Transcriptomic analysis of Treg and Tconv cells from inflamed tissues of mice that are deficient for TNFR2 in Treg cells

Methods: Foxp3Cre-ERT2 x Tnfrsf1bFlox/Flox (icKO) and Foxp3Cre-ERT2 control (Ctrl) mice were immunized with MOG35-55 peptide to induce active EAE. From day 7 to 14, they were treated with Tamoxifen to delete TNFR2 only in Tregs. At day 14, Tregs and Tconvs were purified by flow cytometry from the inflamed central nervous system (CNS) or draining lymph nodes (dLN) for RNA extraction Results: In the CNS, TNFR2-deficient Tregs have lower expression of several Treg cell-signature genes and increased expression of genes normally expressed by Tconvs. The expression of 185 and 229 genes was respectively down- and up-regulated in Tconvs isolated from the CNS of mutant animals, compared to controls. Tconvs from icKO mice displayed a highly activated phenotype. Overall design: RNA-Seq of total mRNA of Tregs and Tconvs from the CNS or dLN at day 14 of EAE from mice that have TNFR2-deficient Tregs or control mice