Therapeutic Prime/Pull Vaccination of HSV-2 Infected Guinea Pigs with the Ribonucleotide Reductase 2 (RR2) Protein and CXCL11 Chemokine Boosts Antiviral Local Tissue-Resident and Effector Memory CD4⁺and CD8⁺T Cells and Protects Against Recurrent Genital Herpes

ABSTRACT Following acute herpes simplex virus type 2 (HSV-2) infection, the virus undergoes latency in sensory neurons of the dorsal root ganglia (DRG). Intermittent virus reactivation from latency and shedding in the vaginal mucosa (VM) causes recurrent genital herpes. While T-cells appear to play a role in controlling virus reactivation and reducing the severity of recurrent genital herpes, the mechanisms for recruiting these T-cells into DRG and VM tissues remain to be fully elucidated. The present study investigates the effect of CXCL9, CXCL10, and CXCL11 T-cell-attracting chemokines on the frequency and function of DRG- and VM-resident CD4 + and CD8 + T cells and its effect on the frequency and severity of recurrent genital herpes. HSV-2 latent-infected guinea pigs were immunized intramuscularly with the HSV-1 RR2 protein (Prime) and subsequently treated intravaginally with the neurotropic adeno-associated virus type 8 (AAV-8) expressing CXCL9, CXCL10, or CXCL11 T-cell-attracting chemokines ( Pull) . Compared to the RR2 therapeutic vaccine alone, the RR2/CXCL11 prime/pull therapeutic vaccine significantly increased the frequencies of functional tissue-resident (T RM cells) and effector (T EM cells) memory CD4 + and CD8 + T cells in both DRG and VM tissues. This was associated with less virus shedding in the healed genital mucosal epithelium and reduced frequency and severity of recurrent genital herpes. These findings confirm the role of local DRG- and VM-resident CD4 + and CD8 + T RM and T EM cells in reducing virus reactivation shedding and the severity of recurrent genital herpes and propose the novel prime/pull vaccine strategy to protect against recurrent genital herpes. IMPORTANCE The present study investigates a novel prime/pull therapeutic vaccine strategy to protect against recurrent genital herpes in the guinea pig model. HSV-2 infected guinea pigs were vaccinated using a recombinantly expressed herpes tegument protein-RR2 ( prime ), followed by intravaginal treatment with the neurotropic adeno-associated virus type 8 (AAV-8) expressing CXCL9, CXCL10, or CXCL11 T-cell-attracting chemokines (pull). The RR2/CXCL11 prime/pull therapeutic vaccine elicited a significant reduction in virus shedding in the vaginal mucosa and decreased the severity and frequency of recurrent genital herpes. This protection was associated with increased frequencies of functional tissue-resident (T RM cells) and effector (T EM cells) memory CD4 + and CD8 + T cells infiltrating latently infected DRG tissues and the healed regions of the vaginal mucosa. These findings shed light on the role of tissue-resident (T RM cells) and effector (T EM cells) memory CD4 + and CD8 + T cells in DRG and vaginal mucosa (VM) tissues in protection against recurrent genital herpes and propose the prime/pull therapeutic vaccine strategy in combating genital herpes. TWEET A therapeutic RR2/CXCL11 prime/pull vaccine reduced recurrent genital herpes more effectively than therapeutic vaccination with a subunit HSV RR2 antigen alone.