be0356  (Bio X Cell)

Journal: Journal for immunotherapy of cancer

Article Title: Adenoviral-vectored neoantigen vaccine augments hyperexpanded CD8 + T cell control of tumor challenge in mice.

doi: 10.1136/jitc-2024-009644

Figure Lengend Snippet: Figure 1 B16-F10-OVA neoantigen vaccine trial: (A) Timeline of vaccination and challenge for prophylactic and therapeutic challenge. (B) C57BL/6 mice were immunized with 109 viral particles of Ad26.OVA or 50 μg of OVA protein with Poly I:C and anti- CD40 ab. Mice were bled 27 days later and whole blood intracellular cytokine staining was used to determine immunogenicity. OVA targeting cellular immune response was measured by IFNγ secreting CD8+ T cells isolated from whole blood. Individual mice are denoted by single dots. (C) C57BL/6 mice were challenged with 105 B16-F10-OVA cells implanted in the right flank. Mean tumor growth of B16-F10-OVA tumors. Bars display mean±the SEM. (D) Survival curve of B16-F10-OVA challenged mice. N=9 for sham. N=10 for OVA protein and Ad26.OVA respectively. (E) C57BL/6 mice were challenged with 105 B16-F10-OVA cells. Four days later the mice were immunized with 109 viral particles of Ad26.OVA, 50 μg of OVA protein with Poly I:C and anti- CD40 ab, or sham. (F) Survival curve of B16-F10-OVA challenged mice vaccinated therapeutically. Ad26, adenovirus serotype 26; ICS, intracellular cytokine staining; IFN, interferon; OVA, ovalbumin; Poly I:C, polyinosinic acid:polycytidylic acid; WB, whole blood.

Article Snippet: Mice received a single shot of 109 vp Ad26 replication- incompetent vaccine suspended in 1× phosphate- buffered saline (PBS), single shot peptide vaccine consisting of: 50 μg of each of the seven peptides in the 7Epi immunogen with 100 μg Poly(I:C) (InvivoGen: 31852- 29- 6) and 50 μg anti- CD40 antibody (Bio X Cell: BP0016- 2, clone FJK45), primeboost peptide vaccine with the same adjuvants, or sham vaccines.

Techniques: Staining, Immunopeptidomics, Isolation

Journal: Journal for immunotherapy of cancer

Article Title: Adenoviral-vectored neoantigen vaccine augments hyperexpanded CD8 + T cell control of tumor challenge in mice.

doi: 10.1136/jitc-2024-009644

Figure Lengend Snippet: Figure 3 MC38 vaccine protection: (A) Vaccination schedule for Ad26 and peptide vaccines. (B,C) C57BL/6 mice were immunized intramuscularly with 109 viral particles of Ad26 or 50 μg of each peptide with Poly I:C and anti-CD40 ab. Mice were bled 1 day prior to challenge and whole blood intracellular cytokine staining was used to determine immunogenicity. Whole blood intracellular cytokine staining (WB-ICS) showing prechallenge immunogenicity as measured by IFNγ+ and CD107a+ secreting CD8+ T cells after stimulation with 7Epi immunogen pool. Mann-Whitney test was performed *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001. (D) C57BL/6 mice were challenged with 3×105 MC38 tumor cells implanted in the right flank. After day 12 postchallenge, tumors were measured 2–3 times per week. Dots represent measurement time points. Bars display the mean±the SEM. Two-way ANOVA test used. Significance values represent significance compared with the sham group. Sham group in figures (D,E) consists of naïve, adjuvant, and Ad26.Luciferase immunized mice. Black * represent significance comparing Ad26.VP22.7Epi to sham and white * represent significance comparing peptide to sham. (E) Survival curve of MC38 challenged mice. Mantel-Cox test conducted to determine significance ****=p<0.0001. (F) Spearman correlation of mouse cytokines measured in (B,C) to tumor growth. Blue represents negative correlation and red represents positive correlation. FDR correction used. *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001. Tumor growth studies were the result of multiple independent experiments with the following total N: Sham=89, peptide+adjuvant=19, Ad26.7Epi=28, Ad26.7Epi.VP22=26, Ad26.VP22.7Epi=30, Ad26.Shark=28. WB-ICS was performed on a subset of mice. Ad26, adenovirus serotype 26; ANOVA, analysis of variance; FDR, false discovery rate; IFN, interferon; Poly I:C, polyinosinic acid:polycytidylic acid; Treg, regulatory T cell.

Article Snippet: Mice received a single shot of 109 vp Ad26 replication- incompetent vaccine suspended in 1× phosphate- buffered saline (PBS), single shot peptide vaccine consisting of: 50 μg of each of the seven peptides in the 7Epi immunogen with 100 μg Poly(I:C) (InvivoGen: 31852- 29- 6) and 50 μg anti- CD40 antibody (Bio X Cell: BP0016- 2, clone FJK45), primeboost peptide vaccine with the same adjuvants, or sham vaccines.

