Transgenic Mouse Models for T Cell Studies

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)].