Heterologous prime-boost vaccination protects against EBV antigen–expressing lymphomas
Julia Rühl, … , Carol S. Leung, Christian Münz
Julia Rühl, … , Carol S. Leung, Christian Münz
Published March 12, 2019
Citation Information: J Clin Invest. 2019;129(5):2071-2087. https://doi.org/10.1172/JCI125364.
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Research Article Immunology

Heterologous prime-boost vaccination protects against EBV antigen–expressing lymphomas

Abstract

The Epstein-Barr virus (EBV) is one of the predominant tumor viruses in humans, but so far no therapeutic or prophylactic vaccination against this transforming pathogen is available. We demonstrated that heterologous prime-boost vaccination with the nuclear antigen 1 of EBV (EBNA1), either targeted to the DEC205 receptor on DCs or expressed from a recombinant modified vaccinia virus Ankara (MVA) vector, improved priming of antigen-specific CD4+ T cell help. This help supported the expansion and maintenance of EBNA1-specific CD8+ T cells that are most efficiently primed by recombinant adenoviruses that encode EBNA1. These combined CD4+ and CD8+ T cell responses protected against EBNA1-expressing T and B cell lymphomas, including lymphoproliferations that emerged spontaneously after EBNA1 expression. In particular, the heterologous EBNA1-expressing adenovirus, boosted by EBNA1-encoding MVA vaccination, demonstrated protection as a prophylactic and therapeutic treatment for the respective lymphoma challenges. Our study shows that such heterologous prime-boost vaccinations against EBV-associated malignancies as well as symptomatic primary EBV infection should be further explored for clinical development.

Authors

Julia Rühl, Carmen Citterio, Christine Engelmann, Tracey Haigh, Andrzej Dzionek, Johannes Dreyer, Rajiv Khanna, Graham S. Taylor, Joanna B. Wilson, Carol S. Leung, Christian Münz

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Figure 1

Human CD4+ and CD8+ T cell recognition of EBNA1 carrying or encoding vaccine formulations.

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Human CD4+ and CD8+ T cell recognition of EBNA1 carrying or encoding vac...
(A) Structure of humanized EBNA1-Ab fusion proteins. (B) Western blot analysis of human αDEC205-EBNA1 Ab under reducing conditions using rat αEBNA1 Ab (clone 1H4). Lane 1 represents heavy-chain EBNA1 (100 kDa) and lane 2 recombinant truncated EBNA1. (C) Western blot analysis of viral vectors encoding truncated EBNA1, using rat αEBNA1 Ab (clone 1H4) 48 hours after infection of HEK293T cells. MVA-E1 carries EBNA1 without the Gly/Ala repeat and runs at approximately 45 kDa (25); MVA-IiE1 has the additional invariant chain domain. Lenti-E1 carries only EBNA1 from aa 400–641 with an approximate size of 30 kDa. Infection with Adeno–E1-LMP also leads to expression of the Gly/Ala repeat–deleted EBNA1 protein, however with additional LMP polyepitopes (26), and migrates at approximately 60 kDa. Lane 6 represents uninfected HEK293T cells and lane 7 recombinant truncated EBNA1. (D and E) Autologous PBMCs were incubated with medium for 4 hours with 1 μg/ml EBNA1 fused to an Ab against the indicated receptors, or for 1 hour with the cognate peptides for the respective T cell clones. Coculture with (D) EBNA1-specific CD4+ T cell clones, with cognate epitope NLR and SNP shown in the gray bars, and (E) EBNA1-specific CD8+ T cell clones, with cognate epitope HPV shown in the white bars. T cell activity was measured by IFN-γ release into the supernatant. IFN-γ signal is shown as a percentage of the peptide control. Data are shown as the mean ± SD of at least 2 independent experiments. ***P < 0.005 versus unspecific CD207-targeting; 1-way ANOVA with Bonferroni’s pre-test . (F and G) Autologous PBMCs were infected with DMSO control, MVA-EBNA1, MVA-liEBNA1, or Adeno–EBNA1-LMP at a MOI of 10 for 48 hours and with Lenti-EBNA1 or Lenti-IiEBNA1 for 96 hours. Coculture with (F) EBNA1-specific CD4+ T cell clones, with cognate epitope NLR and SNP shown in the light gray bars and cognate epitope AEG shown in the dark gray bars, and (G) EBNA1-specific CD8+ T cell clones, with cognate epitope HPV shown in the white bars. T cell activity was determined as in D and E. Data are shown as the mean ± SD of 2 independent experiments. **P < 0.01 and ***P < 0.005; 1-way ANOVA plus Bonferroni’s pre-test.