Checkpoint blockade combined with adoptively transferred autologous PBMC or TALs slowed the OVC growth in NSG mice. (A) Experimental schema for tumor implantation, adoptive cell transfer (ACT) and antibody blockade treatment. tumor cells were IP or SQ injected into NSG mice. Twenty-one (or 14 in E) days after tumor implantation, mice were treated (IP) with autologous PBMCs or TALs and IL2 (2000 U). IgG or ICB antibodies (anti-PD1, 200 µg/mouse; anti-CTLA4, 100 µg/mouse), and IL2 were IP starting day 16 or 23 every 2–3 days for seven times. Thereafter mice were treated with IL2 once per week. Tumor volume was assessed by bioluminescent imaging (P/S, photons/s in B, C and E) or caliper measurement (D) of tumor-bearing mice at indicated time points post implantation. (B) Adoptive transfer of IL2-expanded autologous PBMC (1×106) in combination with anti-PD-1 blockade mildly delayed the growth of 099-luc-CL (IP) in NSG mice (n=3–4). (C) Adoptive transfer of IL2-expanded autologous PBMC (1×106), in combination with anti-PD-1/CTLA4 blockade significantly delayed the growth of 099-luc-CL (IP) in NSG mice (n=3). (D) Adoptive transfer of autologous PBMC (1×106) in combination with anti-PD-1/CTLA4 blockade significantly delay the progression of 362-PDX0-CL (SQ) in NSG mice (n=2–3). Tumor volume data were determined according to formula: (length × wide2)/2. (E) Adoptive transfer of IL2-expanded autologous TALs in combination with anti-PD-1/CTLA4 dual blockade significantly delayed the growth of 099-CL-Luc (SQ) in N-HSGM3 mice (n=3). Autologous TALs (4×105) were IP-injected at day 14 and IgG or CPB starting at day 16 for seven times. Tumor volume was assessed by bioluminescent imaging (protons/sec) since the tumor express luciferase at indicated time points post implantation. Tumor volume data were from mice reach end points and are presented as mean±SEM from one of the duplicated experiments. *P<0.05, **p<0.01, ns, not significant, using multiple t-test (B) or two-way ANOVA (C, D, E). ANOVA, analysis of variance; IL, interleukin; IP, intraperitoneally; OVC, ovarian cancer; PBMC, peripheral blood mononuclear cell; SQ, subcutaneously; TALs, tumor-associated leukocytes; TILs, tumor-infiltrating leukocytes.