Journal: Journal of Translational Medicine

Article Title: HLA-E-restricted SARS-CoV-2 epitopes drive CD8 + T cell memory in convalescent and vaccinated individuals: implications for the design of next-generation vaccines and immunotherapeutics

doi: 10.1186/s12967-026-07825-8

Figure Lengend Snippet: Representation of the combined cytokine expression results for a discovery cohort of 7 convalescent COVID-19 patients. The panel A shows the frequency of the CD8 + T cell cytokine (IFN-γ, TNF-α, and IL-2) response to K562/HLA-E cells pulsed with HLA-E-binding SARS-CoV-2 peptides. White boxes, no cytokine response (the total percentage of cytokine + CD8 + T cells is < 0.01); light gray boxes, the total percentage of cytokine + CD8 + T cells ranges from 0.01- 0.2%; gray boxes, the total percentage of cytokine + CD8 + T cells ranges from 0.4- 0.6%; dark gray boxes, the total percentage of cytokine + CD8 + T cells ranges from 0.6- 0.8% and black boxes, the total percentage of cytokine + CD8 + T cells ranges from 0.8–1% or more. Panels B , C , and D show the frequency of CD8 + T cells positive for IFN-γ, TNF-α, and IL-2 respectively, upon stimulation with K562/HLA-E cells pulsed with single peptide

Article Snippet: Intracellular cytokine staining was performed using fluorochrome-conjugated antibodies against IFN-γ FITC (REA600), TNF-α PE (REA656), and IL-2 PE-Vio 770 (REA689) (Miltenyi Biotec) in permeabilization buffer.

Techniques: Expressing, Binding Assay

Journal: Journal of Translational Medicine

Article Title: HLA-E-restricted SARS-CoV-2 epitopes drive CD8 + T cell memory in convalescent and vaccinated individuals: implications for the design of next-generation vaccines and immunotherapeutics

doi: 10.1186/s12967-026-07825-8

Figure Lengend Snippet: Frequencies of HLA-E/SARS-CoV-2 peptide-specific CD8 + T cells in convalescent and hospitalized severe COVID-19 patients. A) Frequency of IFN-γ, TNF-α, and IL-2 positive CD8 + T cells in convalescent (n.=21) and hospitalized severe COVID-19 patients (n.=17) upon stimulation with the pool of SARS-CoV-2 peptides. Unstimulated cells were used as a negative control (NC), while the cells stimulated with PepTivator® were considered a positive control of SARS-CoV-2-specific response. The graph represents the median and the error bars. Two-way ANOVA or mixed models were used to compare the three experimental points. *p<0.05; **p<0.01. B) Frequency of IFN-γ, TNF-α and IL-2 positive CD8 + T cells in convalescent and hospitalized severe COVID-19 patients upon stimulation with the pool of HLA-E-SARS-CoV-2 peptides. Unstimulated cells were used as a negative control (NC), while the cells stimulated with PepTivator® were considered as a positive control of SARS-CoV-2-specific response. The graph represents the median and the error bars. Two-way ANOVA or mixed models were used to compare the three experimental points. *p<0.05. C) Frequency of IFN-γ, TNF-α and IL-2 positive CD8 + T cells in vaccinated subjects (n.=10) upon stimulation with SP09 peptide. Unstimulated cells were used as a negative control (NC), while the cells stimulated with PepTivator® were considered as a positive control of SARS-CoV-2-specific response. The graph represents the median and the error bars. The Kruskal-Wallis test was used to compare the values of the groups. *p ≤ 0.05, ***p≤0.001

Article Snippet: Intracellular cytokine staining was performed using fluorochrome-conjugated antibodies against IFN-γ FITC (REA600), TNF-α PE (REA656), and IL-2 PE-Vio 770 (REA689) (Miltenyi Biotec) in permeabilization buffer.

Techniques: Negative Control, Positive Control

Journal: Journal of Translational Medicine

Article Title: HLA-E-restricted SARS-CoV-2 epitopes drive CD8 + T cell memory in convalescent and vaccinated individuals: implications for the design of next-generation vaccines and immunotherapeutics

doi: 10.1186/s12967-026-07825-8

Figure Lengend Snippet: Cytokine responses of CD8 + T cells stimulated with peptide SP09, in vaccinated individuals, with or without subsequent infection. A ) frequency of IFN-γ, TNF-α, and IL-2 positive CD8 + T cells after subtraction of the respective NC in 15 vaccinated subjects with two doses, and 10 with three doses. The graph represents the median and the error bars. The Mann-Whitney test was used to compare the values of the groups. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. B ) comparison between healthy individuals vaccinated with three doses (n. = 3) of the mRNA BNT162b2 vaccine without infection and individuals vaccinated with three vaccine doses who got infected with SARS-CoV-2 (n. = 7). The Mann-Whitney test was used to compare the values of the groups * p ≤ 0.05, **** p ≤ 0.0001

Article Snippet: Intracellular cytokine staining was performed using fluorochrome-conjugated antibodies against IFN-γ FITC (REA600), TNF-α PE (REA656), and IL-2 PE-Vio 770 (REA689) (Miltenyi Biotec) in permeabilization buffer.

Techniques: Infection, MANN-WHITNEY, Comparison