C57bl 6 foxp3 gfp

Nonobese diabetic, wt C57BL/6 (B6), C57BL/6-Foxp3-GFP, IFN-AR1^−/−, and Rag^−/− mice were purchased from Jackson Laboratories, and bred at the Johns Hopkins School of Medicine facility. All animal experiments were conducted in accordance with the National Institutes of Health guide for use and care of laboratory animals, and under a protocol approved by the JHU Animal Care and Use Committee.

Transgenic C57BL/6.Foxp3^GFP and wild type C57BL/6 mice were purchased from The Jackson Laboratory. Human CD2 transgenic C57BL/6.Foxp3^hCD2 [16 (link)] and 4-1BB deficient C57BL/6.4-1BB^-/- mice [17 (link)] were bred and cared for in University of Louisville’s specific-pathogen-free vivarium in accordance with NIH guidelines. All animal procedures were approved by the University of Louisville’s IACUC. Antibodies used include anti-CD3 agonistic Ab (clone 145-2C11, BD Pharmingen, Cat# 553058), anti-TGF-β (1,2,3) neutralizing Ab (clone 1D11, homemade), anti-IFN-γ blocking Ab (clone XMG 1.2, homemade), and anti-IL-2 neutralizing Ab (clone S4B6, BD Pharmingen, Cat# 554375). Fluorescently-labeled Abs to various cell surface markers were obtained commercially: α-CD4-APC (BD Pharmingen, 553051); α-CD25-PE (BD Pharmingen, 553866); α-CD25-PE-Cy7 (eBioscience, 25-0251-82); α-hCD2-PE (BioLegend, 300208). Where indicated Tconvs or Tregs were stained with 2.5μM carboxyfluorescein succinimidyl ester (CFSE) (Invitrogen, C1157). CTLL-2 cell line was obtained from ATCC (TIB-214). Human IL-2, SA-4-1BBL, and SA proteins were produced in our lab according to standard protocols as previously reported [10 (link),11 (link)].

MiceY145F knock‐in mice, SLP‐76^flox/Y145F conditional mutant (cY145F), and SLP‐76^flox/+ conditional heterozygous (cSLP76) mice were generated as previously described 8, 12 and bred in our facility. TCRβ/δ KO, I‐Abβ KO (MHC‐II KO), C57BL/6 CD90.1, C57BL/6 Foxp3.GFP and C57BL/6.SJL Foxp3.GFP reporter mice were purchased from The Jackson Laboratory (Bar Harbor, ME) or Charles River (Kingston, NY) and were bred and maintained in our animal facility. Mice were housed in pathogen‐free conditions and treated in strict compliance with Institutional Animal Care and Use Committee regulations of the University of Pennsylvania.