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. Author manuscript; available in PMC: 2014 Jun 14.
Published in final edited form as: J Immunol. 2013 Oct 7;191(10):5124–5138. doi: 10.4049/jimmunol.1301415

FIGURE 6.

FIGURE 6

CD8+ T cell–dependent protective immunity against ocular herpes induced by ASYMP epitopes in humanized HLA transgenic mice. Three groups of age-matched female HLA-A*02:01 transgenic mice (n = 10 each) were immunized s.c. with the ASYMP CD8+ T cell human epitopes (gB342–350 and gB561–569) delivered with the CD4+ T cell PADRE epitope emulsified in CpG1826 adjuvant (ASYMP), with the SYMP CD8+ T cell human epitopes (gB183–191 and gB441–449) delivered with the CD4+ T cell PADRE epitope emulsified in CpG1826 adjuvant (SYMP), or with the CpG1826 adjuvant alone (mock) on days 0 and 21. Two weeks after the final immunization, all animals were challenged ocularly with 2 × 105 PFU of HSV-1 (strain McKrae). (A) The eye disease was detected and scored 2 wk after immunization as described in Materials and Methods. (B) Viral loads were detected on day 7 postinfection in eye swabs, as described in Materials and Methods. (C) Immunized and infected mice were examined for survival in a window of 30 d postchallenge, as described in Materials and Methods. (D) Scattergram and linear regression analysis of mouse survival (%) and HSV-specific CD8+ T cell responses after challenge with HSV-1. Correlation was performed using the Pearson test with two-tailed p value analysis (r2 = 0.7996; p < 0.0001). (E) The protective immunity against ocular herpes induced by the ASYMP CD8+ T cell human epitope immunization is abrogated following depletion of CD8+ T cells, but not of CD4+ T cells. Following the second immunization, and before challenge with HSV-1, mice were injected i.p. with six doses (i.e., 1 every other day) of 100 µl saline containing anti-CD4, anti-CD8, or isotype control. Flow cytometry analysis confirmed a decrease in spleen CD4+ and CD8+ T cells in treated mice to consistently <2% after mAb treatment. The p values compare protection achieved in mAb treated versus untreated mice using the ANOVA test. Immunized, mAb-treated, and infected mice were examined for survival in a window of 30 d after challenge. Results are representative of two independent experiments.