Figure 2.
BZLF1-dervied SQAPLPCVL and HLA-E–restricted CD8+T-cell responses prevent the development of IM. (A-B) Analysis of SQAPLPCVL-specific and HLA-E–restricted CD8+ T-cell responses, evaluated between 12 healthy EBV-seropositive individuals and 6 healthy EBV-seronegative blood donors by flow cytometry. Enriched CD8+ T cells were stimulated with K562-HLA-E∗0103/0103 cells, K562-HLA-E∗0101/0101 cells, or K562 cells or without cells and either 300 μM of the SQAPLPCVL peptide, 300 μM of the LMP-1 peptide pool (consisting of equal concentrations of 11 LMP-1 peptide variants), or without peptides. (C-D) Evaluation of the viral spread and the activation of SQAPLPCVL-specific, HLA-E–restricted CD8+ T cells against EBV-infected K562-CR2-HLA-E∗0103/0103 cells or K562-CR2-HLA-E∗0101/0101 cells. EBV-infected K562-CR2-HLA-E∗0103/0103 cells or EBV-infected K562-CR2-HLA-E∗0101/0101 cells were cultured either alone or together with SQAPLPCVL-specific, HLA-E–restricted CD8+ T cells from 12 healthy EBV-seropositive individuals. (C) K562-CR2-HLA-E∗0103/0103 or K562-CR2-HLA-E∗0101/0101 cells were analyzed for the expression of EBV-BZLF1 after 3, 6, 9, 12, and 15 days after infections by flow cytometry. (D) CD8+ T cells were then analyzed for the expression of IFN-γ after 3, 6, 9, 12, and 15 days after infections by flow cytometry. (A-D) Repeated measures 1-way analysis of variance (with the Geisser-Greenhouse correction) was used to compare differences between the respective groups. Plots represent the mean (±SD) of 12 independent biological replicates. (E) Analysis of proliferating SQAPLPCVL-specific and HLA-E–restricted CD8+ T cells. Enriched and CFSE-stained CD8+ T cells from 12 healthy EBV-seropositive individuals were cocultured together with SQAPLPCVL-peptide pulsed K562-HLA-E∗0103/0103 or K562-HLA-E∗0101/0101 cells. The percentage of proliferating (CFSElow) CD8+ T cells was analyzed by flow cytometry and compared by the Wilcoxon signed-rank test. (F-G) Percentage of SQAPLPCVL-specific and HLA-E–restricted CD8+ T cells in patients with a recent IM (1-8 years; HLA-E∗0101/0101: N = 3; HLA-E∗0101/0103: N = 3; HLA-E∗0103/0103: N = 4) or past IM (10-23 years; HLA-E∗0101/0101: N = 3; HLA-E∗0101/0103: N = 3; HLA-E∗0103/0103: N = 4) or asymptomatic EBV-infected individuals (HLA-E∗0101/0101: N = 7; HLA-E∗0101/0103: N = 7; HLA-E∗0103/0103: N = 6). The percentage of SQAPLPCVL-specific and HLA-E–restricted CD8+ T cells was compared between the groups by the Kruskal-Wallis test and the Dunn posttest. P < .05 was considered significant. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001. IFN-γ, interferon gamma; neg., negative; ns, not significant; pos., positive; w/o, without.

BZLF1-dervied SQAPLPCVL and HLA-E–restricted CD8+T-cell responses prevent the development of IM. (A-B) Analysis of SQAPLPCVL-specific and HLA-E–restricted CD8+ T-cell responses, evaluated between 12 healthy EBV-seropositive individuals and 6 healthy EBV-seronegative blood donors by flow cytometry. Enriched CD8+ T cells were stimulated with K562-HLA-E∗0103/0103 cells, K562-HLA-E∗0101/0101 cells, or K562 cells or without cells and either 300 μM of the SQAPLPCVL peptide, 300 μM of the LMP-1 peptide pool (consisting of equal concentrations of 11 LMP-1 peptide variants), or without peptides. (C-D) Evaluation of the viral spread and the activation of SQAPLPCVL-specific, HLA-E–restricted CD8+ T cells against EBV-infected K562-CR2-HLA-E∗0103/0103 cells or K562-CR2-HLA-E∗0101/0101 cells. EBV-infected K562-CR2-HLA-E∗0103/0103 cells or EBV-infected K562-CR2-HLA-E∗0101/0101 cells were cultured either alone or together with SQAPLPCVL-specific, HLA-E–restricted CD8+ T cells from 12 healthy EBV-seropositive individuals. (C) K562-CR2-HLA-E∗0103/0103 or K562-CR2-HLA-E∗0101/0101 cells were analyzed for the expression of EBV-BZLF1 after 3, 6, 9, 12, and 15 days after infections by flow cytometry. (D) CD8+ T cells were then analyzed for the expression of IFN-γ after 3, 6, 9, 12, and 15 days after infections by flow cytometry. (A-D) Repeated measures 1-way analysis of variance (with the Geisser-Greenhouse correction) was used to compare differences between the respective groups. Plots represent the mean (±SD) of 12 independent biological replicates. (E) Analysis of proliferating SQAPLPCVL-specific and HLA-E–restricted CD8+ T cells. Enriched and CFSE-stained CD8+ T cells from 12 healthy EBV-seropositive individuals were cocultured together with SQAPLPCVL-peptide pulsed K562-HLA-E∗0103/0103 or K562-HLA-E∗0101/0101 cells. The percentage of proliferating (CFSElow) CD8+ T cells was analyzed by flow cytometry and compared by the Wilcoxon signed-rank test. (F-G) Percentage of SQAPLPCVL-specific and HLA-E–restricted CD8+ T cells in patients with a recent IM (1-8 years; HLA-E∗0101/0101: N = 3; HLA-E∗0101/0103: N = 3; HLA-E∗0103/0103: N = 4) or past IM (10-23 years; HLA-E∗0101/0101: N = 3; HLA-E∗0101/0103: N = 3; HLA-E∗0103/0103: N = 4) or asymptomatic EBV-infected individuals (HLA-E∗0101/0101: N = 7; HLA-E∗0101/0103: N = 7; HLA-E∗0103/0103: N = 6). The percentage of SQAPLPCVL-specific and HLA-E–restricted CD8+ T cells was compared between the groups by the Kruskal-Wallis test and the Dunn posttest. P < .05 was considered significant. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001. IFN-γ, interferon gamma; neg., negative; ns, not significant; pos., positive; w/o, without.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal