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Analysis of miR‐146a and miR‐142‐3p as Potential Markers of Freshly Isolated or In Vitro ‐Expanded Human Treg cells
Author(s) -
Holmstrøm K.,
Pedersen A. E.,
Gad M.
Publication year - 2017
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/sji.12517
Subject(s) - in vitro , foxp3 , microrna , biology , in vitro toxicology , immunology , microbiology and biotechnology , cancer research , immune system , gene , genetics
Regulatory CD4 + T cells (Tregs) are pivotal for prevention of autoimmunity. The use of Tregs is therefore of increasing interest in in vitro drug screening assays as well as for a cytotherapy per se against autoimmune disorders. For both purposes, in vitro expansion of peripheral blood Tregs is necessary and there is an increasing need to identify novel markers that can discriminate natural thymic‐derived Tregs ( tT regs) from other T cell subsets, and ideally, such markers should be stably expressed during in vitro expansion procedures. We screened for novel mi RNA s differentially expressed in tT regs and identified miR‐146a and 142‐3p as possible candidates. We analysed freshly isolated naïve and activated tT regs and non‐Treg subsets after or prior to in vitro expansion. We observed a tT reg‐specific profile of these mi RNA s together with FOXP 3 and Helios in freshly isolated tT regs, but observed a decline in the same markers in activated tT regs as opposed to naïve tT regs. In vitro ‐expanded Tregs could be identified based on FOXP 3 expression, but with loss of a discriminate profile for mi RNA candidates and a decline in FOXP 3 when activated tT regs were expanded. Our data demonstrate miR‐146a and 142‐3p as potential mi RNA markers for discrimination between non‐Treg cells and tT regs, but these mi RNA s are not stable markers for in vitro ‐expanded Treg cells. In addition, the loss of FOXP 3 in expansion of activated tT regs has implication for in vitro use of this cell subset in immunopharmacological assays and cytotherapy as FOXP 3 is pivotal for suppressive function.