Journal: Mucosal immunology
Article Title: Homeostatic control of conjunctival mucosal goblet cells by NKT-derived IL-13
Figure Lengend Snippet: Effects of CsA treatment on NK and NKT cells. ( a ) Mean±s.d. of conjunctival goblet cell (GC) density in nonstressed (NS) C57BL/6 control mice and mice subjected to desiccating stress for 5 or 10 days (DS5 and DS10, respectively) or DS5 treated with Cyclosporine A (DS5 + CsA) or DS5 treated with CsA vehicle (DS5 + Veh). ( b ) mRNA transcript levels of MUC5AC in conjunctiva of NS C57BL/6 control mice and mice subjected to DS5 and DS10, respectively. ( c ) Mean±.d. of protein concentrations of MUC5AC in conjunctiva of NS C57BL/6 control mice and mice subjected to DS5 and DS10, respectively. ( d, e ) Representative flow cytometry analysis of freshly isolated ocular surface cells stained with pan-NK marker DX5 –fluorescein isothiocyanate (FITC)-conjugated antibody in NS C57BL/6 controls mice and mice subjected to DS5 and DS10, respectively ( d ) or DS5 treated with CsA (DS5 + CsA) or DS5 treated with CsA vehicle (DS5 + Veh) ( e ). Lymphocytes were gated based on characteristic light-scatter properties, and single lymphocytes were gated based on forward scatter height vs. forward scatter area (FSC-A). Numbers in the quadrants indicate the percentage of cells. ( f ) Mean±s.d. of the percentage of DX5+ cells evaluated by flow cytometry in three independent experiments. ( g ) Percentage change in the level of expression of interferon-γ (IFN-γ), IL-13, IL-17A, IL-21, and IL-22 mRNA transcripts in CsA-treated NK + and NK − cell populations, compared with vehicle. *** P
Article Snippet: To evaluate if IL-13 could rescue DS-induced GC loss, C57BL/6 mice were divided into three treatment groups: (i) DS5 control subjects, which received no ocular injections; (ii) vehicle control animals, which received bilateral subconjunctival injections (20μl per eye) of 0.1% BSA in phosphate-buffered saline (DS5 + BSA); (iii) DS5 + IL-13 mice, which received daily bilateral subconjunctival injections of recombinant murine IL-13 (20 ng per eye per injection, dissolved in 20 μl of 0.1% BSA in phosphate-buffered saline (R & D Systems) for 4 days.
Techniques: Mouse Assay, Flow Cytometry, Cytometry, Isolation, Staining, Marker, Expressing