Journal: Cancer Discovery
Article Title: PD-1 Blockade–Induced DKK1 Expression by CD8 + T Cells Promotes Blood–Brain Barrier Permeabilization
doi: 10.1158/2159-8290.CD-25-1222
Figure Lengend Snippet: Induction with anti–PD-1 antibody prior to cisplatin chemotherapy increases the intracranial therapeutic efficacy. A, A schematic illustration depicting various treatment groups and their therapeutic regimens, evaluating the efficacy of anti–PD-1 and cisplatin-based combination therapy sequences on experimental BrM of anti-PD-1–resistant LLC cells. B, An experimental BrM assay was performed on 8-week-old C57BL/6 mice treated with anti–PD-1 and cisplatin chemotherapy in opposing sequences. A Kaplan–Meier survival curve is plotted ( n = 6–7 mice/group). C–E, Eight-week-old C57BL/6 mice harboring subcutaneous LLC xenografts (∼100 mm 3 ) were treated with IgG, anti–PD-1, IgG + cisplatin, anti–PD-1 + cisplatin, and the opposing sequence, cisplatin + anti–PD-1, and the therapeutic response was monitored. Tumor growth across various treatment groups is plotted (Arrow indicates start of the treatment; n = 4–5 mice/group; C ). A bar graph showing the activation status of CD8 + T cells isolated from the spleens of these mice is plotted, as assessed by flow cytometry ( D ). A bar graph showing the percentage of dead cells [propidium iodide (PI)–positive] in a tumor cell killing assay, using CD8 + T cells isolated from the spleens of mice from the different conditions cocultured with LLC cells, as assessed by flow cytometry ( E ). F and G, Adoptive transfer of wild-type (Vector) and DKK1 KD [guide RNA (gRNA)] CD8 + T cells obtained from BALB/c mice was performed on 8-week-old SCID mice subjected to an experimental BrM assay using the 4T1 cancer cells. These mice were subsequently treated with a combination sequence of anti–PD-1 and cisplatin. An illustration depicting the experimental methodology for testing the effect of DKK1 KD on the therapeutic efficacy of the anti–PD-1 + cisplatin combination sequence is provided. Kaplan–Meier survival curve is plotted ( n = 6 mice/group; G ). H, Eight-week-old BALB/c mice, preconditioned with a single dose of IgG/anti–PD-1 or untreated (naïve), received cisplatin monotherapy 72 hours later. After 6 hours, the brains were harvested and prepared for the assessment of cisplatin concentration using LC/MS. A bar graph showing absolute quantification of cisplatin in the brains of BALB/c mice from various treatment groups is plotted ( n = 3–4 mice/group). Significance was assessed by means of the log-rank test for B and G , two-way ANOVA for C , and one-way ANOVA for D , E , and H . *, P < 0.05; **, P < 0.01; ***, P < 0.001. [ A, Created in BioRender. Raviv, Z. (2026) https://BioRender.com/rezdwrc ; F, Created in BioRender. Raviv, Z. (2026) https://BioRender.com/oo7uvma .]
Article Snippet: Murine EMT6 (cat. #CRL-2755; RRID:CVCL_1923), 4T1 (cat. #CRL-2539; RRID:CVCL_0125) mammary carcinoma, and LLC (cat. #CRL-1642; RRID:CVCL_4358) cell lines were purchased from the ATCC.
Techniques: Drug discovery, Sequencing, Clinical Proteomics, Activation Assay, Isolation, Flow Cytometry, Adoptive Transfer Assay, Plasmid Preparation, Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Quantitative Proteomics
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ATCC manufactures this product 
![Induction with anti–PD-1 antibody prior to cisplatin chemotherapy increases the intracranial therapeutic efficacy. A, A schematic illustration depicting various treatment groups and their therapeutic regimens, evaluating the efficacy of anti–PD-1 and cisplatin-based combination therapy sequences on experimental BrM of anti-PD-1–resistant <t>LLC</t> cells. B, An experimental BrM assay was performed on 8-week-old C57BL/6 mice treated with anti–PD-1 and cisplatin chemotherapy in opposing sequences. A Kaplan–Meier survival curve is plotted ( n = 6–7 mice/group). C–E, Eight-week-old C57BL/6 mice harboring subcutaneous LLC xenografts (∼100 mm 3 ) were treated with IgG, anti–PD-1, IgG + cisplatin, anti–PD-1 + cisplatin, and the opposing sequence, cisplatin + anti–PD-1, and the therapeutic response was monitored. Tumor growth across various treatment groups is plotted (Arrow indicates start of the treatment; n = 4–5 mice/group; C ). A bar graph showing the activation status of CD8 + T cells isolated from the spleens of these mice is plotted, as assessed by flow cytometry ( D ). A bar graph showing the percentage of dead cells [propidium iodide (PI)–positive] in a tumor cell killing assay, using CD8 + T cells isolated from the spleens of mice from the different conditions cocultured with LLC cells, as assessed by flow cytometry ( E ). F and G, Adoptive transfer of wild-type (Vector) and DKK1 KD [guide RNA (gRNA)] CD8 + T cells obtained from BALB/c mice was performed on 8-week-old SCID mice subjected to an experimental BrM assay using <t>the</t> <t>4T1</t> cancer cells. These mice were subsequently treated with a combination sequence of anti–PD-1 and cisplatin. An illustration depicting the experimental methodology for testing the effect of DKK1 KD on the therapeutic efficacy of the anti–PD-1 + cisplatin combination sequence is provided. Kaplan–Meier survival curve is plotted ( n = 6 mice/group; G ). H, Eight-week-old BALB/c mice, preconditioned with a single dose of IgG/anti–PD-1 or untreated (naïve), received cisplatin monotherapy 72 hours later. After 6 hours, the brains were harvested and prepared for the assessment of cisplatin concentration using LC/MS. A bar graph showing absolute quantification of cisplatin in the brains of BALB/c mice from various treatment groups is plotted ( n = 3–4 mice/group). Significance was assessed by means of the log-rank test for B and G , two-way ANOVA for C , and one-way ANOVA for D , E , and H . *, P < 0.05; **, P < 0.01; ***, P < 0.001. [ A, Created in BioRender. Raviv, Z. (2026) https://BioRender.com/rezdwrc ; F, Created in BioRender. Raviv, Z. (2026) https://BioRender.com/oo7uvma .]](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_3603/pmc13133603/pmc13133603__cd-25-1222_f7.jpg)