Journal: JCI Insight

Article Title: Differentiated agonistic antibody targeting CD137 eradicates large tumors without hepatotoxicity

doi: 10.1172/jci.insight.133647

Figure Lengend Snippet: (A) Overall survival and tumor growth rates of individual mice with large (~500 mm3) CT-26 tumors following i.p. administration of 150 μg/mouse CTX-471, CTX-471-AF, and CTX-471-AF2 on days 0, 3, 6, and 9. (B) Tumor growth rates of individual mice with large (~450 mm3) CT-26 tumors following i.p. administration of CD137 agonists (CTX-471-AF or 3H3; 25 μg/mouse on days 0, 7, and 14), checkpoint inhibitors (Avelumab, RMP1-14, or 9H10; 200 μg/mouse on days 0, 3, and 6), or an OX40 agonist (OX-86; 200 μg/mouse on days 0, 3, and 6). (C) Kaplan-Meier survival curves of mice in B. Statistical significance was determined using Log-rank test (A and C) followed by Bonferroni’s multiple comparison compared with control treatment groups (*P<0.05, **P<0.01). (D and E) Histological analysis of CT-26 tumors on days 7, 10, and 14 following i.p. administration of 25 μg/mouse CTX-471-AF or isotype control on day 0. Formalin-fixed paraffin-embedded tissues were stained for H&E (D) or with antibodies against CD8 (E). Scale bars: 200 μm.

Article Snippet: For the comparative large-tumor experiment, clinical grade Avelumab (anti–PD-L1) was sourced from Myoderm (NDC 44087-3535-01), whereas RMP1-14 (anti–PD-1, catalog BE0146), 9H10 (anti–CTLA-4, catalog BP0131), and OX-86 (anti–OX-40, catalog BE0031) were purchased from BioXcell.

Techniques: Formalin-fixed Paraffin-Embedded, Staining

Journal: Frontiers in Immunology

Article Title: Engineered antibody cytokine chimera synergizes with DNA-launched nanoparticle vaccines to potentiate melanoma suppression in vivo

doi: 10.3389/fimmu.2023.1072810

Figure Lengend Snippet: Transcriptomic analyses demonstrated upregulation of Type I interferon responses following TA99-HL2-KOA1 treatment. (A) Map of ISG15 as part of the Type I interferon responsive elements in pathway analysis. (B, C) Fold changes in ISG15 in intratumoral DCs, macrophages and monocytes in mice treated with anti-PD1+ Trp2Vax + TA99-HL2-KOA1 versus corresponding cell types from mice treated with only anti-PD1 (B) or with anti-PD1+ Trp2Vax + TA99-WT (C) . (D, E) Fold changes of the most upregulated genes in the intratumoral macrophages of mice treated with anti-PD1+ Trp2Vax + TA99-HL2-KOA1 versus intratumoral macrophages from mice treated with only anti-PD1 (D) or with anti-PD1+ Trp2Vax + TA99-WT (E) . Asterisk * marks IFN-responsive genes. (F) Re-clustering of cells as in Figure 3E based on canonical macrophage markers. (G) Nos2 and Arg1 expression clustering based on (F) . (H) Percent M1 macrophages in each treatment group relative to the total of M1 and M2 macrophages. N=5 mice/group were pooled for transcriptomic analyses in this Figure. *p=0.021, ****p=10 -7 (PD-1 vs. TA99 Regimen), ****p<10 -10 (TA99 Regimen vs. HL2-KOA1 Regimen) by Fisher Exact Test. Each dot represents a pool of 5 animals receiving the same treatment.

Article Snippet: For recombinant anti–PD-1 (RMP1-14, Bio X Cell, Catalog: BE0146) administration, 200 µg of antibody was injected intraperitoneally formulated in 100 µL PBS to each mouse weekly beginning 7 days post tumor inoculation.

