Journal: The Journal of Clinical Investigation

Article Title: Depleting tumor-specific Tregs at a single site eradicates disseminated tumors

doi: 10.1172/JCI64859

Figure Lengend Snippet: (A) Mice were challenged s.c. with 5 × 106 A20 tumor cells on the right and left flanks. Therapy was started when tumors reached 0.5–0.7 cm in diameter (usually between day 5 and 8). Treated mice received CpG i.t. only in their right tumor for 5 consecutive days. On day 1 and 5 of CpG therapy, αOX40 and αCTLA4 mAbs were either injected i.p. or i.t. (together with CpG in the right tumor). The systemic antitumor immune response generated by these systemic (i.p.) and local (i.t.) maneuvers was assessed by measuring the size of the contralateral (noninjected) left tumor and mouse disease-free survival. Results were pooled from 2 distinct experiments (n = 10 mice per group). (B) Tumor growth of the distant tumors (nontreated left tumors) when mAbs were injected systemically (i.p.) or locally (i.t. into the right tumor). Black arrows indicate day 1 of therapy. Both strategies (mAbs injected i.p. or i.t.) result in disappearance of the distant (left) tumors. (C) Relapse-free survival of mice treated with either local or systemic immunomodulation. Most of the mice treated systemically (i.p.) with αOX40/CTLA4 relapsed in the left tumor-draining lymph nodes, whereas almost all the mice who received αOX40 and αCTLA4 locally (i.t.) had a long-term survival (*P = 0.002). The number of mice per group is shown into parenthesis. (D) Therapeutic efficacy of 1:100 and 1:1,000 doses of αOX40 and αCTLA4 either injected i.p. or i.t. together with CpG and the resulting long-term disease-free survival. ttt, treatment.

Article Snippet: Anti-FOXP3 (clone FJK-16s, eBiosciences) intracellular stainings were performed as recommended by the mAb manufacturer. αOX40 (clone OX86, eBioscience) and αCTLA4 (clone UC10-4F10-11, BD Biosciences) were used for surface staining (together with intracellular anti-FOXP3).

Techniques: Injection, Generated

Journal: The Journal of Clinical Investigation

Article Title: Depleting tumor-specific Tregs at a single site eradicates disseminated tumors

doi: 10.1172/JCI64859

Figure Lengend Snippet: Mice were treated as in Figure ​Figure3A,3A, and. i.t. injections of therapy were done in right (local) tumors (red), and systemic antitumor effect was assessed by measuring growth of left (distant) tumors (blue). CpG was injected at 100 μg daily for 5 consecutive days. Low doses of mAbs (4 μg αOX40 or rat isotype or/and 1 μg αCTLA4 or hamster isotype) were injected on day 1 and 5 of CpG therapy into the same tumor. (A) Growth of distant tumors without therapy. (B–F) Systemic antitumor effect of CpG injections (B) alone on injected and distant tumors; (C) in combination with rat and hamster isotypes of αOX40 and αCTLA4 mAbs, respectively; (D) in combination with αOX40; (E) in combination with αCTLA4; and (F) in combination with αOX40 and αCTLA4. Previous curves pooled from at least 2 different experiments per group. (A–F) The number of surviving mice at day 60 is shown in parenthesis. (G) Survival of mice bearing 2 s.c. tumors (right and left flanks) that received CpG plus rat/hamster isotypes, CpG plus αOX40, CpG plus αCTLA4, or CpG plus αOX40/CTLA4 in right tumors. Survival with CpG plus αOX40/CTLA4 was significantly higher than with CpG plus αOX40 (P = 0.004) or CpG plus αCTLA4 (P = 0.03). Data are pooled from at least 2 different experiments per group; the number of mice per group is shown into parenthesis (*P < 0.05). Systemic antitumor effect of (H) αOX40 plus αCTLA4 local low-dose therapy without CpG (n = 4) and (I) s.c. CpG and i.t. αOX40 plus αCTLA4. (J) Survival of tumor-bearing mice treated with i.t. CpG and low-dose αOX40 plus αCTLA4 in the context of CD4 or CD8 T cell depletion (*P < 0.05).

Article Snippet: Anti-FOXP3 (clone FJK-16s, eBiosciences) intracellular stainings were performed as recommended by the mAb manufacturer. αOX40 (clone OX86, eBioscience) and αCTLA4 (clone UC10-4F10-11, BD Biosciences) were used for surface staining (together with intracellular anti-FOXP3).

