Journal: Applied Microbiology and Biotechnology

Article Title: Development and characterization of mouse monoclonal antibodies targeting to distinct epitopes of Zika virus envelope protein for specific detection of Zika virus

doi: 10.1007/s00253-021-11364-1

Figure Lengend Snippet: Alignment of the binding epitopes of mAbs A1 ( a ), B1 ( b ), C1 ( c ), and 9E-1 ( d ) with other flavivirus E proteins. DENV, dengue virus; WNV, West Nile virus; YFV, yellow fever virus; JEV, Japanese encephalitis virus. The NCBI accession numbers of the E proteins derived for the indicated virus strains are provided in the parentheses. e The recombinant E proteins of ZIKV (2 μg), DENV2 (1.75 μg), DENV3 (0.42 μg), and DENV4 (0.78 μg) were separated on SDS-PAGE and then transferred to a PVDF membrane for performing the WB analysis with mAbs Anti-His, A1, B1, C1, or 9E-1, respectively. The numbers labeled on the left of the designated ticks are the molecular weight standards in kDa unit

Article Snippet: The recombinant DENV serotype 2 (DENV2) E protein (GenBank: AAC59274.1), serotype 3 (DENV3) E protein (GenBank: AAA99437.1), and serotype 4 (DENV4) E protein (GenBank: AAX48017.1) were purchased from Sino Biological Inc. (Catalog Number: 40471-V08B, 40532-V08H1, and 40533-V08B2, respectively).

Techniques: Binding Assay, Virus, Derivative Assay, Recombinant, SDS Page, Membrane, Labeling, Molecular Weight

Journal: Emerging Microbes & Infections

Article Title: Convalescent patient-derived monoclonal antibodies targeting different epitopes of E protein confer protection against Zika virus in a neonatal mouse model

doi: 10.1080/22221751.2019.1614885

Figure Lengend Snippet: Binding and neutralizing activities of E-targeted mAbs.

Article Snippet: DENV1 EDIII protein (Cat. no. 40531-V08B), DENV2 EDIII protein (Cat. no. 40471-V08Y3), and DENV4 E protein (Cat. no. 40533-V08B2) were also purchased from Sino Biological Inc.

Techniques: Binding Assay, Neutralization

Journal: Emerging Microbes & Infections

Article Title: Convalescent patient-derived monoclonal antibodies targeting different epitopes of E protein confer protection against Zika virus in a neonatal mouse model

doi: 10.1080/22221751.2019.1614885

Figure Lengend Snippet: Binding and neutralizing activities of E-targeted mAbs.

Article Snippet: DENV1 EDIII protein (Cat. no. 40531-V08B), DENV2 EDIII protein (Cat. no. 40471-V08Y3), and DENV4 E protein (Cat. no. 40533-V08B2) were also purchased from Sino Biological Inc.

Techniques: Binding Assay, Neutralization

Journal: Science signaling

Article Title: Amino acid starvation enhances vaccine efficacy by augmenting neutralizing antibody production

doi: 10.1126/scisignal.aav4717

Figure Lengend Snippet: (A) Experimental outline of HF or vehicle control DMSO treatment and DENV-2 envelope domain III (DENVrEDIII) protein immunization. (B) Proliferation analysis by 3H thymidine incorporation in antigen specific T-cells from splenocytes of mice 28 days after immunization that were restimulated with DENVrEDIII protein. Data are means ± SEM of 12 mice per treatment group from 2 independent experiments.(C to J) Flow cytometry analysis of DENVrEDIII-specific CD8+(C to F) and CD4+(G to J) T-cell responses in blood (D and H), spleen (E and I) and lymph node (F and J) after treatment and immunization, as indicated. FACS plots (C and G) are representative of 2 independent experiments. Quantification of the percentage of IFN-γ producing T-cells (D to F, and H to J) are means ± SEM of 12 mice per treatment group from 2 experiments. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 by two tailed unpaired student t test (B) and Mann-Whitney U test (D, E, F, H, I and J).

