Supplementary Figure 2 from Curing Mice with Large Tumors by Locally Delivering Combinations of Immunomodulatory Antibodies

Expression of surface molecules in TLN and spleen of mice receiving the mAb treatments.</p

Supplementary Figure Legend from Curing Mice with Large Tumors by Locally Delivering Combinations of Immunomodulatory Antibodies

Figure legends for four supplementary figures.</p

Supplementary Figure 3 from Curing Mice with Large Tumors by Locally Delivering Combinations of Immunomodulatory Antibodies

Depleted of CD19 cells by the 4 mAb combination.</p

Supplementary Figure 4 from Curing Mice with Large Tumors by Locally Delivering Combinations of Immunomodulatory Antibodies

Expression of surface molecules in TIL of mice receiving mAb combinations.</p

Supplementary Figure 1 from Curing Mice with Large Tumors by Locally Delivering Combinations of Immunomodulatory Antibodies

Lymphocyte components of mice transplanted with tumor cells.</p

Local Release of Highly Loaded Antibodies from Functionalized Nanoporous Support for Cancer Immunotherapy

Local Release of Highly Loaded Antibodies from Functionalized Nanoporous Support for Cancer Immunotherap

Antitumor effects of anti-CD137/PD-1 mAbs againstne ID8 ovarian cancer.

<p>Mice (5/group) transplanted i.p. with 3 × 10⁶ ID8 cells 10 days previously were injected i.p. twice at 4 days interval with the indicated mAb combinations (0.5 mg of each mAb/mouse); survival was recorded (A, C) and mean survival time was calculated (B, D). The experiment was repeated once with similar results. E, Mice (8-9/group) transplanted i.p. with 3 × 10⁶ ID8 cells 3 days previously were injected i.p. twice at 4 days interval with 0.5 mg of control, anti-PD-1, anti-CD137 and anti-PD-1/CD137 mAb and their survival was recorded. *P < 0.05, **P < 0.01, compared with control mAb treated mice. </p

Combining anti-PD-1/CD137 mAb with cisplatin induced complete remission of established ID8 cancer with long-lasting systemic tumor-specific immunity.

<p>Mice (10/group) transplanted i.p. with 3 x 10⁶ ID8 cells 10 days earlier were injected i.p. with two doses of control, anti-PD-1, anti-CD137 or anti-PD-1/CD137 mAb (0.5 mg per dose per mouse) at 4 days interval with or without coadministration of cisplatin (10mg/kg) at the first treatment and their survival was evaluated (A). Mice (10/group) treated with combined anti-PD-1/CD137/cisplatin were depleted of lymphocyte subsets by injection of anti-CD4 (0.2 mg/mouse), anti-CD8 (0.2 mg/mouse), anti-NK1.1 (0.1 mg/mouse) or control mAb (0.2 mg/mouse) 48 and 72 hours prior to the first treatment and every 3-4 days thereafter for the duration of the experiments. Untreated tumor-bearing mice were as negative controls (UNT group). The survival of mice was recorded (B). Fifteen long-term surviving mice (120 days after original transplantation of ID8 cells) pooled from 2 experiments were challenged (5 mice/group) with ID8 cells given i.p. or s.c. or with TC1 cells transplanted s.c. (C); naive mice were transplanted with tumor cells as controls (D) and their survival was recorded. Mice (6/group) with established TC1 tumors of 4-5 mm mean diameter were injected i.p. with two doses of control, anti-PD-1, anti-CD137 or anti-PD-1/CD137 mAb (0.5 mg per dose per mouse) at 4 days interval with or without coadministraation of cisplatin (10 mg/kg) at the first treatment; tumor growth was measured (E) and survival was recorded (F). Data are representative of 2 experiments for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084927#pone-0084927-g006" target="_blank">Figure 6A-D</a>.</p

Combinatorial PD-1 Blockade and CD137 Activation Has Therapeutic Efficacy in Murine Cancer Models and Synergizes with Cisplatin

<div><p>There is an urgent need for improved therapy for advanced ovarian carcinoma, which may be met by administering immune-modulatory monoclonal antibodies (mAbs) to generate a tumor-destructive immune response. Using the ID8 mouse ovarian cancer model, we investigated the therapeutic efficacy of various mAb combinations in mice with intraperitoneal (i.p.) tumor established by transplanting 3 × 10⁶ ID8 cells 10 days previously. While most of the tested mAbs were ineffective when given individually or together, the data confirm our previous finding that 2 i.p. injections of a combination of anti-CD137 with anti-PD-1 mAbs doubles overall survival. Mice treated with this mAb combination have a significantly increased frequency and total number of CD8⁺ T cells both in the peritoneal lavage and spleens, and these cells are functional as demonstrated by antigen-specific cytolytic activity and IFN-γ production. While administration of anti-CD137 mAb as a single agent similarly increases CD8⁺ T cells, these have no functional activity, which may be attributed to up-regulation of co-inhibitory PD-1 and TIM-3 molecules induced by CD137. Addition of the anti-cancer drug cisplatin to the 2 mAb combination increased overall survival >90 days (and was probably curative) by a mechanism which included a systemic CD8⁺ T cell response with tumor specificity and immunological memory. Strikingly, combined treatment of cisplatin and CD137/PD-1 mAb also gave rise to the long-term survival of mice with established TC1 lung tumors. A similar combination of the 2 mAbs and cisplatin should be considered for clinical ‘translation’. </p> </div

Analysis of peritoneal lymphocyte subsets from mice injected with the anti-PD-1/CD137 combination.

<p>Mice (3/group) transplanted i.p. with 3 × 10⁶ ID8 cells 10 day earlier were injected i.p. twice at 4 days interval with 0.5 mg of control, anti-CD137, anti-PD-1 or anti-PD-1/CD137 mAb. Two weeks later, peritoneal lavage from treated mice was analyzed by flow cytometry for the percentage and phenotype of peritoneal lymphocytes. The percentages of CD3⁺, CD4⁺, CD8⁺ and CD19⁺ lymphocytes in peritoneal lavage and CD44⁺CD62L^- effector/memory cells in CD4⁺ and CD8⁺ T cells are shown in (A) and (B) respectively. The percentage of PD-1⁺TIM-3⁺ and PD-1^-TIM-3⁺ cells in peritoneal CD4⁺ and CD8⁺ T cells are shown in (C) with representative dotplots in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084927#pone.0084927.s002" target="_blank">Figure S2</a>. Data are presented as M±SEM from 3 mice of each group and are representative of 2 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, PD-1 or CD137 mAb compared with control mAb, PD-1/CD137 mAb compared with control and single mAb. </p