Monocytes from HCs and patients with cirrhosis expressing high (>20%/450 MFI) versus low (<7.5%/300 MFI) AXL levels were isolated using the Pan Monocyte Isolation Kit (Miltenyi Biotec) and co-cultured with allogeneic CD3+ T cells from a different healthy donor, isolated using the Pan T Cell Isolation Kit (Miltenyi Biotec), in a 1:1 ratio. T cell stimulation was induced with anti-CD2/CD3/CD28 beads (T cell Activation/Expansion Kit; Miltenyi Biotec) as previously described (20 (link)). T cells were stained with carboxyfluorescein succinimidyl ester at day 0. Proliferation was assessed at day 2 of co-culture by flow cytometry.
Brenig R., Pop O.T., Triantafyllou E., Geng A., Singanayagam A., Perez-Shibayama C., Besse L., Cupovic J., Künzler P., Boldanova T., Brand S., Semela D., Duong F.H., Weston C.J., Ludewig B., Heim M.H., Wendon J., Antoniades C.G, & Bernsmeier C. (2019). Expression of AXL receptor tyrosine kinase relates to monocyte dysfunction and severity of cirrhosis. Life Science Alliance, 3(1), e201900465.
Corresponding Organization : University of St. Gallen
Other organizations :
King's College Hospital, King's College London, St Mary's Hospital, Imperial College London, NIHR Birmingham Liver Biomedical Research Unit, National Institute for Health Research, University of Birmingham
AXL expression levels in monocytes (high: >20%/450 MFI, low: <7.5%/300 MFI)
dependent variables
Proliferation of T cells co-cultured with monocytes, assessed at day 2 of co-culture by flow cytometry
control variables
Monocytes from healthy controls (HCs) and patients with cirrhosis
CD3+ T cells from a different healthy donor
T cell stimulation induced with anti-CD2/CD3/CD28 beads
T cells stained with carboxyfluorescein succinimidyl ester at day 0
positive controls
Not explicitly mentioned
negative controls
Not explicitly mentioned
Annotations
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