Abstract Chronic HSV infections of the natural human host are characterized by lifelong latency in dorsal root ganglia. It is not clear which viral proteins can access antigen presentation pathways in the human ganglia. HSV-1-infected ganglia showed increased inflammation compared to virus-negative ganglia. The leukocyte infiltrate in human HSV-infected TG was shown to include HSV-specific CD4 and CD8 T-cells using interferon-gamma (IFN-g) readouts. By cloning HSV-1 ORFeome in a large set of eGFP fusion plasmids, We have interrogated the fine specificity of TG-derived T-cell lines. Antigen presenting cells for CD8 assays were HLA class I heavy chain /HSV-1 ORF co-transfectants, while HLA class II-matched PBMC were used as APC to stimulate CD4 responses. CD8 recognition of discrete HLA class I-restricted epitopes was documented for two distinct HSV-1 glycoproteins, gL and gK, expressed with late kinetics, as well as for the major transactivator tegument protein, VP16. Tetramer and IFN-g staining showed that up to 20% of TG CD8 cells recognized single HSV-1 epitopes. Glycoprotein-specific CD8 T-cells displayed nanomolar avidity and cross-recognized the homologous HSV-2 nonamer peptides. Interestingly, VP16 was also recognized by TG-infiltrating CD4 T-cells. HSV-1 lytic cycle proteins can access ganglionic HLA class I and II antigen presentation pathways during chronic TG infection, implying that local APC somehow sample viral proteins from neuronal reactivation events. Local antigen-specific T-cells may contribute to the life-long equilibrium between latency and clinical reactivation, and influence disease phenotype. Supported by NIH grant AI50132.