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. 2022 Jun 23;18(6):e1010403. doi: 10.1371/journal.ppat.1010403

Fig 3.

Fig 3

CMV vaccine immunogenicity evaluation in multiple HLA expressing mice: (a) Schematic outline of CMV vaccine immunogenicity evaluation in HLA expressing mice. CMV vaccine formulation included CMVpoly (30μG) and gB (5μg) proteins admixed with CpG1018 (50μg) adjuvant. Five different human HLA expressing mice (HLA A1, A2, A24, B8 and B35) were immunised subcutaneously with CMV vaccine (N = 6) or CpG1018 alone (N = 4) as a placebo control on days 0 and 21 and CMV-specific cellular and humoral immune responses were assessed on day 28. (a and b) Ex vivo CMV-specific CD8+ and CD4+ T cell responses in HLA A1, A2, A24, B8 and B35 expressing mice immunized with CMV vaccine. Splenocytes from immunized mice were stimulated with HLA matched CD8+ T cell epitopes (Panel a) or CMV gB overlapping pepmix (Panel b) and then assessed for IFNg expression using intracellular cytokine assays. (d and e) In vitro expansion of CMV-specific CD8+ and CD4+ T cells from splenocytes of immunized mice following in vitro stimulation with HLA matched CD8+ T cell epitopes (Panel d) or CMV gB overlapping pepmix (Panel e). Splenocytes were stimulated with HLA matched CD8+ CMV T cell epitopes or gB overlapping pepmix and then cultured for 10 days in the presence of IL-2. After incubation these T cells were assessed for IFNg expression using intracellular cytokine assays. Error bars are SEMs. Statistics are indicated in comparison with control group. If statistics are not indicated, then the comparison was not significant (ns). P vales were also calculated between the vaccinated mice. In Fig 3B HLA A24 vs HLA A1 p = 0.61, HLA A24 vs HLA A2 p = 0.029, HLA A24 vs HLA B8 p = 0.06 and HLA A24 vs HLA B35 p = 0.02. In Fig 3C HLA A24 vs HLA A1 p = 0.004, HLA A24 vs HLA A2 p = 0.02, HLA A24 vs HLA B8 p = 0.006 and HLA A24 vs HLA B35 p = 0.01. In Fig 3D HLA A24 vs HLA A1 p = 0.009, HLA A24 vs HLA A2 p = 0.001, HLA A24 vs HLA B8 p = 0.52 and HLA A24 vs HLA B35 p = 0.001. In Fig 3E HLA A24 vs HLA A1 p = 0.009, HLA A24 vs HLA A2 p = 0.006, HLA A24 vs HLA B8 p = 0.003 and HLA A24 vs HLA B35 p = 0.009. * P < 0.05; ** P < 0.01; *** P < 0.001 by Welch’s t test.