Characterization of Langerin-expressing dendritic cell subsets in the normal cornea

Invest Ophthalmol Vis Sci. 2011 Jun 28;52(7):4598-604. doi: 10.1167/iovs.10-6741.

Abstract

Purpose: In addition to Langerhans cells (LCs), other dendritic cells (CD11c(+)) have recently been shown to express Langerin (c-type lectin). In skin, (non-LC) Langerin+ dendritic cells initiate adaptive immunity. However, whether such dendritic cells (DC) reside in the cornea, an immune-privileged tissue, is unknown.

Methods: Normal C57BL/6 corneas were harvested for qRT-PCR analyses of Langerin expression in the epithelium versus stroma. Immunohistochemistry for Langerin was also performed. Single-cell preparations of epithelium versus stroma were FACS analyzed for CD11c, CD11b, and CD103 expression. Fluorescence microscopy of corneas from muLangerin-eGFP mice (in which all CD11c(+) Langerin+ cells express eGFP), huLangerin-DTA mice (only LCs are constitutively deleted), and huLangerin-Cre eYFP-flox (only LCs express eYFP) was performed.

Results: qRT-PCR, immunohistochemistry, and FACS analysis identified CD11c(+) Langerin+ cells in the epithelium and stroma. Similarly, corneas of muLangerin-eGFP mice contained eGFP+ cells in the epithelium and stroma. However, FACS analysis indicated phenotypically differing CD11c(+) Langerin+ populations in the epithelium (CD11b(low)CD103(low)) versus stroma (CD11b(+)CD103(low)). Additionally, corneas from huLangerin-DTA mice were devoid of Langerin+ cells in the epithelium but were detectable in the stroma. In corneas from huLangerin-Cre eYFP-flox, eYFP+ cells were detectable in the epithelium but not in the stroma.

Conclusions: The normal corneal epithelium is endowed with CD11c(+) Langerin+ cells that are LCs, whereas the stroma is endowed with a separate population of (non-LC) Langerin+ DCs. These findings should henceforth facilitate the examination of Langerin-expressing DC subsets in the immunopathogeneses of conditions such as keratoconjunctivitis sicca, allergic keratoconjunctivitis, and corneal allograft rejection.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antigens, Surface / biosynthesis
  • Antigens, Surface / genetics*
  • Cornea / cytology*
  • Cornea / metabolism
  • Corneal Stroma / cytology
  • Corneal Stroma / metabolism
  • Dendritic Cells / cytology
  • Dendritic Cells / metabolism*
  • Flow Cytometry
  • Gene Expression Regulation*
  • Immunohistochemistry
  • Lectins, C-Type / biosynthesis
  • Lectins, C-Type / genetics*
  • Male
  • Mannose-Binding Lectins / biosynthesis
  • Mannose-Binding Lectins / genetics*
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics*

Substances

  • Antigens, Surface
  • Cd207 protein, mouse
  • Lectins, C-Type
  • Mannose-Binding Lectins
  • RNA, Messenger