Techniques: Expressing

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: Stimulating Antitumoral Immunity by Percutaneous Cryoablation and Combination Immunoadjuvant Therapy in a Murine Model of Hepatocellular Carcinoma

doi: 10.1016/j.jvir.2023.05.008

Figure Lengend Snippet: Flow diagram of subject exclusion for time-to-endpoint analysis. *A cryoablation + programmed cell death 1 (n = 1) mouse died before the 6-week time point and was included in the time-to-endpoint data (n = 11 subjects) but not the relative tumor volume data (n = 10 subjects). Cryo = cryoablation; NASH = nonalcoholic steatohepatitis; PD-1 = programmed cell death 1.

Article Snippet: For PD-1 inhibitor treatment, 10 mg/kg of anti-PD-1 antibody (BE0146, InVivoMAb anti-mouse PD-1; Bio X Cell, Deer Park, Illinois) was injected percutaneously into the spleen or portal vein to maximize hepatic uptake.

Techniques:

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: Stimulating Antitumoral Immunity by Percutaneous Cryoablation and Combination Immunoadjuvant Therapy in a Murine Model of Hepatocellular Carcinoma

doi: 10.1016/j.jvir.2023.05.008

Figure Lengend Snippet: Relative tumor volume change at 1 week after treatment. (a) Nonablated satellite and (b) ablated tumor volumes from untreated mice and mice that underwent cryoablation (cryo), cryo + intratumoral CpG injection, cryo + programmed cell death 1 (PD-1) injection, or cryo + CpG + PD-1 treatment were calculated at 1 week after treatment, and fold change in the pretreatment tumor volume was determined. Treatment with cryo + CpG was associated with a statistically significant tumor growth retarding effect in both ablated and nonablated tumors compared with cryo alone (*P < .05, **P < .01). (c) Single tumor treatment with cryo CpG or cryo + CpG + PD-1 delayed the time to endpoint, with log-rank hazard ratios of 0.42 (P = .031) and 0.27 (P < .001), respectively, compared with cryo (C).

Article Snippet: For PD-1 inhibitor treatment, 10 mg/kg of anti-PD-1 antibody (BE0146, InVivoMAb anti-mouse PD-1; Bio X Cell, Deer Park, Illinois) was injected percutaneously into the spleen or portal vein to maximize hepatic uptake.

Techniques: Injection

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: Stimulating Antitumoral Immunity by Percutaneous Cryoablation and Combination Immunoadjuvant Therapy in a Murine Model of Hepatocellular Carcinoma

doi: 10.1016/j.jvir.2023.05.008

Figure Lengend Snippet: Histology. (a) Hematoxylin and eosin staining of an incompletely ablated intrahepatic tumor from a mouse treated with cryoablation (cryo) + CpG + programmed cell death 1 (PD-1). At 2× (left) and 8× (right) magnifications of normal hepatic parenchyma, tumor tissue characterized by small cells with large nuclei and a large nuclear-to-cytoplasmic ratio, and the highly eosinophilic and acellular ablation zone. A few small infiltrating cells are seen, many of which are lymphocytes. The liver-tumor interface (arrows), ablation-tumor (black arrowheads), and ablation-liver (gray arrowheads) interfaces are well demarcated at 8× magnification. (b) Qualitatively, cytotoxic CD8+ T cells accumulated at the tumor periphery in CpG-treated mice at 1 week after treatment. The liver-tumor interface is delineated by arrows in the hematoxylin and eosin staining image. Arrowheads show cytotoxic CD8+ T cells accumulated at the liver-tumor interface of the cryo + CpG + PD-1–treated subject. (c) Immunohistochemistry directed at CD8 and FOXP3 of normal liver and pooled viable ablated and nonablated tumor tissue from animals sacrificed at 1 week after treatment. Positive pixels were detected by software and normalized to the area of analyzed tissue to create positive pixel density (*P < .05). A = ablation zone; HE = hematoxylin and eosin; L = normal hepatic parenchyma; T = tumor tissue.

Article Snippet: For PD-1 inhibitor treatment, 10 mg/kg of anti-PD-1 antibody (BE0146, InVivoMAb anti-mouse PD-1; Bio X Cell, Deer Park, Illinois) was injected percutaneously into the spleen or portal vein to maximize hepatic uptake.

