anti cd8α  (Bio X Cell)

Journal: bioRxiv

Article Title: Addition of chemotherapy to radiotherapy promotes progenitor-exhausted CD8⁺ T-cell clonal dominance in head and neck cancer

doi: 10.64898/2026.03.10.710795

Figure Lengend Snippet: Experiments performed in the mEER model. A. Tumour growth curves across indicated treatment conditions. B. Survival curves across indicated treatment conditions (A-B: Vehicle n=11, Cisplatin n=11, RT + vehicle n=11, CRT n=12; 2 independent experiments). C. Absolute change of CD8 + T-cell counts/mg tumour vs average of timematched controls across indicated timepoints post-RT as measured by flow cytometry (RT d3 n = 9, RT d7 n = 15, RT d10 n = 17, RT d14 n = 6; 6 independent experiments of 2 consecutive timepoints). D. CD8 + T-cells/ mm 2 tumour at 14 days post-RT by immunohistochemistry, necrotic regions excluded, quantified by QuPath v. 0.3.0. (Vehicle n=13, RT, n=16, CRT n=11; 3 independent experiments). E. CD8 + T-cell count/ mg of tumour by flow cytometry (Vehicle n=10, Cisplatin n=11, RT n=10, CRT n=11; 2 independent experiments). F. Percentage of PD-1 hi CD8 + T-cells by flow cytometry (Vehicle n=11, Cisplatin n=12, RT n=11, CRT n=12; 2 independent experiments). G-H: Tumour growth curves of ⍺CD8 depletion experiments, with mice receiving either RT (G) or CRT (H) (Vehicle + isotype n=6, RT + isotype n=6, RT + ⍺CD8 n = 5, CRT + isotype n = 6, CRT + ⍺CD8 n = 6; data from 1 experiment). Results are means ± SEM and n denotes mice per group. Parametric statistics were only applied to normally distributed data (Shapiro-Wilk test). P-values shown were determined by the log-rank (Mantel–Cox) (B), Kruskal Wallis test with Dunn’s multiple comparisons test (C-E), Ordinary one-way ANOVA with Tukey’s multiple comparisons test (F) and Two-tailed Unpaired t test (H).

Article Snippet: CD8 + T-cell depletion was performed using an anti-CD8α antibody (loading dose 400 μg, maintenance 200 μg twice weekly; clone: 2.43, BioXCell) or rat IgG2b isotype control (clone: 1-2, BioXCell) for up to 10 doses.

Techniques: Flow Cytometry, Immunohistochemistry, Cell Characterization, Two Tailed Test

Journal: bioRxiv

Article Title: Addition of chemotherapy to radiotherapy promotes progenitor-exhausted CD8⁺ T-cell clonal dominance in head and neck cancer

doi: 10.64898/2026.03.10.710795

Figure Lengend Snippet: A. Schematic of the treatment timeline in Nr4a3-Tocky mEER tumour-bearing mice. Created with Biorender.com. B-F: UMAP analysis of CD8 + tumour-infiltrating lymphocytes (TILs), results concatenated from 30 mice (Vehicle n=7, Cisplatin n=8, RT + vehicle n=7, CRT n=8; 1 experiment). (B) UMAP plot showing distribution of clusters 1–8 across treatment conditions. (C) Selected clusters 2, 3, 4 and 5 showing significant differences between treatment groups (Vehicle n=7, Cisplatin n=8, RT + vehicle n=7, CRT n=8). (D) Timer fluorescence patterns within clusters 2, 3, 4 and 5. Cluster 3 is defined by a persistent-arrested (PAt) Tocky profile with high activation and immune checkpoint expression. (E) Heatmap overlays of Tocky “angle” (antigen engagement), “intensity” (TCR signalling strength). (F) Histograms illustrating marker expression by cluster. G. Percentage change in PAt CD8 + T-cells (% ΔPAt) in RT + vehicle and CRT groups versus the average of vehicle group (RT + vehicle n=11, CRT n=12; 2 independent experiments). Results are means ± SEM and n denotes mice per group. Parametric statistics were only applied to normally distributed data (Shapiro-Wilk test). P-values shown were determined by Ordinary one-way ANOVA with Tukey’s multiple comparisons test (E: cluster 2-3), Kruskal Wallis test with Dunn’s multiple comparisons test (E: cluster 4-5) and Two-tailed Unpaired t test (G). Significant outliers were removed via ROUT test (Q = 1%). All data are representative of two independent experiments.

Article Snippet: CD8 + T-cell depletion was performed using an anti-CD8α antibody (loading dose 400 μg, maintenance 200 μg twice weekly; clone: 2.43, BioXCell) or rat IgG2b isotype control (clone: 1-2, BioXCell) for up to 10 doses.

