Non-neutralizing Antibody Responses against VP1 in Enterovirus A, B, C and Rhinovirus A species among Infants and Children in Shanghai

Figures

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  Figure 1. Expression and purification of recombinant EV71, CA16, CB3, PV1 and RV13 VP1 proteins. The relative molecular weights (MW) of the fusion proteins were 35,636 for pET21b-EV71 VP1, 34,242 for pET21b-CA16 VP1, 34,797 for pET21b-CB3 VP1, 34,902 for pET21b-PV1 VP1, 36,192 for pET21b-RV13 VP1. The expressed proteins were purified using Ni-NTA column affinity chromatography.

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  Figure 2. Characteristics of the antibody reactivity against VP1 of EV71, CA16, CB3, PV1 and RV13 in the serum samples of infants and children from eight age groups (1–28-day, 1–6-month, 7–12-month, 13–24-month, 25–36-month, 37–48-month, 49–60-month and 61–72-month age groups). (A) Comparison of the anti-EV71, anti-CA16, anti-CB3, anti-PV1 or anti-RV13 VP1 reactivity in eight age groups. Each symbol represents an individual sample, and the line represents the median of the samples. Statistical significance was tested using the Nemenyi non-parametric test. * indicates p < 0.05, ** indicates p < 0.001. (B) The age-related dynamic change of antibody reactivity against VP1 of EV71, CA16, CB3, PV1 and RV13 in eight age groups.

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  Figure 3. Characteristics of the antibody reactivity against VP1 of EV71, CA16, CB3, PV1 and RV13 in the serum samples from three age groups (1–28-day, 1–12-month and 13–72-month age groups). (A) Comparison of the antibody reactivity against VP1 of EV71, CA16, CB3, PV1 and RV13 among three age groups. Each symbol represents an individual sample. The line represents the median of the samples. (B) Comparison of the median of anti-VP1 reactivity of the five assessed EVs among three age groups. Statistical significance was tested using the Nemenyi non-parametric test or one-way ANOVA analysis. * indicates p < 0.05, ** indicates p < 0.001, “NS” represents no significant difference between the two groups (p > 0.05). In the 13–72-month age group, # represents p < 0.05 between the antibody reactivity against any two VP1 from the five assessed EV VP1 proteins except two pairs: EV71 vs CA16 and CB3 vs PV1.

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  Figure 4. Comparison of the correlation analyses between five assessed EV VP1 proteins based on antibody responses or inhibition to anti-VP1 reactions by various EV VP1 proteins in three age groups (1–28-day, 1–12-month and 13–72-month age groups). (A) Correlation analysis of antibody reactivity against VP1 of EV71, CA16, CB3, PV1 and RV13 in three age groups. (B) Correlation analysis of inhibition to the anti-VP1 reactions of five assessed EVs, EV71, CA16, CB3, PV1 or RV13 by various VP1 proteins in three age groups.

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  Figure 5. Comparison of inhibition activities to anti-VP1 reactions of EV71 (A), CA16 (B), CB3 (C), PV1 (D), and RV13 (E) by five EV VP1 proteins in the 1–28-day, 1–12-month and 13–72-month age groups. The coated antigen was EV71 VP1 (A), CA16 VP1 (B), CB3 VP1 (C), PV1 VP1 (D) and RV13 VP1 (E) respectively. The percent inhibition from the competitive ELISA was plotted on the y-axis with the five inhibitor proteins on the x-axis: VP1 of EV71, CA16, CB3, PV1 and RV13. The line represents the median of the samples. Statistical significance was tested using the Nemenyi non-parametric test. * represents p < 0.05, ** represents p < 0.001.

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  Figure 6. Comparison of inhibitory potencies to anti-VP1 reactions of EV71 (A), CA16 (B), CB3 (C), PV1 (D), and RV13 (E) in three age groups by five EV VP1 proteins, EV71, CA16, CB3, PV1 and RV13 VP1. The coated antigen was EV71 VP1 (A), CA16 VP1 (B), CB3 VP1 (C), PV1 VP1 (D) and RV13 VP1 (E), respectively. The inhibitory protein had been marked above the figure. The percent inhibition from the competitive ELISA was plotted on the y-axis with three age groups on the x-axis. The line represents the median of the samples. Statistical significance was tested using the Nemenyi non-parametric test. * represents p < 0.05, ** represents p < 0.001.