Techniques: Staining, Immunohistochemistry, Software

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: Stimulating Antitumoral Immunity by Percutaneous Cryoablation and Combination Immunoadjuvant Therapy in a Murine Model of Hepatocellular Carcinoma

doi: 10.1016/j.jvir.2023.05.008

Figure Lengend Snippet: Multiparametric flow cytometric analysis of ablated and nonablated (satellite) tumor tissue collected at 1 week after treatment. CD4+ and CD8+ were expressed as the percentage of CD3+ cells, NK+ CD137+ as the percentage of NK+ cells, and CD44+ PD-1+ as the percentage of CD8+ cells. CpG treatment was associated with cytotoxic CD8+ T-cell enrichment in ablated tumor tissue. CpG injection was strongly associated with increased programmed cell death 1 expression by activated cytotoxic T cells (CD44+ CD8+) in all tested tissue types (*P < .05, **P < .01, ***P < .001). Cryo = cryoablation; PD-1 = programmed cell death 1.

Article Snippet: For PD-1 inhibitor treatment, 10 mg/kg of anti-PD-1 antibody (BE0146, InVivoMAb anti-mouse PD-1; Bio X Cell, Deer Park, Illinois) was injected percutaneously into the spleen or portal vein to maximize hepatic uptake.

Techniques: Injection, Expressing

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: Stimulating Antitumoral Immunity by Percutaneous Cryoablation and Combination Immunoadjuvant Therapy in a Murine Model of Hepatocellular Carcinoma

doi: 10.1016/j.jvir.2023.05.008

Figure Lengend Snippet: Proinflammatory and anti-inflammatory serum cytokines at 1 week after treatment. (a) The serum levels of the proinflammatory cytokine interferon-γ and anti-inflammatory cytokines tumor growth factor-β1 and interleukin-10 were quantitated at baseline, 24 hours, 48 hours, and 1 week following treatment. (b) A linear correlation model (Pearson) was applied to cytokine concentrations at baseline; 24, 48, and 72 hours; and 1 and 2 weeks after treatment and compared with overall survival (top) and fold change of ablated tumor volume (bottom) at 1 week compared with that at the baseline. Statistically significant (P < .05) results are shown. Cryo = cryoablation; IFN-γ = interferon-γ; PD-1 = programmed cell death 1; TGF-β1 = transforming growth factor-β1.

Article Snippet: For PD-1 inhibitor treatment, 10 mg/kg of anti-PD-1 antibody (BE0146, InVivoMAb anti-mouse PD-1; Bio X Cell, Deer Park, Illinois) was injected percutaneously into the spleen or portal vein to maximize hepatic uptake.

Techniques:

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: Stimulating Antitumoral Immunity by Percutaneous Cryoablation and Combination Immunoadjuvant Therapy in a Murine Model of Hepatocellular Carcinoma

doi: 10.1016/j.jvir.2023.05.008

Figure Lengend Snippet: Proposed mechanism for cytotoxic T-cell (CTL) antitumoral immunity. (a) Tumors were cryoablated and coinjected with CpG, thereby releasing tumor antigens and stimulating proinflammatory cytokines (eg, interferon-γ). (b) Tumor antigens are taken up by circulating plasmacytoid dendritic cells, which are stimulated by CpG and proinflammatory cytokines to crosspresent the tumor antigens and activate CTLs. (c) Programmed cell death 1 inhibition at the time of tumor antigen cross-presentation prevents programmed death ligand 1–mediated CTL anergy and clonal deletion. (d) Tumor antigen crosspresentation and activation leads to CTL-mediated antitumoral immunity. APC = antigen-presenting cell; IFN-γ = interferon-γ; Inh. = inhibitor; PD-1 = programmed cell death 1; PD-L1 = programmed death ligand 1

Article Snippet: For PD-1 inhibitor treatment, 10 mg/kg of anti-PD-1 antibody (BE0146, InVivoMAb anti-mouse PD-1; Bio X Cell, Deer Park, Illinois) was injected percutaneously into the spleen or portal vein to maximize hepatic uptake.

Techniques: Inhibition, Activation Assay