose. ( b ) E2*-specific IgG subclasses after the boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD and are representative of 2 independent experiments. ( a ) Statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    Figure Legend Snippet: Immunization with vaccines encoding E2 CHIKV induces robust humoral immune responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with 100 μg of a DC-targeted scDEC-E2 DNA vaccine followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). For the heterologous DNA prime-protein boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by a boost with E2* recombinant protein + poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. a- statistical analysis in comparison to the first dose. ( b ) E2*-specific IgG subclasses after the boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD and are representative of 2 independent experiments. ( a ) Statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Techniques Used: Vaccines, In Vivo, Electroporation, Recombinant, Comparison, Incubation

    Immunization with vaccines encoding E2 CHIKV elicits robust T and B cell responses. C57BL/6 mice were immunized as described in (immunization strategy displayed in ). ( a ) Fifteen days after the boost, spleen cells were cultured in the presence of individual peptides from the E2* recombinant protein (10 μg/mL) to evaluate the number of IFN-γ-producing cells using the ELISpot assay. SFU: spot forming units. a, b, c, d represent statistical significance between the homologous and respective heterologous prime-boost strategies. #, €, &, $ represent the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous immunization strategy. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) by ELISpot. Statistical analysis was performed with the two-way ANOVA followed by Bonferroni’s post-hoc ( a ), or with the one-way ANOVA followed by the Tukey post-hoc test ( b ). Data represent the mean ± SD and are representative of 2 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    Figure Legend Snippet: Immunization with vaccines encoding E2 CHIKV elicits robust T and B cell responses. C57BL/6 mice were immunized as described in (immunization strategy displayed in ). ( a ) Fifteen days after the boost, spleen cells were cultured in the presence of individual peptides from the E2* recombinant protein (10 μg/mL) to evaluate the number of IFN-γ-producing cells using the ELISpot assay. SFU: spot forming units. a, b, c, d represent statistical significance between the homologous and respective heterologous prime-boost strategies. #, €, &, $ represent the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous immunization strategy. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) by ELISpot. Statistical analysis was performed with the two-way ANOVA followed by Bonferroni’s post-hoc ( a ), or with the one-way ANOVA followed by the Tukey post-hoc test ( b ). Data represent the mean ± SD and are representative of 2 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Techniques Used: Vaccines, Cell Culture, Recombinant, Enzyme-linked Immunospot

    Immunization in the presence of AS03 adjuvant elicits robust humoral responses with a strong neutralizing ability. C57BL/6 mice were immunized intramuscularly twice 15 days apart with 100 μg of the non-targeted pVAX-E2 DNA vaccine or the DC-targeted scDEC-E2 DNA vaccine followed by electroporation, or with 10 μg of E2* recombinant protein + AS03 subcutaneously (immunization strategy displayed in ). For the heterologous prime-boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by E2* recombinant protein + AS03. The control group received the empty pVAX vector and AS03. Blood samples were collected 14 days after each immunization to evaluate the antibody response. ( a ) Vero E6 cells were infected with CHIKV virus (MOI = 0.1) for 20 h, incubated with pooled sera, followed by donkey-anti mouse IgG-Alexa Fluor 488 and DAPI staining. ( b ) Total E2*-specific IgG titers. ( c ) E2*-specific IgG subclasses after the boost on a logarithm scale. ( d ) Antibody affinity from pooled sera after incubation with increasing concentrations of ammonium thiocyanate. ( e ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. a- statistical significance conducted when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    Figure Legend Snippet: Immunization in the presence of AS03 adjuvant elicits robust humoral responses with a strong neutralizing ability. C57BL/6 mice were immunized intramuscularly twice 15 days apart with 100 μg of the non-targeted pVAX-E2 DNA vaccine or the DC-targeted scDEC-E2 DNA vaccine followed by electroporation, or with 10 μg of E2* recombinant protein + AS03 subcutaneously (immunization strategy displayed in ). For the heterologous prime-boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by E2* recombinant protein + AS03. The control group received the empty pVAX vector and AS03. Blood samples were collected 14 days after each immunization to evaluate the antibody response. ( a ) Vero E6 cells were infected with CHIKV virus (MOI = 0.1) for 20 h, incubated with pooled sera, followed by donkey-anti mouse IgG-Alexa Fluor 488 and DAPI staining. ( b ) Total E2*-specific IgG titers. ( c ) E2*-specific IgG subclasses after the boost on a logarithm scale. ( d ) Antibody affinity from pooled sera after incubation with increasing concentrations of ammonium thiocyanate. ( e ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. a- statistical significance conducted when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Techniques Used: Adjuvant, Electroporation, Recombinant, Control, Plasmid Preparation, Infection, Virus, Incubation, Staining