Techniques: Vaccines, Staining, Immunopeptidomics, MANN-WHITNEY, Adjuvant, Luciferase

Journal: Cell Reports Medicine

Article Title: TNFR1 signaling promotes pancreatic tumor growth by limiting dendritic cell number and function

doi: 10.1016/j.xcrm.2024.101696

Figure Lengend Snippet: Simultaneous blockade of TNFR1 and PD-1 reduced KPC tumor growth KPC mice in which established pancreatic tumors were detected by ultrasound were treated with either control ( n = 7) antibody or anti-TNFR1 plus anti-PD-1 antibody ( n = 11). (A–D) Tumors were harvested (ctrl, n = 4; anti-TNFR1, n = 3; and anti-TNFR1 plus anti-PD-1, n = 6) at day 24, and measured tumor volume (A), infiltrating total DCs (B, C), activated DCs (CD11c + CD40 + ) (D), and T cells (E) were analyzed. In the figure shown in C, pancreatic tumor tissues were stained for CD11c (green color) and DAPI (blue). Histogram showed the number of CD11c-positive cells per high power field (HPF). (F) Il12b expression of purified DCs measured by quantitative real-time PCR. Each data point represents pooled DCs from 3 mice. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. NS, not significant.

Article Snippet: InVivoPlus anti-mouse CD40 , BioXCell , Cat#BP0016-2 RRID: AB_1107601.

Techniques: Control, Staining, Expressing, Purification, Real-time Polymerase Chain Reaction

Journal: Cell Reports Medicine

Article Title: TNFR1 signaling promotes pancreatic tumor growth by limiting dendritic cell number and function

doi: 10.1016/j.xcrm.2024.101696

Figure Lengend Snippet: Comparative efficacy of TNFR1 blockade vs. Flt3L and agonistic anti-CD40 in treating subcutaneous KPC cell tumors (A–D) KPC cells were subcutaneously injected into WT or TNFR1-deficient mice and allowed to grow for 17 days. Schematic representation of therapeutic regimen is shown in (A), and the percent tumor growth since day 17 is shown in (B). On day 32, the percentage and number of tumor-infiltrating DCs (C) and their PD-L1 (D) and MHC-II expression (E) were measured. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. NS, not significant.

Article Snippet: InVivoPlus anti-mouse CD40 , BioXCell , Cat#BP0016-2 RRID: AB_1107601.

Techniques: Injection, Expressing

Journal: Cell Reports Medicine

Article Title: TNFR1 signaling promotes pancreatic tumor growth by limiting dendritic cell number and function

doi: 10.1016/j.xcrm.2024.101696

Figure Lengend Snippet:

Article Snippet: InVivoPlus anti-mouse CD40 , BioXCell , Cat#BP0016-2 RRID: AB_1107601.

Techniques: Purification, In Vivo, Functional Assay, Control, Recombinant, Staining, Flow Cytometry, Sequencing, Gene Expression, Generated, Knock-Out, Software

Journal: Nature Communications

Article Title: Senescent cancer-associated fibroblasts in pancreatic adenocarcinoma restrict CD8 + T cell activation and limit responsiveness to immunotherapy in mice

doi: 10.1038/s41467-024-50441-7

Figure Lengend Snippet: a Diagram of experimental design. 6422c1 KPC tumor cells were injected into mouse pancreata. Mice were treated with ABT-199, with ICT including αCTLA4, αPD1 and αCD40, or with both treatment types. b H&E-stained images of representative pancreatic tissues in mice treated with the different protocols. Dark regions represent remaining normal acinar tissue (white arrows). Tumor lesions in ABT + ICT mice are indicated by black arrows. Scale bar = 1 mm. c Pancreatic tissue weights upon mouse sacrifice in the different treatment mouse groups. Note that weights in ICT and ABT + ICT groups include some remaining normal tissue. n = 10 UNT, n = 6 ABT, n = 11 ICT, n = 12 ABT + ICT. d Higher magnification of H&E-stained tumor regions shown in ( b ). Dashed line indicates necrotic region in ABT + ICT-treated tumor. Scale bar = 100 μm. e Percentage of necrotic area out of tumor lesion area in the different treatment groups. Tissues that did not contain any remaining tumor lesions were not included. n = 10 UNT, n = 5 ABT, n = 9 ICT, n = 8 ABT + ICT. f Stain of tumor lesions for YFP in same, marking carcinoma cells with lesions. Scale bar = 100 μm. g Percentage of lesion area stained for YFP, indicating carcinoma cells, in the different treatment groups. n = 8 UNT, n = 6 ABT, n = 9 ICT, n = 8 ABT + ICT. All values indicate mean ± SEM. P values were calculated with Brown-Forsythe ANOVA test and Benjamini-Hochberg false discovery rate correction.

Article Snippet: Immunotherapy treatments to KPC-implanted mice followed previously-described protocols , , and included three antibodies: anti-PD-1 (clone RMP 1-14, BioXcell), administered 3 times a week for 2 weeks starting 10 days after implantation, at 200 μg per treatment; anti-CTLA4 (clone 9H10, BioXcell), 200 μg, administered 3 times within one week starting 10 days after implantation; anti-CD40 agonist antibody (clone FGK45, BioXcell), 100 μg, injected intraperitoneally once on day 13.

Techniques: Injection, Staining