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. 2020 Nov 30;11:6121. doi: 10.1038/s41467-020-19819-1

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Balb/c mice (n = 5) immunized twice in a 3-week interval with 5 × 10⁷ PFU of the single and double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific humoral immune responses. a, b Binding antibodies. S-, RBD-, and N-specific binding antibodies induced by the vaccine vectors were evaluated after the first (a) and second (b) immunization by ELISA. As a comparison, antigen-specific endpoint titers measured in n = 19 plasma samples from SARS-CoV-2 convalescent individuals (Fig. S4) were added in a and b. Data are presented as mean values + SD. One-way ANOVA with Tukey’s multiple comparison test was used to evaluate the differences between binding antibody end-point titers. c IgG2a/IgG1 isotype ratio. S-, RBD-, and N-specific binding antibodies of the IgG2a and IgG1 isotype were measured after the second immunization using 1:10,000 serum dilution, and absorbance reading was used to calculate IgG2a/IgG1 antibody ratio. Lines represent median values. One-way ANOVA with Dunnett’s multiple comparison test was used to compare each group mean IgG2a/IgG1 ratio to a ratio of 1 (balanced Th1/Th2 response). d–g NAb responses. SARS-CoV-2-specific NAb (NT90 titer) induced by the vaccine vectors were measured after the first (d, f) and second (e, g) immunization against SARS-CoV-2 pseudovirus (pv) (d, e) or infectious SARS-CoV-2 virus (f, g) in pooled sera of immunized mice. Shown is the average NT90 measured in duplicate (d, e) or triplicate (f, g) infection. N/A = failed quality control of the samples. Dotted lines indicate lowest antibody dilution included in the analysis. h SARS-CoV-2/SARS-CoV-2pv correlation analysis. Correlation analysis of NT90 measured in mouse sera after one and two immunizations using infectious SARS-CoV-2 virus and SARS-CoV-2pv. Pearson correlation coefficient (r) was calculated in h; *0.05 < p ≪0.01, **0.01 < p < 0.001, ***0.001 < p < 0.0001, ****p < 0.0001; ns = not significant. Source data are provided as a Source Data file.