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. 2023 Jun 22;14:1183788. doi: 10.3389/fimmu.2023.1183788

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Copyright © 2023 Vietzen, Staber, Berger, Furlano, Kühner, Lubowitzki, Pichler, Strassl, Cornelissen and Puchhammer-Stöckl

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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LMP-1-derived peptides are associated with the development of EBV⁺ lymphomas and inhibit NKG2A⁺ NK cells. (A–D) Distribution of LMP-1 variants in patients with (A) asymptomatic reactivations (N=96), (B) symptomatic reactivations (N=96), (C) EBV⁺HL (N=25), (D) EBV⁺nHL (N=38). Fractions represent the relative frequency of the LMP-1 peptide GGDPHLPTL, GSDPHLPTL, GGDPHLPPL, GGDPPLPTL, GCDPHLPTL, GIDPHLPTL, GAGPHLPTL, GGDTPLPTL, GDDPHLPTL, GGDPHVPTL and GTDPHLPTL variants. ⁺ The frequency of the LMP-1 variants was compared to the asymptomatic cohort by the Chi² Test. (E, F) HLA-E stabilisation assay. (E) HLA-E stabilisation assay: Raji cells were incubated with indicated concentrations of the positive control (VMAPRTLFL) or the LMP-1 peptide-derived GGDPHLPTL, GSDPHLPTL, GGDPHLPPL, GGDPPLPTL, GCDPHLPTL, GIDPHLPTL, GAGPHLPTL, GGDTPLPTL, GDDPHLPTL, GGDPHVPTL and GTDPHLPTL variants. The HLA-E surface expression was then assessed after 16h of co-culture by flow-cytometry. Plots represent the mean ( ± SD) of three independent replicates. Each peptide was compared to the positive control using RM one-way ANOVA (with the Geisser-Greenhouse correction). (F) NKG2A⁺NKG2C^- NK cell inhibition assay: Raji cells were incubated with indicated concentrations of the positive control (VMAPRTLFL) or the LMP-1 peptide-derived GGDPHLPTL, GSDPHLPTL, GGDPHLPPL, GGDPPLPTL, GCDPHLPTL, GIDPHLPTL, GAGPHLPTL, GGDTPLPTL, GDDPHLPTL, GGDPHVPTL and GTDPHLPTL variants and were then co-cultured with pre-activated NKG2A⁺NKG2C^- NK cells from 12 blood donors. Plots represent the mean ( ± SD) of 12 independent replicates. Each peptide was compared to the positive control using RM one-way ANOVA (with the Geisser-Greenhouse correction). p < 0.05 was considered significant. *p < 0.05; **p < 0.01; ***p < 0.001. EBV⁺HL, EBV⁺ Hodgkin lymphoma, EBV⁺nHL, EBV⁺ non-Hodgkin lymphoma, Pos. Ctrl., positive control.