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. 2011 Feb 14;208(2):227–234. doi: 10.1084/jem.20101459

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© 2011 Bigley et al.

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Figure 3. — Depletion of tissue DCs, but preservation of LCs and macrophages. (A) Analysis of dermal DC populations by flow cytometry of collagenase-digested dermis. Plots show equivalent number of total cells analyzed (40,000). Gated CD45⁺ HLA-DR⁺ cells normally comprise two fractions separable by autofluorescence (AF) and side scatter (SSC). AF⁻ SSC^low cells (gate 1) contain CD14⁺ DCs, CD1a⁺ DCs, and occasional CD1a^high langerin⁺ migrating LCs (gate 3); AF⁺ SSC^high cells are macrophages (gate 2). CD1a^high cells express langerin (subject 1, inset). Subjects 1 and 3 were analyzed twice, independently; subjects 2 and 4 were analyzed once. (B) Dermal DCs, LCs, and macrophage counts relative to normal controls (n = 22; mean ± SD). (C) Whole-mount immunofluorescence staining for CD1a of epidermal sheets showing representative fields of each subject. (D) Total LC counts averaged from entire low-power image relative to controls (n = 12; mean ± SD). (E) Proportion of Ki-67⁺ LCs in whole-mount epidermal sheets relative to controls (n = 3).