antibodies against ter 119  (R&D Systems)

Journal: The Journal of Clinical Investigation

Article Title: Corticosteroids impair epithelial regeneration in immune-mediated intestinal damage

doi: 10.1172/JCI155880

Figure Lengend Snippet: ( A – G ) B6-into-BALB/c transplantation of BM with or without T cells; recipient mice were treated with MP (2 mg/kg) or vehicle i.p. daily, starting on day 7 through day 14 after BMT. Recipient mice were evaluated on day 14 after BMT. ( B ) CD44 + CD62L – cell proportion of CD45 + CD3 + CD4 + CD8 – splenocytes ( n = 3–5 mice per group). ( C ) Representative images of ileum. Scale bars: 50 μm. ( D ) SI lymphocytic infiltrate histopathology score ( n = 8–9 mice per group). ( E and F ) Ileal crypt frequency and height ( n = 20–28 sections per group). Scale bars: 50 μm. ( G ) Representative images and Ki67 + cell frequency ( n = 7–14 sections per group). ( H and I ) Representative images and B6 SI organoid frequency after culturing with or without anti-CD3/CD28–activated Nr3c1 fl/fl or Nr3c1 fl/fl Cd4-Cre B6 T cells with or without MP (10 μM) for 4 days ( n = 5–6 wells per group). Scale bars: 500 μm. ( J ) B6 SI organoid frequency after in vivo MP (or vehicle) treatment prior to crypt isolation and subsequent culturing with anti-CD3/CD28–activated WT B6 T cells on day 6 of culturing ( n = 12 wells per group). ( K and L ) Representative images and organoid frequency after culturing with or without MP (10 μM) and rmIFN-γ for 6 days ( n = 6 wells per group). Scale bars: 500 μm. ( M and N ) qPCR to determine Bcl2 and Bak1 expression in organoids cultured with or without MP (10 μM) and rmIFN-γ (1 ng/mL) for 3 days ( n = 6 wells per group). ( O and P ) Representative images and human organoid frequency after culturing with or without MP (10 μM) and rhIFN-γ (2 ng/mL) for 7 days ( n = 12 fields of view in 6 wells per group). Scale bars: 1,000 μm. * P < 0.05, ** P < 0.01, and *** P < 0.001, by 2-tailed t test or 1-way ANOVA. Data are representative of at least 2 independent experiments or were combined from 2 experiments ( A – G ).

Article Snippet: Donor T cells were purified from splenocytes using biotin-labeled Abs against CD19 (1D3), CD45R (RA3–6B2), CD11b (M1/70), CD11c (N418), CD49b (DX5), NK1.1 (PK136), TCR γΔ (GL3), and TER-119 (TER-119) from STEMCELL Technologies, followed by streptavidin RapidSpheres depletion with an EasySep magnet, also from STEMCELL Technologies.

Techniques: Transplantation Assay, Histopathology, In Vivo, Isolation, Expressing, Cell Culture