Anti il 7

Draining lymph nodes (DLN) and conjunctivae were collected from DED mice and cultured in RPMI (Invitrogen) supplemented with 10% FBS. Alternatively, single cell suspensions were prepared from DLN and CD4⁺ T cells were enriched using the negative isolation kit (Miltenyi Biotec). Subsequently, the CD44^hiCD62L⁻ subpopulations were further sorted using MoFlo FACS sorter (Dako Cytomation). The tissue explants or CD44^hiCD62L⁻CD4⁺ cells were treated with anti-IL-7 (10 μg/ml, R&D Systems), anti-IL-15 (5 μg/ml, eBioscience), anti-IL-7 (10 μg/ml) + anti-IL-15 (5 μg/ml), anti-IL-7Rα (10 μg/ml, R&D Systems) + anti-IL-15Rα (10 μg/ml, R&D Systems) Abs, IL-7 (20 ng/ml, PeproTech), IL-15 (20 ng/ml, PeproTech), or IL-7 (20 ng/ml) + IL-15 (20 ng/ml) for 72 hours. Memory Th17 cells were then examined by flow cytometry.