    Vaccines containing E2 CHIKV elicit cellular immune responses with a cytotoxic profile. C57BL/6 mice (n = 6) were immunized as described in (immunization strategy displayed in ). Fifteen days after the boost, mice were euthanized and spleen and draining lymph nodes were removed. ( a ) Specific IFN-γ production was examined with ELISpot against individual peptides. (a, b)) represents statistical significance between the homologous and heterologous strategies with the same vaccine used as a prime. #, €, & indicate the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous regimen. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) with ELISpot ( c ) In vivo cytotoxicity assay against target cells pulsed with the E2 355–364 peptide. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    Figure Legend Snippet: Vaccines containing E2 CHIKV elicit cellular immune responses with a cytotoxic profile. C57BL/6 mice (n = 6) were immunized as described in (immunization strategy displayed in ). Fifteen days after the boost, mice were euthanized and spleen and draining lymph nodes were removed. ( a ) Specific IFN-γ production was examined with ELISpot against individual peptides. (a, b)) represents statistical significance between the homologous and heterologous strategies with the same vaccine used as a prime. #, €, & indicate the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous regimen. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) with ELISpot ( c ) In vivo cytotoxicity assay against target cells pulsed with the E2 355–364 peptide. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Techniques Used: Vaccines, Enzyme-linked Immunospot, Cell Culture, In Vivo, Cytotoxicity Assay



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    e2 chikv ectodomain consensus protein  (GenScript corporation)
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    GenScript corporation e2 chikv ectodomain consensus protein
    Homologous DNA or <t>E2*</t> <t>CHIKV</t> protein prime-boost immunization induces robust humoral responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with scDEC-E2 (a DC-targeted DNA vaccine) followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). The control group received empty pVAX vector and poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. ( b ) E2*-Specific IgG subclasses after boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed by one-way ANOVA followed by Tukey post-hoc test. Data represent the mean ± SD and are representative from 3 independent experiments. (a,b) statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    E2 Chikv Ectodomain Consensus Protein, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e2 chikv ectodomain consensus protein/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    e2 chikv ectodomain consensus protein - by Bioz Stars, 2026-04
    90/100 stars
    		  Buy from Supplier

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    Homologous DNA or E2* CHIKV protein prime-boost immunization induces robust humoral responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with scDEC-E2 (a DC-targeted DNA vaccine) followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). The control group received empty pVAX vector and poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. ( b ) E2*-Specific IgG subclasses after boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed by one-way ANOVA followed by Tukey post-hoc test. Data represent the mean ± SD and are representative from 3 independent experiments. (a,b) statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity

    doi: 10.3390/ijms241310517

    Figure Lengend Snippet: Homologous DNA or E2* CHIKV protein prime-boost immunization induces robust humoral responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with scDEC-E2 (a DC-targeted DNA vaccine) followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). The control group received empty pVAX vector and poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. ( b ) E2*-Specific IgG subclasses after boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed by one-way ANOVA followed by Tukey post-hoc test. Data represent the mean ± SD and are representative from 3 independent experiments. (a,b) statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The E2 CHIKV ectodomain consensus protein sequence (aa 1 to 364) lacking the transmembrane region (aa 365 to 422) was generated after the alignment (ClustalW, MegAlign Pro V.17.4.2, DNAStar) of 79 sequences of the Brazilian virus isolates (Genbank Accession Numbers— ) and synthesized by GenScript (Piscataway, NJ, USA).

