Journal: bioRxiv

Article Title: Serial Thermal Ablation Induces Abscopal Antitumor Immunity and Reveals Targetable CSF1R-Dependent Resistance in Pancreatic Cancer

doi: 10.64898/2026.04.05.713683

Figure Lengend Snippet: A) Representative 20x NK1.1 immunohistochemistry staining images. Scale bars 100 µm. B) NK1.1 staining is significantly increased in 3 RFA contralateral tumors compared to 1 RFA contralateral tumors (***p<0.0001). NK1.1 is significantly increased in 3 RFA-treated tumors compared to 3 RFA contralateral tumors (*p=0.0141). n=10 fields analyzed per tumor. C) Representative 20x immunofluorescence images of CD4 (purple), CD8α (red), GZMB (green), DAPI nuclear stain (blue). D) CD4 + GZMB + cells are not increased in 3 RFA contralateral tumors. E) CD8α + GZMB + cells are significantly increased in the three RFA contralateral tumors compared to 1 RFA contralateral (****p<0.0001) and 3 RFA RFA-treated tumors (**p=0.0046). n=8 fields analyzed per group. F) Schematic of the protocol to evaluate the effects of CD8 depletion on serial RFA-treated and contralateral tumors. Created using BioRender . G) CD8α depletion significantly reduces the %CD8α + cells per live cells in RFA-treated tumors. H) α-CD8α significantly increases the tumor volume of RFA-treated (*p<0.05; ***p<0.001) and I) contralateral tumors after 2 RFA treatments but does not significantly increase the final tumor volumes after the third RFA treatment. Statistics were done using Prism GraphPad software.

Article Snippet: The following primary antibodies were used: CD4 (1:35, ab288724, abcam), CD8α (1:50, MAB116-100, R&D Systems), GZMB (1:100, AF1865, R&D Systems).

Techniques: Immunohistochemistry, Staining, Immunofluorescence, Software

Journal: bioRxiv

Article Title: Serial Thermal Ablation Induces Abscopal Antitumor Immunity and Reveals Targetable CSF1R-Dependent Resistance in Pancreatic Cancer

doi: 10.64898/2026.04.05.713683

Figure Lengend Snippet: A) Experimental design setup. B) Serial thermal ablation using RFA in combination with anti-PD-L1 and Quemli does not significantly reduce the volume of treated tumors. C) However, serial RFA in combination with anti-PD-L1 and Quemli significantly reduces the tumor volume of contralateral tumors compared to serial RFA + IgG+Vehicle (*p<0.05) and compared to serial RFA + Quemli treated alone (****p<0.0001). D) Representative 20x CSF1R immunohistochemistry images. E) Quantification of CSF1R + cells showed an increase in CSF1R + area per field in serial RFA tumors + anti-PD-L1 with or without Quemli compared to serial RFA + vehicle in both RFA-treated and F) contralateral tumors. A two-way Anova was used in Prism GraphPad for statistical comparisons. G) Experimental design setup. H) Serial thermal ablation using RFA in combination with CSF1R inhibition, anti-PD-L1 and Quemli significantly reduces the volume of ablated tumors (**p<0.01; ***p<0.001) and I) contralateral tumors (**p<0.01; ***p<0.001). J) Combination treatment using 3 RFA + anti-PD-L1, Quemli and CSF1R inhibition significantly increases the infiltration of CD8 + T cells in treated (**p<0.01; *p<0.05) and K) contralateral tumors (**p<0.01; *p<0.05) . L) Combination treatment using 3 RFA + anti-PD-L1, Quemli and CSF1R inhibition significantly increases GZMB staining in treated (***p<0.001; **p<0.01) and M) contralateral tumors (*p<0.05; **p<0.01; ***p<0.001). A two-way ANOVA in Prism GraphPad was used for statistical analysis. Scale bars 50 µm.

Article Snippet: The following primary antibodies were used: CD4 (1:35, ab288724, abcam), CD8α (1:50, MAB116-100, R&D Systems), GZMB (1:100, AF1865, R&D Systems).

Techniques: Immunohistochemistry, Inhibition, Staining