Fig. 5: Nociceptors are required to promote a robust CD8 T cell response to HSV-1 infection.

a Immunofluorescence staining of the epidermis of DTA (top panels) and Nav_1.8-DTA (bottom panels) mice. Neurons were stained with anti-PGP9.5 antibodies (in red), Langerhans cells (LC) were stained with anti-langerin antibodies (in green) and T cells were stained with anti-CD3 antibodies (in blue). N = 2–3 mice per group. b Control DTA and Nav_1.8-DTA mice were infected with HSV-OVA-TK⁻ by flank scarification. Skin DC subsets were analysed by flow cytometry using the gating strategy presented in Fig. 3b. Absolute numbers of cDC1, cDC2 and LC on day 6 pi are shown. Each dot represents the data obtained for one mouse. N = 8–11 mice per group. The data are presented as mean ± SEM. P values were obtained by using a Mann–Whitney test (two-tailed). c, d In vitro activation of OT-I T cells. OT-I T cells from naive mice were purified, stained with a fluorescent marker and co-cultured for 48 h with purified CD11c⁺ DCs from infected control DTA (blue) or Nav_1.8-DTA (red) brachial and axillary lymph nodes isolated on 4 days pi. Representative dot plots and quantification of the frequency of proliferating OT-I T cells in the presence (c) or absence (d) of the OVA peptide (SIINFEKL). N = 5–6 per group. The data are presented as mean ± SEM. P values were obtained by using a Mann–Whitney test (two-tailed). e Experimental design used for monitoring antiviral CD8 T cell responses in vivo. Control DTA (blue) and Nav_1.8-DTA (red) recipient mice were injected with 5 × 10⁴ naive virus-specific T cells (CD45.1⁺ OT-I T cells) 1 day before HSV-OVA-TK⁻ infection by flank scarification and were analysed 4 or 8 days pi. f, g Absolute numbers of virus-specific T cells (OT-I) in dLN (n = 4–24 mice per group) (f) and spleens (n = 3–30 per group) (g) were determined at the indicated time points. Virus-specific T cells were detected by flow cytometry and gated based on their expression of the Vα2, CD45.1, CD3 and CD8 markers, the gating strategy used is shown in Supplementary Fig. 4e. The results shown are representative of at least three independent experiments. The data are presented as mean ± SEM; P values were obtained by using a Mann–Whitney test (two-tailed).