Journal: JCI insight

Article Title: Antibody effector analysis of prime versus prime-boost immunizations with a recombinant measles-vectored chikungunya virus vaccine.

doi: 10.1172/jci.insight.151095

Figure Lengend Snippet: Figure 1. Study design and characterization of the CHIK-specific IgG response elicited by MV-CHIK prime and prime-boost vaccination strategies. (A) Par- ticipants were vaccinated with either MV-CHIK prime at day 0 (P only) or prime (at month –1) and boost (at day 0) (P+B), and serum antibody responses were compared at 1 and 6 months after vaccination. MV-vaccinated groups served as controls. (B–F) Total IgG binding to CHIKV virus was measured using ELISA and compared (B) between groups and (C) at 1 and 6 months for paired MV-CHIK–vaccinated samples. (D) Fold change in IgG (mean ± 95% CI) in response to CHIKV virus was compared between MV-CHIK P only and P+B. IgG binding to 2 CHIK surface glycoproteins, E1 and E2, was measured using Luminex immu- noassay and compared between groups at (E) 1 month and (F) 6 months after vaccination. (G) Distribution of the CHIK antigens targeted by the antibody response was estimated by the ratio of E2-specific to E1-specific IgG (mean ± 95% CI) and compared between prime and prime-boost MV-CHIK vaccinations. IgG responses between groups were compared using Mann-Whitney U test, while paired analysis between groups at 1 and 6 months was conducted using paired Wilcoxon signed-rank test (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: ELISA plates were then washed and probed with the relevant HRP-conjugated secondary antibody for detection of human IgG (Santa Cruz, sc-2907) and IgG1, IgG2, IgG3, and IgG4 (Southern Biotech, 9054-05, 9060-05, 9210-05, and 9200-05, respectively).

Techniques: Binding Assay, Virus, Enzyme-linked Immunosorbent Assay, Luminex, MANN-WHITNEY

Journal: JCI insight

Article Title: Antibody effector analysis of prime versus prime-boost immunizations with a recombinant measles-vectored chikungunya virus vaccine.

doi: 10.1172/jci.insight.151095

Figure Lengend Snippet: Figure 2. Avidity and neutralizing capacity of CHIK virus–specific antibody responses elicited by MV-CHIK prime and prime-boost immunization. (A) Binding strength of CHIK virus–specific antibodies, as indicated by avidity index estimations following 4, 6, and 8 M urea washes. (B) CHIK virus neutralization (measured by PRNT assay) and (C) neutralization normalized to total IgG (mean ± 95% CI) compared between vaccinated groups. (D and E) Longitudinal assessment of the CHIKV-neutralizing antibody response using (D) paired analysis between 1 and 6 months and (E) fold change in CHIKV neutralization from 1 to 6 months (mean ± 95% CI) in MV-CHIK prime (P) and prime-boost (P+B) immunization. Avidity index and PRNT50 titers were compared between groups using Mann-Whitney U test, and analysis between groups at 1 and 6 months was conducted using paired Wilcoxon signed-rank test (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: ELISA plates were then washed and probed with the relevant HRP-conjugated secondary antibody for detection of human IgG (Santa Cruz, sc-2907) and IgG1, IgG2, IgG3, and IgG4 (Southern Biotech, 9054-05, 9060-05, 9210-05, and 9200-05, respectively).

Techniques: Virus, Binding Assay, Neutralization, Plaque Reduction Neutralization Test, MANN-WHITNEY

Journal: JCI insight

Article Title: Antibody effector analysis of prime versus prime-boost immunizations with a recombinant measles-vectored chikungunya virus vaccine.

doi: 10.1172/jci.insight.151095

Figure Lengend Snippet: Figure 3. IgG subclass distribution of CHIK virus–specific antibody responses following MV-CHIK prime and prime-boost vaccinations. (A) IgG1, IgG2, IgG3, and IgG4 CHIK virus–binding endpoint titers (measured by ELISA) in MV-CHIK–vaccinated individuals. (B and C) Longitudinal analysis of CHIK virus–specific IgG1 and IgG3. (B) Endpoint titers at 1 and 6 months. (C) Fold change in antibody titers from 1 to 6 months (mean ± 95% CI). (D–G) The ratio of each IgG subclass (IgG1, IgG2, IgG3, and IgG4) to total IgG, and (H) the ratio of IgG1 to IgG2 (mean ± 95% CI). IgG subclass titers were compared between groups using Mann-Whitney U test, and comparisons between groups at 1 and 6 months used paired Wilcoxon signed-rank test (**P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: ELISA plates were then washed and probed with the relevant HRP-conjugated secondary antibody for detection of human IgG (Santa Cruz, sc-2907) and IgG1, IgG2, IgG3, and IgG4 (Southern Biotech, 9054-05, 9060-05, 9210-05, and 9200-05, respectively).