    Techniques: In Vivo, Electroporation, Recombinant, Control, Plasmid Preparation, Incubation

    Immunization with vaccines encoding E2 CHIKV induces robust humoral immune responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with 100 μg of a DC-targeted scDEC-E2 DNA vaccine followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). For the heterologous DNA prime-protein boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by a boost with E2* recombinant protein + poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. a- statistical analysis in comparison to the first dose. ( b ) E2*-specific IgG subclasses after the boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD and are representative of 2 independent experiments. ( a ) Statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity

    doi: 10.3390/ijms241310517

    Figure Lengend Snippet: Immunization with vaccines encoding E2 CHIKV induces robust humoral immune responses. C57BL/6 mice were either immunized intramuscularly twice with 100 μg of pVAX-E2 (non-targeted DNA vaccine), with 100 μg of a DC-targeted scDEC-E2 DNA vaccine followed by in vivo electroporation, or with 10 μg of E2* recombinant protein + poly (I:C) subcutaneously (immunization strategy displayed in ). For the heterologous DNA prime-protein boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by a boost with E2* recombinant protein + poly (I:C). Blood samples were collected 14 days after each immunization to evaluate the humoral immune response. ( a ) Total E2*-specific IgG titers. a- statistical analysis in comparison to the first dose. ( b ) E2*-specific IgG subclasses after the boost. ( c ) Antibody affinity of pooled mouse sera after incubation with increasing concentrations of ammonium thiocyanate. ( d ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD and are representative of 2 independent experiments. ( a ) Statistical significance when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The E2 CHIKV ectodomain consensus protein sequence (aa 1 to 364) lacking the transmembrane region (aa 365 to 422) was generated after the alignment (ClustalW, MegAlign Pro V.17.4.2, DNAStar) of 79 sequences of the Brazilian virus isolates (Genbank Accession Numbers— ) and synthesized by GenScript (Piscataway, NJ, USA).

    Techniques: Vaccines, In Vivo, Electroporation, Recombinant, Comparison, Incubation

    Immunization with vaccines encoding E2 CHIKV elicits robust T and B cell responses. C57BL/6 mice were immunized as described in (immunization strategy displayed in ). ( a ) Fifteen days after the boost, spleen cells were cultured in the presence of individual peptides from the E2* recombinant protein (10 μg/mL) to evaluate the number of IFN-γ-producing cells using the ELISpot assay. SFU: spot forming units. a, b, c, d represent statistical significance between the homologous and respective heterologous prime-boost strategies. #, €, &, $ represent the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous immunization strategy. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) by ELISpot. Statistical analysis was performed with the two-way ANOVA followed by Bonferroni’s post-hoc ( a ), or with the one-way ANOVA followed by the Tukey post-hoc test ( b ). Data represent the mean ± SD and are representative of 2 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity

    doi: 10.3390/ijms241310517

    Figure Lengend Snippet: Immunization with vaccines encoding E2 CHIKV elicits robust T and B cell responses. C57BL/6 mice were immunized as described in (immunization strategy displayed in ). ( a ) Fifteen days after the boost, spleen cells were cultured in the presence of individual peptides from the E2* recombinant protein (10 μg/mL) to evaluate the number of IFN-γ-producing cells using the ELISpot assay. SFU: spot forming units. a, b, c, d represent statistical significance between the homologous and respective heterologous prime-boost strategies. #, €, &, $ represent the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous immunization strategy. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) by ELISpot. Statistical analysis was performed with the two-way ANOVA followed by Bonferroni’s post-hoc ( a ), or with the one-way ANOVA followed by the Tukey post-hoc test ( b ). Data represent the mean ± SD and are representative of 2 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The E2 CHIKV ectodomain consensus protein sequence (aa 1 to 364) lacking the transmembrane region (aa 365 to 422) was generated after the alignment (ClustalW, MegAlign Pro V.17.4.2, DNAStar) of 79 sequences of the Brazilian virus isolates (Genbank Accession Numbers— ) and synthesized by GenScript (Piscataway, NJ, USA).

    Techniques: Vaccines, Cell Culture, Recombinant, Enzyme-linked Immunospot

    Immunization in the presence of AS03 adjuvant elicits robust humoral responses with a strong neutralizing ability. C57BL/6 mice were immunized intramuscularly twice 15 days apart with 100 μg of the non-targeted pVAX-E2 DNA vaccine or the DC-targeted scDEC-E2 DNA vaccine followed by electroporation, or with 10 μg of E2* recombinant protein + AS03 subcutaneously (immunization strategy displayed in ). For the heterologous prime-boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by E2* recombinant protein + AS03. The control group received the empty pVAX vector and AS03. Blood samples were collected 14 days after each immunization to evaluate the antibody response. ( a ) Vero E6 cells were infected with CHIKV virus (MOI = 0.1) for 20 h, incubated with pooled sera, followed by donkey-anti mouse IgG-Alexa Fluor 488 and DAPI staining. ( b ) Total E2*-specific IgG titers. ( c ) E2*-specific IgG subclasses after the boost on a logarithm scale. ( d ) Antibody affinity from pooled sera after incubation with increasing concentrations of ammonium thiocyanate. ( e ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. a- statistical significance conducted when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity

    doi: 10.3390/ijms241310517

    Figure Lengend Snippet: Immunization in the presence of AS03 adjuvant elicits robust humoral responses with a strong neutralizing ability. C57BL/6 mice were immunized intramuscularly twice 15 days apart with 100 μg of the non-targeted pVAX-E2 DNA vaccine or the DC-targeted scDEC-E2 DNA vaccine followed by electroporation, or with 10 μg of E2* recombinant protein + AS03 subcutaneously (immunization strategy displayed in ). For the heterologous prime-boost, mice received one dose of a DNA vaccine (pVAX-E2 or scDEC-E2) followed by E2* recombinant protein + AS03. The control group received the empty pVAX vector and AS03. Blood samples were collected 14 days after each immunization to evaluate the antibody response. ( a ) Vero E6 cells were infected with CHIKV virus (MOI = 0.1) for 20 h, incubated with pooled sera, followed by donkey-anti mouse IgG-Alexa Fluor 488 and DAPI staining. ( b ) Total E2*-specific IgG titers. ( c ) E2*-specific IgG subclasses after the boost on a logarithm scale. ( d ) Antibody affinity from pooled sera after incubation with increasing concentrations of ammonium thiocyanate. ( e ) For PRNT, pooled sera were incubated with 100 PFU of CHIKV and the NT 50 is displayed. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. a- statistical significance conducted when compared to the first dose. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The E2 CHIKV ectodomain consensus protein sequence (aa 1 to 364) lacking the transmembrane region (aa 365 to 422) was generated after the alignment (ClustalW, MegAlign Pro V.17.4.2, DNAStar) of 79 sequences of the Brazilian virus isolates (Genbank Accession Numbers— ) and synthesized by GenScript (Piscataway, NJ, USA).

    Techniques: Adjuvant, Electroporation, Recombinant, Control, Plasmid Preparation, Infection, Virus, Incubation, Staining

    Vaccines containing E2 CHIKV elicit cellular immune responses with a cytotoxic profile. C57BL/6 mice (n = 6) were immunized as described in (immunization strategy displayed in ). Fifteen days after the boost, mice were euthanized and spleen and draining lymph nodes were removed. ( a ) Specific IFN-γ production was examined with ELISpot against individual peptides. (a, b)) represents statistical significance between the homologous and heterologous strategies with the same vaccine used as a prime. #, €, & indicate the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous regimen. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) with ELISpot ( c ) In vivo cytotoxicity assay against target cells pulsed with the E2 355–364 peptide. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity

    doi: 10.3390/ijms241310517

    Figure Lengend Snippet: Vaccines containing E2 CHIKV elicit cellular immune responses with a cytotoxic profile. C57BL/6 mice (n = 6) were immunized as described in (immunization strategy displayed in ). Fifteen days after the boost, mice were euthanized and spleen and draining lymph nodes were removed. ( a ) Specific IFN-γ production was examined with ELISpot against individual peptides. (a, b)) represents statistical significance between the homologous and heterologous strategies with the same vaccine used as a prime. #, €, & indicate the statistical significance between the pVAX-E2 and scDEC-E2 groups in the homologous regimen. ( b ) Draining lymph node cells were cultured in the presence of E2* to evaluate the number of specific antibody-secreting cells (ASCs) with ELISpot ( c ) In vivo cytotoxicity assay against target cells pulsed with the E2 355–364 peptide. Statistical analysis was performed with the one-way ANOVA followed by the Tukey post-hoc test. Data represent the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The E2 CHIKV ectodomain consensus protein sequence (aa 1 to 364) lacking the transmembrane region (aa 365 to 422) was generated after the alignment (ClustalW, MegAlign Pro V.17.4.2, DNAStar) of 79 sequences of the Brazilian virus isolates (Genbank Accession Numbers— ) and synthesized by GenScript (Piscataway, NJ, USA).

    Techniques: Vaccines, Enzyme-linked Immunospot, Cell Culture, In Vivo, Cytotoxicity Assay