Techniques: Injection

Journal: Biodrugs

Article Title: Targeting Co-Stimulatory Receptors of the TNF Superfamily for Cancer Immunotherapy

doi: 10.1007/s40259-022-00573-3

Figure Lengend Snippet: Tumor necrosis factor super family ( TNF-SF) agonistic monospecific antibodies in clinical studies ( www.clinicaltrials.gov )

Article Snippet: , , Adv. or metastatic malignancies , Ipilimumab (αCTLA-4)/Nivolumab (αPD-1) , I/II , Completed (11/2021) , NCT03126110 , Incyte Corporation.

Techniques: Clinical Proteomics

Journal: Biodrugs

Article Title: Targeting Co-Stimulatory Receptors of the TNF Superfamily for Cancer Immunotherapy

doi: 10.1007/s40259-022-00573-3

Figure Lengend Snippet: Tumor necrosis factor super family ( TNF-SF) agonistic monospecific antibodies in clinical studies ( www.clinicaltrials.gov )

Article Snippet: , , Adv. solid tumors , Durvalumab (αPD-1)/Tremelimumab (αCTLA-4) , I , Completed (08/2019) , NCT02705482 , MedImmune LLC.

Techniques: Clinical Proteomics

Journal: Biodrugs

Article Title: Targeting Co-Stimulatory Receptors of the TNF Superfamily for Cancer Immunotherapy

doi: 10.1007/s40259-022-00573-3

Figure Lengend Snippet: Agonistic tumor necrosis factor super family (TNF-SF) ligand-fusion proteins in clinical studies ( www.clinicaltrials.gov )

Article Snippet: , , Adv. solid tumors , Durvalumab (αPD-1)/Tremelimumab (αCTLA-4) , I , Completed (08/2019) , NCT02705482 , MedImmune LLC.

Techniques:

Journal: Biodrugs

Article Title: Targeting Co-Stimulatory Receptors of the TNF Superfamily for Cancer Immunotherapy

doi: 10.1007/s40259-022-00573-3

Figure Lengend Snippet: Tumor necrosis factor super family ( TNF-SF) agonistic monospecific antibodies in clinical studies ( www.clinicaltrials.gov )

Article Snippet: , , Adv. malignancies , Nivolumab (αPD-1)/Ipilimumab (αCTLA-4) , I/II , Completed (09/2019) , NCT03241173 , Incyte Corporation.

Techniques: Clinical Proteomics

Journal: PLoS Pathogens

Article Title: Synergistic Reversal of Intrahepatic HCV-Specific CD8 T Cell Exhaustion by Combined PD-1/CTLA-4 Blockade

doi: 10.1371/journal.ppat.1000313

Figure Lengend Snippet: Flow cytometry plots showing HCV 1073-specific CD8 T cell phenotype directly ex vivo and antigen-specific functions following 7 days of antigenic stimulation in the presence of isotype or blocking antibodies, using liver-derived (A) and blood-derived (B) lymphocytes from chronic patient C57. (Top panels): frequency of HCV 1073-specific CD8 T cells determined by cognate HLA-A2 tetramer staining. (Middle panels) far left: PD-1 and CTLA-4 expression ex vivo in gated tetramer + CD8 T cells (dot plots). Remaining right panels: HCV-specific IFN-γ production and CD107a mobilization in gated tetramer + CD8 T cells on day 7. (Bottom panels): Perforin expression in tetramer + (blue line) and total CD8 T cells (gray shaded) on day 7. (C) Fold increase in the expansion and effector functions of liver-derived (left) and blood-derived (right) HCV-specific CD8 T cells by αPD-L1 alone (white bar), αCTLA-4 alone (gray bar) and combined αPD-L1/αCTLA-4 blockade (black bar) relative to the isotype control for 3 chronic patients. The frequencies of functional tetramer + CD8 T cells in each culture were calculated by multiplying %tetramer + CD8 T cells with %IFN-γ + /tetramer + CD8 T cells, %perforin + /tetramer + CD8 T cells or %CD107a + /tetramer + CD8 T cells. (D) Flow cytometry plots showing CMV-specific CD8 T cells directly ex vivo and their antigen-specific functions following 7 days in vitro cultures from chronic patient C99.

Article Snippet: Of note, PD-1 and CTLA-4 expression in all subjects was examined using FITC-labeled αPD-1 (clone M1H4, BD) and PE-labeled αCTLA-4 (αCD152; clone BNI3, BD).

Techniques: Flow Cytometry, Ex Vivo, Blocking Assay, Derivative Assay, Staining, Expressing, Functional Assay, In Vitro