Figure 7. TNF-α and IL-1β restore the innate and adaptive immune response of DNCB CHS in K14^Cre/+ NK1R^KO mice.

(A) Innate immune response by percentage of ear thickness 24 hours after sensitization of K14^Cre/+ NK1R^KO (K14-NK1R^KO) mice treated on the right ear with topical DNCB alone or with IL-1β, TNF-α, or both (i.d.). Cre^neg littermates (Litt) were treated with topical DNCB alone. Controls included mice treated with topical vehicle (Veh) plus PBS (i.d.), or topical vehicle and IL-1β plus TNF-α (i.d.). Each circle represents 1 mouse (n = 5). (B) Microscopic images of mouse ears shown in A illustrating epidermal-dermal thickness (vertical lines) and skin-infiltrating leukocytes. Scale bars: 50 μm. (C and D) Quantification (FACS) of skin-migratory dendritic cells (smiDCs) in skin-draining lymph nodes (sdLNs) 72 hours after sensitization as in A. Each circle represents smiDC number per individual sdLN (n = 3). (D) Representative dot plots of the smiDCs in sdLNs. (E) Effector immune response by percentage of ear thickness in K14-NK1R^KO mice sensitized in the abdominal skin with DNCB alone or with IL-1β, TNF-α, or both (i.d.) and elicited in the right ear 5 days later with topical DNCB alone or with IL-1β, TNF-α, or both (i.d.). Cre^neg littermates were sensitized and elicited with DNCB alone. Controls included mice sensitized and elicited with topical vehicle plus PBS (i.d.), or topical vehicle and IL-1β plus TNF-α (i.d.). Each circle represents 1 mouse (n = 5). (F) Microscopic images of the ears of mice in E 48 hours after elicitation, showing epidermal-dermal thickness (vertical lines) and skin-infiltrating leukocytes. Scale bars: 50 μm. (B and F) H&E; original magnification, ×200. Means ± 1 SD were analyzed by 1-way ANOVA and post hoc Student-Newman-Keuls test; **P < 0.01, ***P < 0.001, ****P < 0.0001.