Expansion of Purified ILC2s with MLR and IL-2

Purified ILC2s were sorted into 96-well plates at the density of 100 cells/well and resuspended in mixed lymphocyte reaction (MLR) of gamma-irradiated peripheral blood mononuclear cells (PBMCs) from three healthy volunteers (2 × 10⁶ cells/ml) coupled with 100 IU/ml IL-2 in RPMI 1640 (Sigma-Aldrich) supplemented with 10% heat inactivated human serum and 2 mM l-glutamine and 10 ml/liter penicillin-streptomycin (Sigma-Aldrich; R10H). Every 2–3 d thereafter, half of the medium was replaced with equal volumes of R10H and 100 IU IL-2. Every 4 wk, the cultures were fed with MLR mixed with IL-2.

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Salimi M., Barlow J.L., Saunders S.P., Xue L., Gutowska-Owsiak D., Wang X., Huang L.C., Johnson D., Scanlon S.T., McKenzie A.N., Fallon P.G, & Ogg G.S. (2013). A role for IL-25 and IL-33–driven type-2 innate lymphoid cells in atopic dermatitis. The Journal of Experimental Medicine, 210(13), 2939-2950.

Publication 2013

Cells Glutamine Healthy volunteers Human Irradiated gamma Mixed lymphocyte reaction Penicillin Peripheral blood mononuclear cells Serum Streptomycin

Corresponding Organization :

Other organizations : Medical Research Council, MRC Human Immunology Unit, National Institute for Health Research, University of Oxford, MRC Laboratory of Molecular Biology, University of Cambridge, Trinity College Dublin, Air Force Medical University, John Radcliffe Hospital, National Health Service, St. James's Hospital, National Children’s Research Centre

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Variable analysis

independent variables

Density of purified ILC2s (100 cells/well)
Gamma-irradiated peripheral blood mononuclear cells (PBMCs) from three healthy volunteers (2 × 10^6 cells/ml)
Concentration of IL-2 (100 IU/ml)

dependent variables

Proliferation and expansion of ILC2s

control variables

RPMI 1640 (Sigma-Aldrich) medium supplemented with 10% heat inactivated human serum, 2 mM L-glutamine, and 10 ml/liter penicillin-streptomycin (Sigma-Aldrich; R10H)
Replacement of half the medium with equal volumes of R10H and 100 IU IL-2 every 2-3 days
Feeding of cultures with MLR mixed with IL-2 every 4 weeks

controls

Positive control: None specified
Negative control: None specified

Annotations

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