Techniques: Fluorescence, Activation Assay, Expressing, Marker, Two Tailed Test

Journal: bioRxiv

Article Title: Addition of chemotherapy to radiotherapy promotes progenitor-exhausted CD8⁺ T-cell clonal dominance in head and neck cancer

doi: 10.64898/2026.03.10.710795

Figure Lengend Snippet: A. Experimental schema: Single-cell analysis was performed on tumour-infiltrating lymphocytes (TILs) from tumours (Tum) or TDLN harvested from mEER-bearing Nr4a3-Tocky mice treated with vehicle (Veh), RT or CRT. Cells were sorted according to Tocky antigen-engagement profile prior to integrated single-cell analyses (Vehicle n=12, RT n=12, CRT n=12; 1 experiment). B-H: Cells were subject to UMAP clustering based on Abseq expression profile. (B) UMAP by sample tag. (C) Cell annotation of T-cell subtype: CD4, CD8, DN (double negative) and DP (double positive). (D) Functional T-cell subtype annotation based on expression markers: T N (naïve, CD62L + ), T CM (central memory, CD62L + , CD44 + ), T EM (effector memory, CD62L - CD44 + ). (E) Heatmap of selected Abseq marker expression overlaid on UMAP. (F) Dot plot of Abseq marker expression in TILs by treatment. (G) Distribution of Tocky populations: “persistent” (pers), “arrested” (arr) and “timer negative” (neg) within the UMAP space. (H) Dot plots of Abseq marker expression across different Tocky populations in TILs.

Article Snippet: CD8 + T-cell depletion was performed using an anti-CD8α antibody (loading dose 400 μg, maintenance 200 μg twice weekly; clone: 2.43, BioXCell) or rat IgG2b isotype control (clone: 1-2, BioXCell) for up to 10 doses.

Techniques: Single-cell Analysis, Single Cell, Expressing, Functional Assay, Marker

Journal: bioRxiv

Article Title: Addition of chemotherapy to radiotherapy promotes progenitor-exhausted CD8⁺ T-cell clonal dominance in head and neck cancer

doi: 10.64898/2026.03.10.710795

Figure Lengend Snippet: Single-cell TCR sequencing was performed on sorted tumour-infiltrating lymphocytes (TILs) from tumours (Tum) or TDLN harvested from mEER-bearing mice treated with vehicle (Veh), RT or CRT (Vehicle n=12, RT n=12, CRT n=12, 1 experiment). A. Distribution of T-cell clonal expansion within UMAP space, categorised by clone frequency within the total TCR repertoire: hyperexpanded (>20 and ≤100), large (>5 and ≤20), medium (>2 and ≤5), small (>1 and ≤2) and single (>0 and ≤1). Cells without information on paired TCR α (alpha) and β (beta) chain CDR3 (Complementarity-Determining Region 3) sequence information are denoted by “NA”. B. Three CDR3 sequences which were hyperexpanded. C. Distribution of categories of clonal expansion within UMAP space by sample type and treatment group. D. Categories of clonal expansion by sample type and treatment group by proportion of cells E. Clonal diversity dotplots comparing various diversity indices between Tum_RT and Tum_CRT: Shannon; Inverse (Inv) Simpson; Chao1 (Chao) Abundance-based Coverage Estimator (ACE) and Inv Pielou. F. Proportions of T-cell type (CD4+, CD8+, double negative [DN], double positive [DP]) by categories of clonal expansion. G. Abseq expression marker profile across different clonal expansion categories.

Article Snippet: CD8 + T-cell depletion was performed using an anti-CD8α antibody (loading dose 400 μg, maintenance 200 μg twice weekly; clone: 2.43, BioXCell) or rat IgG2b isotype control (clone: 1-2, BioXCell) for up to 10 doses.

Techniques: Single Cell, Sequencing, Expressing, Marker

Journal: bioRxiv

Article Title: Addition of chemotherapy to radiotherapy promotes progenitor-exhausted CD8⁺ T-cell clonal dominance in head and neck cancer

doi: 10.64898/2026.03.10.710795

Figure Lengend Snippet: Single-cell analysis was performed on sorted tumour-infiltrating lymphocytes (TILs) from tumours (Tum) or TDLN harvested from mEER-bearing mice treated with vehicle (Veh), RT or CRT (Vehicle n=12, RT n=12, CRT n=12, 1 experiment). A. Distribution of Tocky populations within UMAP space according to categories of clonal expansion: “new”, “persistent” (pers), “arrested” (arr) and “timer negative” (neg). B. Proportions of cells undergoing clonal expansion by Tocky population. C-E: Abseq marker expression profile of various TILs subsets: (C) Clonal (ζ 5) and “persistent” CD8 + TILs by treatment type, (D) Clonal (ζ 5) CD8 + TILs from mice treated with CRT (Tum_CRT) by Tocky population and (E) Clonal (ζ 5) CD4 + TILs treated with CRT (Tum_CRT) by Tocky population.

Article Snippet: CD8 + T-cell depletion was performed using an anti-CD8α antibody (loading dose 400 μg, maintenance 200 μg twice weekly; clone: 2.43, BioXCell) or rat IgG2b isotype control (clone: 1-2, BioXCell) for up to 10 doses.

Techniques: Single-cell Analysis, Marker, Expressing