rabbit mab against mouse cd3  (Cell Signaling Technology Inc)

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: Migratory DCs associate with tumor immune status, patient prognosis, and immunotherapeutic response in patients with cancer (A and B) Representative images of multiplex immunostaining of migratory DC and T cell markers on tumor sections derived from immune-“hot” and “cold” patients with PDAC/NSCLC are given. (C and D) Correlation analysis between migratory DC population and overall survival across our collected PDAC and NSCLC patient cohorts ( n = 66 patients with PDAC, n = 65 patients with NSCLC). (E and F) ROC curves represent the accuracy of migratory DCs and CD3 + CD8 + T cells in predicting overall survival in patients with PDAC and NSCLC. (G and H) Spatial analysis of multiplex IF images from each group. Magnified images are given. (I and J) Stack bar charts show the density of CD3 + CD8 + T cells at varying distances from migratory DCs within both hot and cold tumors from patients with PDAC/NSCLC. (K) Representative images of multiplex immunostaining of migratory DC and T cell markers on tumor sections from NSCLC immunotherapy non-responder and responder. (L) High population of migratory DCs correlated with better progression-free survival in patients with advanced NSCLC treated with anti-PD-1 ( n = 19 patients). (M) ROC curves illustrate the predictive accuracy of migratory DCs and CD3 + CD8 + T cells for progression-free survival in patients with advanced NSCLC treated with anti-PD-1. (N) Spatial proximity analysis of multiplex IF images from each group. Magnified image is given. (O) Stack bar chart illustrates the density of CD3 + CD8 + T cells at different distances from migratory DCs in each group. (C, D, and L) Log rank (Mantel-Cox). Scale bars in (A), (B), and (K) represent 50 μm. See also Figure S1 and Tables S1 , , and .

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Multiplex Assay, Immunostaining, Derivative Assay

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: cNP cancer cell @MV DC treatment triggers DC maturation and CD8 + T and NK cell activation in vitro (A and B) FACS analysis measured CD86 expression in immature human/mouse DCs after treatments. Bar charts display the percentage of CD11c + CD86 + DCs per group ( n = 3 independent experiments). (C and D) FACS analysis examined human/murine CD3 + CD8 + T cell proliferation after treatments ( n = 3 independent experiments). (E–H) FACS analysis assessed human/murine CD3 + CD8 + T cell activation after treatments. Bar charts show the percentage of CD3 + CD8 + CD69 + T cells or CD3 + CD8 + IFN-γ + T cells per group ( n = 3 independent experiments). (I–L) FACS analysis assessed human/murine NK cell activation after treatments. Graphs show the percentage of CD3 − CD56 + CD69 + , CD3 − NK1.1 + CD69 + , CD3 − CD56 + IFN-γ + , or CD3 − NK1.1 + IFN-γ + relative to the total human/murine NK population in each group ( n = 3 independent experiments). (M and N) Schematic diagrams illustrate LDH release assays conducted on PDAC patient-derived tumor organoids (T, target), using varying ratios of CD3 + CD8 + T cells (E, effector) pre-treated with the indicated treatments (M), or co-cultured with hDCs pre-treated with the indicated treatments (N). Bar charts indicate the killing efficiency percentage at each E:T ratio among different groups ( n = 3 independent experiments). (O and P) Cytotoxicity LDH release assays on DT6066 cells were conducted using different ratios of CD3 + CD8 + T cells that had been either pre-treated with the indicated treatments (O) or co-cultured with DCs pre-treated with the indicated treatments (P) ( n = 3 independent experiments). (A–L) One-way ANOVA. (M–P) Two-way ANOVA. See also Figure S3 .

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Activation Assay, In Vitro, Expressing, Derivative Assay, Cell Culture

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: cNP DT6066 @MV DC treatment converts immune-cold pancreatic tumors into hot tumors, reduces hypoxia, and enhances blood vessel function (A) FACS analysis of CD11c + CD86 + DCs in orthotopic DT6066 pancreatic tumors after treatments, with a bar chart showing their percentage relative to total lymphocytes ( n = 3 mice per group). (B) FACS analysis of CD3 + CD4 + CD69 + T cells in orthotopic pancreatic tumors after treatments, with a bar chart showing their percentage relative to total CD3 + CD4 + T cells in each group ( n = 3 mice per group). (C) FACS analysis of CD3 + CD8 + CD69 + T cells in orthotopic pancreatic tumors after treatments, with a bar chart showing their percentage relative to total CD3 + CD8 + T cells in each group ( n = 3 mice per group). (D) Representative images of CD11c and CD86 co-immunostaining in tumor sections from each treatment group. Bar chart shows the quantification of CD11c + CD86 + DC × 10 3 per cm 2 in each group ( n = 3 mice per group). (E and F) Representative images of co-immunostaining for CD3 and CD4 (E) or CD8 (F) in tumor sections from each treatment group ( n = 3 mice per group). (G) FACS analysis of CD3 − NK1.1 + NK cells or CD3 − NK1.1 + CD69 + NK cells in orthotopic pancreatic tumors after treatments. Bar charts show their percentage relative to total CD3 − cells (left) or CD3 − NK1.1 + NK cells (right) in each group ( n = 3 mice per group). (H–J) Cytotoxicity LDH release assays of DT6066 cells after co-culture with various ratios of CD3 + CD8 + T cells isolated from tumors (H), lymph nodes (I), or spleens (J) of orthotopic DT6066 pancreatic tumor-bearing mice after treatments ( n = 3 mice per group). (K) FACS analysis of migratory DCs in orthotopic pancreatic tumors and TDLNs from mice treated as indicated. Bar charts show CD86 + CD103 + DCs as a percentage of CD11c + DCs in each group ( n = 3 mice per group). (L) Representative images of CD11c/CD86/CD103 triple immunostaining on paired tumor and TDLN sections in each group. Bar chart shows the percentage of CD11c + CD86 + CD103 + migratory DC × 10 3 per cm 2 in tumors (left) or TDLNs (right) from each group ( n = 3 mice per group). (M–O) Representative microbubble contrast ultrasound images are shown, and bar charts show quantification across the entire tumors, including tumor cores ( n = 3 mice per group). (P) Representative IHC staining of endomucin in tumor sections from each treatment group. Bar chart shows blood vessel diameter (left) or number of blood vessels (right) per cm 2 in each group ( n = 3 mice per group). (Q) Representative image of GLUT1 and endomucin co-immunostaining in tumor sections from each group. Bar chart shows the relative GLUT1 intensity per group ( n = 3 mice per group). (A–G, K, L, and N–Q) One-way ANOVA. (H–J) Two-way ANOVA. Scale bars in (D), (E), (F), and (L) represent 50 μm. (M) 1 cm. (P and Q) 100 μm. See also Figures S5 and .

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Immunostaining, Co-Culture Assay, Isolation, Triple Immunostaining, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet:

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Virus, Recombinant, Negative Staining, Lysis, Control, Phospho-proteomics, Cell Isolation, DNA Purification, DNA Extraction, Bicinchoninic Acid Protein Assay, Protein Purification, Magnetic Beads, Software