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. 2018 Jul 31;92(16):e00535-18. doi: 10.1128/JVI.00535-18

FIG 3.

FIG 3

A prime/pull immunization regimen bolsters the function of HSV epitope-specific CD8+ T cells in HLA Tg rabbits. (A) Illustration of the prime/pull immunization regimen and immunology studies in HLA Tg rabbits. HLA Tg rabbits (n = 12) were primed twice, on day 0 and day 15, with a mixture of the UL9196–204, UL25572–580, and UL44400–408 peptides in CpG (ODN2007) adjuvant. On day 29 (14 days after the final peptide immunization), half of the rabbits (n = 6) were left untreated and the other half (n = 6) received a topical ocular treatment with 1010 GC/eye of the AAV8 vector expressing CXCL10. (B) Schematic of the AAV8 vector construct expressing the CXCL10 chemokine and GFP, both under the control of the neurotropic CamKIIα promoter. The frequencies of CD8+ T cells that expressed IFN-γ (C and D) and CD107a/b (E and F) were determined by FACS analysis of cells from the cornea and TG of immunized AAV8-CXCL10-treated and untreated HLA Tg rabbits 10 days after treatment, as described in Materials and Methods. (C) Representative contour plots of IFN-γ+ CD8+ T cells specific to the UL9196–204, UL25572–580, and UL44400–408 epitopes detected in the cornea (COR) (left panels) and TG (right panels) for immunized/untreated, immunized/rAAV8-CXCL10-treated, and mock-immunized rabbits. (D) Average frequencies of IFN-γ+ CD8+ T cells specific to the UL9196–204, UL25572–580, and UL44400–408 epitopes detected in the cornea (top) and TG (bottom) for six immunized/untreated, six immunized/rAAV8-CXCL10-treated, and six mock-immunized rabbits. (E) Representative contour plots of CD107a/b+ CD8+ T cells specific to the UL9196–204, UL25572–580, and UL44400–408 epitopes detected in the cornea (left panels) and TG (right panels) for immunized/untreated, immunized/rAAV8-CXCL10-treated, and mock-immunized rabbits. (F) Average frequencies of CD107a/b+ CD8+ T cells specific to the UL9196–204, UL25572–580, and UL44400–408 epitopes detected in the cornea (top) and TG (bottom) for six immunized/untreated, six immunized/rAAV8-CXCL10-treated, and six mock-immunized rabbits. The results are representative of two independent experiments.