Article Snippet: Purification of recombinant DENV envelope domain III protein (DENVrEDIII) The optimized envelope domain III sequence for all the four DENV serotypes (DENV-1, 2, 3 and 4) were cloned into bacterial expression vector pET-28a by Genescript (USA).

Techniques: Control, Flow Cytometry, Two Tailed Test, MANN-WHITNEY

Journal: Science signaling

Article Title: Amino acid starvation enhances vaccine efficacy by augmenting neutralizing antibody production

doi: 10.1126/scisignal.aav4717

Figure Lengend Snippet: (A) Flow cytometry analysis of antigen-specific CD8+IFN-γ+ andCD4+IFN-γ+ T-cell kinetics in PBMCs of mice treated with DMSO or HF and immunized with DENV-2 envelope domain III after 14 and 28 days. Data are means ± SEM of 12 mice/group from 2 independent experiments. (B to D) Flow cytometry analysis of DENVrEDIII-specific polyfunctional CD8+ (B) and CD4+(C) T-cells in blood, spleen and lymph node of immunized mice. The frequency of double cytokine (IFN-γ and IL-2) producing cells with means (bar) ± SEM (B and C) and pie charts of the frequency of all cytokine producing T-cells (D) are from 12 mice/group from 2 independent experiments.*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001by 2-way ANOVA with Bonferroni post-hoc between DENVrEDIII+DMSO and DENVrEDIII+HF immunized groups(A), Mann-Whitney U test(B and C)

Article Snippet: Purification of recombinant DENV envelope domain III protein (DENVrEDIII) The optimized envelope domain III sequence for all the four DENV serotypes (DENV-1, 2, 3 and 4) were cloned into bacterial expression vector pET-28a by Genescript (USA).

Techniques: Flow Cytometry, MANN-WHITNEY

Journal: Science signaling

Article Title: Amino acid starvation enhances vaccine efficacy by augmenting neutralizing antibody production

doi: 10.1126/scisignal.aav4717

Figure Lengend Snippet: ELISA analysis of the amounts of IFN-γ, IL-12p40 and TNF-α produced bysplenocytes and lymph node cells from immunized mice after re-stimulation in vitro with DENVrEDIII for 72hr. Spleen and lymph nodes were collected from mice at 28th day post-secondary immunization. Data are means ± SEM from 12 mice per treatment group from 2 independent experiments. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 by two tailed unpaired student t test.

Article Snippet: Purification of recombinant DENV envelope domain III protein (DENVrEDIII) The optimized envelope domain III sequence for all the four DENV serotypes (DENV-1, 2, 3 and 4) were cloned into bacterial expression vector pET-28a by Genescript (USA).

Techniques: Enzyme-linked Immunosorbent Assay, Produced, In Vitro, Two Tailed Test

Journal: Science signaling

Article Title: Amino acid starvation enhances vaccine efficacy by augmenting neutralizing antibody production

doi: 10.1126/scisignal.aav4717

Figure Lengend Snippet: (A) ELISA analysis of DENVrEDIII specific total IgG, IgG2a, IgG2b, and IgG1 in the serum of mice14 days after primary immunization and 28 days after secondary immunization. Data are means ± SEM of 12 mice/group from 2 independent experiments. (B and C) BIAcore SPR analysis of DENVrEDIII protein binding by pooled serum samples from mice preconditioned with HF or DMSO 28 days after immunization. Sensogram trace of the DENVrEDIII-specific antibody binding affinity (B) and correlation of the Maximal Response Unit (RUmax) with the dissociation constant (C, upper) are representative of 2 independent experiments. Quantified avidity scores(C, lower) are means ± SEM from 2 independent experiments on pooled serum samples from 10 mice/group assayed in triplicate.(D) DENV-2 virus neutralization assay on serum samples collected from mice 28 days after immunization. The 50% focus reduction neutralization titre (FRNT50) data are means ± SEM of 10 mice from 2 independent experiments assayed in triplicate.*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, by one-way ANOVA with Bonferroni post-hoc test (A), two tailed unpaired student t test(C) and Mann-Whitney U test (D).

