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

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Expansion of T cells induced by rDEV-dH5/H7 and vDEV. Four-week-old ducks were inoculated i.m. with two doses of 10 5 TCID 50 of rDEV-dH5/H7 or vDEV at a 3-week interval. The percentages of ( A ) CD3 + T cells, ( B ) CD3 + CD8 + T cells, and ( C ) CD3 + CD4 + T cells among the PBMCs were analyzed by flow cytometry. Data are presented as means and plotted with connecting lines. Each data point represents one sample value. The red triangles represent the time points of inoculation.

Article Snippet: The antibody cocktail included an FITC-conjugated anti-CD3 mAb (CD3-12; Abcam, ab11089) together with a PE-conjugated anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or a PE-conjugated anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479), which were labeled with a PE/R-phycoerythrin Conjugation Kit (Abcam, ab102918), respectively.

Techniques: Flow Cytometry

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Specific CD8 + T-cell responses induced by rDEV-dH5/H7 and vDEV. (A) Timeline of inoculation and sample testing. Arrows indicate inoculation, and red circles indicate bleeds, tests, and post-vaccination time points. ( B ) Frequency of specific CD3 + CD8 + T cells expressing IFN-γ + induced by DEV virion. ( C–F ) Frequency of HA-specific CD3 + CD8 + T cells expressing IFN-γ + . PBMCs were stimulated with single inactivated virus antigens: GZ/S4184 (H5N6), LN/SD007 (H5N1), GX/SD098 (H7N9), or a mixture of antigens of these three influenza virus antigens; CD3 + CD8 + IFN-γ + T cells were then quantified. ( G and H ) Uncorrelated control. CD3 + CD8 + IFN-γ + T cells were quantified after PBMCs were stimulated with WSN (H1N1) or La Sota (NDV). Data are presented as means in histograms. Each data point represents one sample value. Statistical significance: * P < 0.05, ** P <0.01, and *** P < 0.001. P values were determined using a two-tailed unpaired Student’s t test. ( I ) Representative gating strategy for HA-specific CD3 + CD8 + T cells expressing IFN-γ + detected at 31 days post-prime vaccination, following stimulation with the influenza virus antigens mixture. RPMI 1640 and PMA plus ionomycin served as negative and positive controls, respectively.

Article Snippet: The antibody cocktail included an FITC-conjugated anti-CD3 mAb (CD3-12; Abcam, ab11089) together with a PE-conjugated anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or a PE-conjugated anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479), which were labeled with a PE/R-phycoerythrin Conjugation Kit (Abcam, ab102918), respectively.

Techniques: Expressing, Virus, Control, Two Tailed Test

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Expansion of T cells induced by rDEV-dH5/H7 and vDEV. Four-week-old ducks were inoculated i.m. with two doses of 10 5 TCID 50 of rDEV-dH5/H7 or vDEV at a 3-week interval. The percentages of ( A ) CD3 + T cells, ( B ) CD3 + CD8 + T cells, and ( C ) CD3 + CD4 + T cells among the PBMCs were analyzed by flow cytometry. Data are presented as means and plotted with connecting lines. Each data point represents one sample value. The red triangles represent the time points of inoculation.

Article Snippet: The PBMCs were plated in 1.5 mL centrifuge tubes (10 6 cells/tube) and fixed with 100 μL of 0.3% paraformaldehyde for 20 min at room temperature and subsequently stained for 30 min at room temperature with either mouse anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or mouse anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479) diluted in PBS containing 0.05% Tween-20.

Techniques: Flow Cytometry

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Specific CD8 + T-cell responses induced by rDEV-dH5/H7 and vDEV. (A) Timeline of inoculation and sample testing. Arrows indicate inoculation, and red circles indicate bleeds, tests, and post-vaccination time points. ( B ) Frequency of specific CD3 + CD8 + T cells expressing IFN-γ + induced by DEV virion. ( C–F ) Frequency of HA-specific CD3 + CD8 + T cells expressing IFN-γ + . PBMCs were stimulated with single inactivated virus antigens: GZ/S4184 (H5N6), LN/SD007 (H5N1), GX/SD098 (H7N9), or a mixture of antigens of these three influenza virus antigens; CD3 + CD8 + IFN-γ + T cells were then quantified. ( G and H ) Uncorrelated control. CD3 + CD8 + IFN-γ + T cells were quantified after PBMCs were stimulated with WSN (H1N1) or La Sota (NDV). Data are presented as means in histograms. Each data point represents one sample value. Statistical significance: * P < 0.05, ** P <0.01, and *** P < 0.001. P values were determined using a two-tailed unpaired Student’s t test. ( I ) Representative gating strategy for HA-specific CD3 + CD8 + T cells expressing IFN-γ + detected at 31 days post-prime vaccination, following stimulation with the influenza virus antigens mixture. RPMI 1640 and PMA plus ionomycin served as negative and positive controls, respectively.

Article Snippet: The PBMCs were plated in 1.5 mL centrifuge tubes (10 6 cells/tube) and fixed with 100 μL of 0.3% paraformaldehyde for 20 min at room temperature and subsequently stained for 30 min at room temperature with either mouse anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or mouse anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479) diluted in PBS containing 0.05% Tween-20.

Techniques: Expressing, Virus, Control, Two Tailed Test

Journal: NPJ Parkinson's Disease

Article Title: Delphinidin modulates neuroinflammation and behavioral deficits in a Parkinson’s disease mouse model

doi: 10.1038/s41531-025-01244-0

Figure Lengend Snippet: a Quantification of CD4 + cells in the SN. Statistical analysis by one-way ANOVA followed by Tukey’s multiple comparisons test: F(3, 12) = 2.156, P = 0.1464, n-numbers: EV veh = 3, EV del = 4, hαSYN veh = 5, hαSYN del = 4 b Quantification of CD8 + cells in the substantia nigra (SN). Statistical analysis by one-way ANOVA followed by Tukey’s multiple comparisons test: F(3, 12) = 5.524, P = 0.0129, n-numbers: EV veh = 3, EV del = 4, hαSYN veh = 5, hαSYN del = 4 c Quantification of CD4 + cells in the striatum. Statistical analysis by one-way ANOVA followed by Tukey’s multiple comparisons test: F(3, 11) = 1.215, P = 0.3501, n-numbers: EV veh = 3, EV del = 4, hαSYN veh = 4, hαSYN del = 4 d Quantification of CD8 + cells in the striatum. Statistical analysis by one-way ANOVA followed by Tukey’s multiple comparisons test: F(3, 11) = 0.2133, P = 0.8851, n-numbers: EV veh = 3, EV del = 4, hαSYN veh = 4, hαSYN del = 4 e Representative IF images of CD8 + CD122 + cells in the SN. Scale bar: 50 µm f Quantification of CD8 + CD122 + cells in the SN – as indicated by TH + cells. Statistical analysis by one-tailed multiple t-tests, only showing significant results, P = 0.0425 (hαSYN veh vs. hαSYN del ), P = 0.0270 (EV veh vs. hαSYN del ), n-numbers: EV veh = 6, EV del = 6, hαSYN veh = 6, hαSYN del = 6; *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001. Data are shown as min-to-max and mean (+).

Article Snippet: Subsequently, sections were incubated overnight with rat anti-mouse CD4 (1:1000, Bio-Rad, cat. #MCA1767; RRID:AB_322769), rat anti-mouse CD8 (1:500, Bio-Rad, cat. #MCA609G; RRID:AB_321407), rat anti-mouse CD11b (1:100, Bio-Rad, cat. #MCA711; RRID:AB_321292), or rabbit anti-TH antibodies.

Techniques: One-tailed Test