Adenovirus‐Specific T Cells in Adults Are Frequent, Cross‐Reactive to Common Childhood Adenovirus Infections and Boosted by Adenovirus‐Vectored Vaccines

Figures

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  Figure 1. Plot showing serum antibody responses to adenovirus C5 in healthy blood donors. Optical density measurements were made using an ELISA specific for HAdV C5 and were background‐corrected.

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  Figure 2. Analysis of AdV specific IFNγ and IL2 FluoroSpot responses to AdV peptide pools (A) IFNγ FluoroSpot responses to AdV peptide pools covering: an experimentally‐validated set of 50 AdV C2 and 5 peptides (AdSelect), peptide pools covering the entire hexons of human adenoviruses A12, B3, C5, D26, F41 and the hexon of chimpanzee adenovirus Y25 (vector backbone of SARS‐CoV‐2 vaccine ChAdOx1), as well as a polyclonal anti‐CD3/CD28 antibody T cell stimulation as a positive control of PBMC from healthy blood donors, calculated as spot‐forming units (SFU) per 106 PBMC (background corrected). (B) IL2 FluoroSpot responses to AdV peptide pools covering: an experimentally‐validated set of 50 AdV C2 and 5 peptides (AdSelect), pools covering the entire hexons of human adenoviruses A12, B3, C5, D26, F41, the hexon of chimpanzee adenovirus Y25 (vector backbone of SARS‐CoV‐2 vaccine ChAdOx1), as well as a polyclonal anti‐CD3/CD28 antibody T cell stimulation as a positive control of PBMC from healthy blood donors, calculated as spot‐forming units (SFU) per 106 PBMC (background corrected). Significance determined by two‐way ANOVA, corrected for multiple testing. Key: *p < 0.05. The dotted lines…

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  Figure 3. Analysis of AdV specific IFNγ and IL2 FluoroSpot responses to AdV peptide pools (A) IFNγ FluoroSpot responses to AdV peptide pools covering: the pentons of human adenoviruses C5 and D26, and the penton of chimpanzee adenovirus Y25 (vector backbone ChAdOx1) as well as a polyclonal anti‐CD3/CD28 antibody T cell stimulation as a positive control of PBMC from healthy blood donors, calculated as spot‐forming units (SFU) per 106 PBMC (background corrected). (B) IL2 FluoroSpot responses to AdV peptide pools covering: the pentons of human adenoviruses C5 and D26, and the penton of chimpanzee adenovirus Y25 (vector backbone ChAdOx1) as well as a polyclonal anti‐CD3/CD28 antibody T cell stimulation as a positive control of PBMC from healthy blood donors, calculated as spot‐forming units (SFU) per 106 PBMC (background corrected).

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  Figure 4. Frequency of IFNγ (A) and IL2 (B) responses to adenovirus peptide pools covering the hexon proteins of Y25 and C5 in ChAdOx1 vaccine recipients and controls (mRNA vaccine or no vaccine), expressed as spot‐forming units per 106 PBMC (background corrected). IFNγ responses are shown as filled bars. IL‐2 responses are shown as bars with horizontal stripes. Mean individual responses to ChAdV‐Y25 are shown as triangles. Mean individual responses to HAdV‐C5 are shown as circles. Control recipients are shown in purple, ChAdOx1 recipients in blue. P values were calculated with a Mann–Whitney U test.

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  Figure 5. Plots showing the correlation between the frequency of IFNγ responses for pairs of hexons for ten healthy donors. Each axis shows the number of SFUs per 106 PBMC produced by each donor in response to hexon peptide pool stimulation. Spearman's r correlation coefficients and two‐tailed p values are shown for statistically significant correlations.

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  Figure 6. (A) Diagram showing the location of the adenovirus A12 hexon in genomic context, and the composition of the conserved and variable peptide pools derived from this protein. (B) Frequency of PBMC IFNγ responses to the HAdV‐A12 hexon, divided into two conserved and one variable epitope pool. Points show the mean (n = 3) and SEM for each donor and pool. The dotted line indicates a positive IFNγ response was defined as greater than 32.5 SFU/106 PBMC (background corrected). (C) Frequency of PBMC IL2 responses to the conserved and variable epitope pools. The dotted line indicates a positive IL2 response was defined as greater than 17 SFU/106 PBMC (background corrected).