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Peptide epitope immunoreactivity in peripheral blood mononuclear cells (PBMCs) isolated from individuals naïve to SARS‐CoV‐2, as well as following vaccination and natural infection. Interferon gamma (IFN‐γ) expression quantified through a high‐throughput RT‐qPCR (HTS‐RT‐qPCR) assay with fold change (ΔΔ C t ) determined relative to the endogenous control reference gene Ribosomal Protein L13a (RPL13a) of PBMCs isolated from 12 individuals naïve to SARS‐CoV‐2 S‐protein (naïve; white dots), following homologous AstraZeneca COVID‐19 vaccination (vaccinated; grey dots), and following infection with SARS‐CoV‐2 (naturally infected; black dots) stimulated with 170 SARS‐CoV‐2 S‐protein peptide epitopes. Shown are data sorted by donor (a) , sorted into HLA‐A2, HLA‐A3/11, HLA‐A24, HLA‐B7, HLA‐B8, other and Class II supertype classifications (b) , and data showing matched peptide kinetics tracking immunoreactive epitopes (ΔΔ C t > 2) across the naïve, vaccinated and naturally infected (c) . Epitopes with ΔΔ C t between 0 and 10 shown graphically while all data were considered for statistical analysis. Selected immunoreactive peptides shown with three‐letter codes.

Journal: Clinical & Translational Immunology

Article Title: Differential reactivity of SARS ‐ C o V ‐2 S‐protein T‐cell epitopes in vaccinated versus naturally infected individuals

doi: 10.1002/cti2.70031

Figure Lengend Snippet: Peptide epitope immunoreactivity in peripheral blood mononuclear cells (PBMCs) isolated from individuals naïve to SARS‐CoV‐2, as well as following vaccination and natural infection. Interferon gamma (IFN‐γ) expression quantified through a high‐throughput RT‐qPCR (HTS‐RT‐qPCR) assay with fold change (ΔΔ C t ) determined relative to the endogenous control reference gene Ribosomal Protein L13a (RPL13a) of PBMCs isolated from 12 individuals naïve to SARS‐CoV‐2 S‐protein (naïve; white dots), following homologous AstraZeneca COVID‐19 vaccination (vaccinated; grey dots), and following infection with SARS‐CoV‐2 (naturally infected; black dots) stimulated with 170 SARS‐CoV‐2 S‐protein peptide epitopes. Shown are data sorted by donor (a) , sorted into HLA‐A2, HLA‐A3/11, HLA‐A24, HLA‐B7, HLA‐B8, other and Class II supertype classifications (b) , and data showing matched peptide kinetics tracking immunoreactive epitopes (ΔΔ C t > 2) across the naïve, vaccinated and naturally infected (c) . Epitopes with ΔΔ C t between 0 and 10 shown graphically while all data were considered for statistical analysis. Selected immunoreactive peptides shown with three‐letter codes.

Article Snippet: T‐cell peptide epitopes from SARS‐CoV‐2 S‐protein were synthesised at 95% purity (Mimotopes, Melbourne, Australia) and resuspended in dimethyl sulfoxide (DMSO) at a concentration of 20 mg mL −1 .

Techniques: Isolation, Infection, Expressing, High Throughput Screening Assay, Quantitative RT-PCR, Control