Anti cd8β clone lyt 3

For anti PD-L1 therapy, anti-PD-L1 Ab (clone 10F.9G2, BioXcell) was given intraperitoneally (i.p.) every third day from the day RT performed at a dose of 200 μg/mouse. Rat IgG2b Ab (clone LTF-2, BioXcell) was used as a control. To deplete CD8⁺ and CD4⁺ T cells, 200 μg of anti-CD8β (clone Lyt 3.2, BioXcell) and anti-CD4 (clone GK1.5, BioXcell) Abs were administrated i.p. every third day for three times from the day when RT was given, respectively. To neutralize IL-12 and IFN-γ, 1 mg of anti-IL-12p40 (clone C17.8, BioXcell) and anti-IFN-γ (clone R4-6A2, BioXcell) Abs were injected by i.p. at the day when RT was performed, with follow-up doses of 500 μg for 5 consecutive days.

Mice were treated with either anti-LAP or IC mAbs prepared in PBS. Mouse anti-LAP monoclonal antibodies were isolated from hybridoma generated in-house. Two clones were employed for in vivo treatments: TW7-28G11 (IgG2b) and TW7-16B4 (IgG1). Respective ICs, MPC-11 (IgG2b) and MOPC-21 (IgG1), anti-CD103 (clone M290) and anti-PD1 (RMP-1) were purchased from BioXCell. As a standard treatment, antibodies were administered intraperitoneallly, 10 mg/kg every third day following tumor implantation. In some experiments, mice were treated i.p. with 100 μg per mouse of anti-CD8β (clone Lyt-3.2; BioXCell) Ab or IC (rat IgG1, HRPN; BioXCell) on days –1, 7, and 14 after B16F10 implantation.