Techniques: Virus, Binding Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: JCI insight

Article Title: Antibody effector analysis of prime versus prime-boost immunizations with a recombinant measles-vectored chikungunya virus vaccine.

doi: 10.1172/jci.insight.151095

Figure Lengend Snippet: Figure 4. Antibody-dependent cellular phagocytosis of CHIK virus by a monocytic cell line following vaccination with MV-CHIK. (A) Phagocytic scores (measured using a whole-virus phagocytic assay) compared between vaccinated groups. (B) Paired longitudinal antibody-dependent cellular phagocytosis (ADCP) responses compared at 1 and 6 months. (C) Nor- malized phagocytic scores (i.e., phagocytic scores normalized to CHIKV-specific IgG titers) compared between P- and P+B MC-CHIK–vaccinated groups. Phagocytic scores and normalized phagocytic scores were compared between groups using Mann-Whitney U test, and paired analysis between groups at 1 and 6 months was conducted using paired Wilcoxon signed- rank test (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: ELISA plates were then washed and probed with the relevant HRP-conjugated secondary antibody for detection of human IgG (Santa Cruz, sc-2907) and IgG1, IgG2, IgG3, and IgG4 (Southern Biotech, 9054-05, 9060-05, 9210-05, and 9200-05, respectively).

Techniques: Virus, MANN-WHITNEY

Journal: JCI insight

Article Title: Antibody effector analysis of prime versus prime-boost immunizations with a recombinant measles-vectored chikungunya virus vaccine.

doi: 10.1172/jci.insight.151095

Figure Lengend Snippet: Figure 5. NK-mediated ADCC following MV-CHIK vaccination. ADCC was assessed through the proxy measures of ADNKA degranulation (CD107α) and cytokine release (IFN-γ). (A) Percentage of CD107α+ cells and (B) IFN-γ+ cells in vaccinated groups. Paired longitudinal analysis of the percentage of (C) CD107α+ cells and (D) IFN-γ+ cells at 1 and 6 months after vaccination. Normalized percentage of (E) CD107α+ cells and (F) IFN-γ+ cells (to CHIKV-specific IgG titers) in P- and P+B MV-CHIK–vaccinated groups. Percentages and normalized percentages of CD17α+ and IFN-γ+ were compared between groups using Mann-Whitney U test, and paired analysis between groups at 1 and 6 months was conducted using paired Wilcoxon signed-rank test (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: ELISA plates were then washed and probed with the relevant HRP-conjugated secondary antibody for detection of human IgG (Santa Cruz, sc-2907) and IgG1, IgG2, IgG3, and IgG4 (Southern Biotech, 9054-05, 9060-05, 9210-05, and 9200-05, respectively).

Techniques: MANN-WHITNEY

Journal: JCI insight

Article Title: Antibody effector analysis of prime versus prime-boost immunizations with a recombinant measles-vectored chikungunya virus vaccine.

doi: 10.1172/jci.insight.151095

Figure Lengend Snippet: Figure 6. Multidimensional unsupervised analysis of MV-CHIK–vaccinated groups and measured antibody phenotypes. (A and B) Principal component analysis (PCA) was conducted on MV-CHIK vaccine–induced antibody responses following prime (P) and prime-boost (P+B) regiments at 1 and 6 months. (A) Individual plot with vaccine groups highlighted in different colors. Ellipses in the PCA individual plot represent the confidence ellipses around the centroid (marked by the larger symbol for each group) of each vaccine group. (B) Circle plot highlighting the variables contributing to the spread of points. (C) Cor- relation matrix with hierarchical clustering of variables utilized in the PCA analysis. Variables included in the PCA and hierarchical correlation matrix include neutralization (PRNT50 titers); virus-binding total IgG, IgG1, IgG2, IgG3, and IgG4 (endpoint titers); E1-binding IgG (MFI); E2-binding IgG (MFI); avidity at 8 M (AI); ADCP (phagocytic score); and ADCC (ADNKA degranulation as percentage of CD107α+ cells, and ADNKA cytokine release as percentage of IFN-γ+ cells).

Article Snippet: ELISA plates were then washed and probed with the relevant HRP-conjugated secondary antibody for detection of human IgG (Santa Cruz, sc-2907) and IgG1, IgG2, IgG3, and IgG4 (Southern Biotech, 9054-05, 9060-05, 9210-05, and 9200-05, respectively).

Techniques: Neutralization, Virus, Binding Assay