Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: A Confirmation of CD8 + depletion by flow cytometry. Gating strategy: Supplementary Fig. . B – K MC38 tumor-bearing mice received 2 Gy 90 Y-, 177 Lu-, or 225 Ac-NM600 + ICI (days −3/0/3) + αCD8 (anti-CD8 on days −5/0/5), RPT + ICI, RPT alone, or αCD8 alone. Effects of CD8 + depletion on tumor growth ( B – H ) and overall survival ( I – K ) are shown. L Flow cytometry treatment scheme. MC38 tumor-bearing mice were randomized to receive either 2 Gy 90 Y-, 177 Lu-, or 225 Ac-NM600 + ICI (days −3/0/3), ICI alone, or untreated control (No Tx). M , N Tumors were harvested on days −3, 4, 8, or 11 and dissociated. The tumor immune infiltrate was analyzed by flow cytometry. Gating strategy: Supplementary Fig. . O Co-culture experiment scheme. Created https://BioRender.com/22ha1p4 . P , Q TNF and IFNγ expression were measured by intracellular flow cytometry staining and analysis of peripheral CD8 + effector memory cells (CD8 + CD44 + CD62L − ) harvested from naïve C57BL/6 mice or treated MC38 tumor-bearing mice following 24 h of co-culture with MC38 or no tumor cells. Gating strategy: Supplementary Fig. . A – H ; M , N ; P , Q Results of one experiment. I – K Results of two independent experiments. A N = 15: depleted; n = 7: IgG2b control. B – H N = 7: 90 Y-, 177 Lu-, 225 Ac-NM600 ± ICI; n = 5: 90 Y-, 177 Lu-, 225 Ac-NM600 + ICI + αCD8, αCD8. I – K N = 17: 90 Y-, 177 Lu-, 225 Ac-NM600 ± ICI; n = 10: αCD8; n = 5: 90 Y-, 177 Lu-, 225 Ac-NM600 + ICI + αCD8. M , N N = 5/treatment group and timepoint except n = 4: 90 Y-NM600 + ICI Day 4; 90 Y-, 177 Lu-, 225 Ac-NM600 + ICI Day 11. P , Q N = 5: naïve, 2 Gy 90 Y-NM600 + ICI; n = 6: ICI, 2 Gy 177 Lu-NM600 + ICI; n = 7: 2 Gy 225 Ac-NM600 + ICI. Unpaired two-tailed t-test was used to compare CD8 + frequency between groups. One-way ANOVA with Tukey’s HSD post hoc test was used to compare flow cytometry results between treatment groups for each marker and timepoint and TNF or IFNγ expression between treatment groups within each co-culture condition. Linear mixed models were used to compare tumor volumes over time between various treatment groups. Statistical testing of the pairwise contrasts was adjusted for multiple comparisons using Tukey’s method. Log-rank test was used to compare survival. Error bars are SEM. O Created in BioRender. Jin, W. (2026) https://BioRender.com/22ha1p4 .

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Flow Cytometry, Control, Co-Culture Assay, Expressing, Staining, Two Tailed Test, Marker

Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: MC38 tumor-bearing mice received IgG2b control on days −5/0/5 (IgG2b), or 0, 0.2 Gy (0.4625 kBq), 2 Gy (4.625 kBq), 8 Gy (18.5 kBq) 225 Ac-NM600 on day 1 ± ICI on days −3/0/3. A Body weights. B Complete blood counts on day 23. WBC white blood cells, LYM lymphocytes, PLT platelets, HGB hemoglobin, RBC red blood cells. C TNF and IFNγ on peripheral CD8 + effector memory cells (CD8 + CD44 + CD62L − ) from naïve or MC38 tumor-bearing mice receiving the indicated treatment and following 24 h of co-culture with MC38 or no tumor cells (from Fig. ). D – L MC38 tumor-bearing mice received 8 Gy 225 Ac-NM600 + ICI, 8 Gy 225 Ac-NM600, ICI, or were untreated (No Tx). D Experimental scheme. Created with BioRender.com. Tumor growth ( E ) and overall survival ( F ) are shown. G Complete response (CR) rates ( H ) and percent of CR mice and naïve controls rejecting rechallenge with MC38 cells are shown. I – K Flow cytometry of blood from CR (MC38, Ac + ICI) or naïve mice (MC38, naïve). L Flow cytometry of splenocytes from CR 225 Ac-NM600 + ICI mice that did not (Ac + ICI tumor) or did reject rechallenge (Ac + ICI no tumor), CR ICI mice (ICI), naïve C57BL/6 mice following IV injection of MC38 cells (MC38 IV), and completely naïve C57BL/6 mice (Naïve). Gating strategies: Supplementary Figs. – . A , B ; C ; E – L Results of one experiment. A N = 7: all groups. B N = 5: naïve, 2 Gy 90 Y-NM600 + ICI; n = 6: ICI, 2 Gy 177 Lu-NM600 + ICI; n = 7: 0.2, 2, 8 Gy 225 Ac-NM600 + ICI. C N = 5: naïve; n = 6: ICI; n = 7: 0.2, 2, 8 Gy 225 Ac-NM600 + ICI. E – G N = 5/treatment group. H N = 5: 225 Ac-NM600 + ICI, naïve; n = 3: ICI. I – K N = 4/treatment group. L N = 4: MC38 IV, Naïve; n = 3: ICI; n = 2: Ac + ICI tumor, Ac + ICI no tumor. One-way ANOVA with Tukey’s HSD post hoc test was used to compare TNF or IFNγ expression, blood counts, CR rates, rechallenge rejection rates, and spleen flow cytometry. Unpaired two-tailed t-test was used to compare blood flow cytometry. Tumor volumes were compared using linear mixed models adjusted for multiple comparisons using Tukey’s method. Log-rank test was used to compare survival. Error bars are SEM. D Created in BioRender. Berg, T. (2026) https://BioRender.com/llbyoho, .

