Anti cd3 apc clone 145 2c11

IFNγ^ΔKRKR mice were bred to TCR-I mice (B6.Cg-Tg(TcraY1,TcrbY1) 416Tev/J, 005236). TCR-I mice are transgenic for a TCR specific for epitope I of SV40 large T. For ATT experiments, female Rag1^–/– mice were injected subcutaneously with 1 × 10⁶ 16.113 cells. When tumors reached 500–600 mm³ in size, mice were treated with 1 × 10⁶ to 2 × 10⁶ TCR-I IFNγ or IFNγ^ΔKRKR T cells that were collected from TCR-I/IFNγ^ΔKRKR or TCR-I/IFNγ^WT mice. During ATT, weight and well-being were monitored from the day of transfer. Levels of serum IFNγ were determined using a mouse IFNγ ELISA (BD Biosciences). T cell expansion was monitored using anti-CD3-APC (clone 145-2C11, Biolegend), anti-CD8-BV421 (clone 53-6.7, Biolegend) and anti-Vβ7-PE (clone TR310, BD Biosciences).

In vivo labeling has been previously described (Cinamon et al., 2008 (link)). Briefly, mice were injected intravenously with 200 μL containing 2.5 μg of anti-CD19-PE (clone 6D5, Biolegend) and or anti-CD3-PE (clone 17A2, Biolegend) antibodies 5 min before culling the mice. Spleens were collected and stained on ice with anti-B220-Alexa700 (clone RA3-682, Biolegend), anti-CD3-APC (clone 145-2C11, Biolegend), anti-CD4-BV785 (clone RM4-5, Biolegend), anti-CD8-BV-605 (clone53-6.7, Biolegend). For stromal cells characterization of the Ccl19-iEYFP.2 mice, cells were stained with antibodies against CD45 (clone:30-F11, BioLegend), Ter119 (clone:TER-119, BioLegend), CD31 (clone:MEC13.3, BioLegend), PDPN (clone:8.1.1, BioLegend).Cells were acquired on a BD LSR Fortessa (BD Biosciences) and analyzed on FlowJo software (Tree Star Inc.).