Journal: Journal of Hepatology

Article Title: Biliary epithelium and liver B cells exposed to bacteria activate intrahepatic MAIT cells through MR1

doi: 10.1016/j.jhep.2015.12.017

Figure Lengend Snippet: MR1-dependent activation of blood and liver MAIT cells by professional and non-professional antigen presenting cells (APC) exposed to bacteria. APC exposed to E. coli were co-cultured with MAIT cells and their activation examined by flow cytometry for CD107a, IFN-γ, TNF-α, CD40L. Dependency on MR1 or IL-12 and IL-18 was assessed by antibody blocking. Activation of blood MAIT by blood macrophages (A) and activation of liver MAIT by THP1 cells (B) and autologous intrahepatic B cells (C) were analyzed. Data are mean ± SEM (A) and median ± interquartile range (B and C). In overlays: untreated (shaded); E. coli (solid line); E. coli + αMR1 (dotted line). ∗ p <0.05; ∗∗∗ p <0.001 by paired t test. UT = untreated.

Article Snippet: CD8 + T cells isolated from blood using CD8 Microbeads (Miltenyi Biotec) or CD3 + T cells isolated from liver by flow sorting were cultured with the E. coli- exposed APCs, in the presence of anti-CD107a (Pe or PeCy5) and blocking antibodies against IL-12p40/70 (5 μg/ml, C8.6, eBioscience), IL-18 (5 μg/ml, 125-2H, MBL International, USA) and MR1 (10 μg/ml) as indicated.

Techniques: Activation Assay, Cell Culture, Flow Cytometry, Blocking Assay

Journal: Journal of Hepatology

Article Title: Biliary epithelium and liver B cells exposed to bacteria activate intrahepatic MAIT cells through MR1

doi: 10.1016/j.jhep.2015.12.017

Figure Lengend Snippet: Bacterially-exposed primary human biliary epithelial cells activate both blood and liver MAIT cells in an MR1 dependent manner. Biliary epithelial cells exposed to E. coli were co-cultured with sorted blood CD8 + (A and B) or liver CD3 + (C) T cells and activation of CD161 ++ Va7.2 + MAIT cells examined by flow cytometry for CD107a, IFNγ-, TNFα, CD40L. Dependency on MR1 or cytokines IL-12 and IL-18 was assessed using function-blocking antibodies. Data are mean ± SEM (A and B) and median ± interquartile range (C). In overlays: untreated (shaded); E. coli (solid line); E. coli + αMR1 (dotted line). ∗ p <0.05; ∗∗∗ p <0.001 by t test. UT = untreated.

Article Snippet: CD8 + T cells isolated from blood using CD8 Microbeads (Miltenyi Biotec) or CD3 + T cells isolated from liver by flow sorting were cultured with the E. coli- exposed APCs, in the presence of anti-CD107a (Pe or PeCy5) and blocking antibodies against IL-12p40/70 (5 μg/ml, C8.6, eBioscience), IL-18 (5 μg/ml, 125-2H, MBL International, USA) and MR1 (10 μg/ml) as indicated.

Techniques: Cell Culture, Activation Assay, Flow Cytometry, Blocking Assay