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Control, Co-Culture Assay, Flow Cytometry, IV Injection, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: Paired TCR sequencing analysis performed on samples from B78 tumor-bearing mice used for single cell RNA sequencing. A – H Data presented are from samples from experiment depicted in Fig. . A UMAP plots of T cells stratified by sample. B TCR clonotype expansion ranges. C UMAP plots of T cells stratified by T cell subtype. D Clonal expansion by treatment group and T cell subtype. E D50 score by treatment group and T cell subtype. F Relative abundance of clonal indices for each treatment group and T cell subtype. G UpSet plot showing shared clonotypes between T cell subtypes for the 225 Ac-NM600 + ICI sample. Red box demarcates CD8 + effector and memory subsets; red arrow demarcates # of shared clonotypes between CD8 + effector and memory subsets (=11). H UpSet plots showing shared clonotypes between T cell subtypes for the 90 Y-, 177 Lu-, EBRT + ICI, and ICI alone samples. For each plot, red arrow demarcates # of shared clonotypes between CD8+ effector and memory subsets (=7 for each sample). A – H Results of one experiment. 3 mice/treatment group pooled into one sample/treatment group for TCR sequencing analysis. Numbers of cells analyzed provided in Supplementary Table . Number of cells analyzed: 90 Y-NM600 + ICI, n = 408; 177 Lu-NM600 + ICI, n = 993; 225 Ac-NM600 + ICI, n = 854; EBRT + ICI, n = 285; ICI, n = 977.

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Sequencing, Single Cell, RNA Sequencing

Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: A – D Data presented are from samples from experiment depicted in Fig. . A Pathway enrichment results for CD8 + effector T cells. The bluer the pathway, the lower the adjusted P, and thus the more significantly the pathway is altered. Differential gene expression heat map for interferon γ ( B ) and interferon α ( C ) response genes for CD8 + effector T cells. The color of each box represents the log 2 (fold change) of gene expression of 225 Ac-, 177 Lu-, 90 Y-NM600, or EBRT + ICI compared to ICI alone. D Incidence matrix of differential gene expression between treatment groups of the following: comparing cells where the shared clonotypes between effector and memory CD8 + T cells are between 7 and 11 (as shown in Fig. ), and they are amplified. Red arrow marks interferon-inducible gene, Ifi27l2a , which is upregulated in these shared clonotype CD8 + cell populations treated with 225 Ac-NM600 + ICI over each treatment group. A – D Results of one experiment. 3 mice/treatment group pooled into one sample/treatment group for single cell RNA sequencing TCR sequencing analysis. Benjamini–Hochberg method was used to correct for multiple comparison. Numbers of cells analyzed provided in Supplementary Table A – C 90 Y-NM600 + ICI, n = 6067; 177 Lu-NM600 + ICI, n = 9808; 225 Ac-NM600 + ICI, n = 6503; EBRT + ICI, n = 7045; ICI, n = 11,249 and Supplementary Table D 90 Y-NM600 + ICI, n = 123; 177 Lu-NM600 + ICI, n = 151; 225 Ac-NM600 + ICI, n = 45; EBRT + ICI, n = 36; ICI, n = 100.

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Gene Expression, Amplification, Single Cell, RNA Sequencing, Sequencing, Comparison