Article Snippet: Purification of recombinant DENV envelope domain III protein (DENVrEDIII) The optimized envelope domain III sequence for all the four DENV serotypes (DENV-1, 2, 3 and 4) were cloned into bacterial expression vector pET-28a by Genescript (USA).

Techniques: Enzyme-linked Immunosorbent Assay, Protein Binding, Binding Assay, Virus, Neutralization, Two Tailed Test, MANN-WHITNEY

Journal: Science signaling

Article Title: Amino acid starvation enhances vaccine efficacy by augmenting neutralizing antibody production

doi: 10.1126/scisignal.aav4717

Figure Lengend Snippet: (A)ELISA of total IgG specific for all four DENV serotypes in the serum of mice immunized with a tetravalent combination of DENVrEDIII protein 14 days after primary immunization and 28 days after secondary immunization. Data are means ± SEMof10 mice/group from 2 independent experiments. (B)DENV serotype neutralization assay on serum from tDENVrEDIII immunized mice. The 50% focus reduction neutralization titre (FRNT50) data are means ± SEM of 10 mice from 2 independent experiments. *P<0.05, **P<0.01 by one-way ANOVA with Bonferroni post-hoc test (A) and Mann-Whitney U test (B).

Article Snippet: Purification of recombinant DENV envelope domain III protein (DENVrEDIII) The optimized envelope domain III sequence for all the four DENV serotypes (DENV-1, 2, 3 and 4) were cloned into bacterial expression vector pET-28a by Genescript (USA).

Techniques: Enzyme-linked Immunosorbent Assay, Neutralization, MANN-WHITNEY

Journal: Science signaling

Article Title: Amino acid starvation enhances vaccine efficacy by augmenting neutralizing antibody production

doi: 10.1126/scisignal.aav4717

Figure Lengend Snippet: (A and B) Confocal microscopy imaging of the GL-7+(red), B220+(green) IgG+(blue) GC B-cells in lymph node sections from mice treated with HF or DMSO and immunized with DENVrEDIII protein. Images (A) are representative of 2 independent experiments. Quantified data (B) are means ± SEM of 8 mice/condition from all experiments. (C) Flow cytometry analysis of lymph node GC-B cells frequency in DENVrEDIII immunized mice at the indicated time-points. Data are means ± SEM of 8 mice per group at each time-point from 2 independent experiments.(D) ELISA analysis of IL-10 production by splenocytes and lymph node cells from mice 28 days after secondary immunization that were restimulated with DENVrEDIII. Data are means ± SEM of 10 mice/group from 2 independent experiments. (E) qRT-PCR analysis of the indicated gene expression in lymph node cells restimulated with DENVrEDIII for 24hr. Heatmap of statistically significant changes are from the analysis of 10 biological replicates from 2 independent experiments. *P<0.05, **P< 0.01, ***P<0.001, ****P<0.0001by two tailed unpaired student t test (B and D) and 2-way ANOVA with Bonferroni Post-hoc test (C).

Article Snippet: Purification of recombinant DENV envelope domain III protein (DENVrEDIII) The optimized envelope domain III sequence for all the four DENV serotypes (DENV-1, 2, 3 and 4) were cloned into bacterial expression vector pET-28a by Genescript (USA).

Techniques: Confocal Microscopy, Imaging, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Gene Expression, Two Tailed Test

Journal: Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases

Article Title: Aedes aegypti vector competence studies: A review

doi: 10.1016/j.meegid.2018.11.009

Figure Lengend Snippet: Summary of vector competence estimates across Ae. aegypti geographic populations to 1) DENVs, 2) ZIKV, 3) YFV; 4) CHIKV; 5) dual-infections and 6) infections with arboviruses other than DENVs, YFV, ZIKV and CHIKV.

Article Snippet: , Ho Chi Minh City , DENV-2 Bangkok 1974 , BM 3 , 10 8.2 , IR 96,16 +/− 3.35.

Techniques: Plasmid Preparation, Virus, Infection, Mutagenesis, Marker, Standard Deviation, Isolation, Western Blot, Northern Blot