Skin-resident DCs of K14.E7 mice are impaired in their ability to process antigen in vivo.

<p><b>(A)</b> Whole dermal and epidermal single cell suspensions of C57BL/6 and K14.E7 ear skin were incubated with 1ug/ml DQ-OVA for 1 hour and analyzed for photolytic degradation by flow cytometry. DC subsets were gated according to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0152886#pone.0152886.t001" target="_blank">Table 1</a>. The green median fluorescent intensity of whole LCs, DQ-OVA+ CD103+ DCs and DQ-OVA+ CD11b+ DCs was compared. <b>(B)</b> C57BL/6 and K14.E7 mice were injected intradermally into the ear with 20μl of 1mg/ml labeled DQ-OVA. 24 hours later, single cell suspensions of treated and untreated ear dermis and epidermis were prepared and analyzed by flow cytometry for photolytic degradation of applied DQ-OVA, as recognized by fluorochrome unmasking. Histogram overlays of fluorescence in LCs, CD103⁺ dDCs and CD11b⁺ dDCs determined according to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0152886#pone.0152886.t001" target="_blank">Table 1</a> in one representative C57BL/6 (grey line) and K14.E7 (black line) mouse ear treated with DQ-OVA. The change in green median fluorescent intensity from untreated to DQ-OVA treated in dermal and epidermal LCs, CD103⁺ dDCs and CD11b⁺ dDCs from K14E7 (●) and C57BL/6 (■) mouse ears (n = 4) was compared. <b>(C)</b> C57BL/6 and K14.E7 mice were injected intravenously with OT-I CD8+ T cells and immunized intradermally into the ear pinnae with 20μl of 1mg/ml OVA. One week later, IFNγ secretion by OT-I T cells was analyzed by recalling ear-draining lymph node cells with SIINFEKL antigen in an ELISpot assay. Depicted are spots per 2x10⁵ cells. Each data point represents means of triplicates of one individual animal (n = 5).