Journal: Immunity

Article Title: Central memory CD8 + T cells derive from stem-like Tcf7 hi effector cells in the absence of cytotoxic differentiation.

doi: 10.1016/j.immuni.2020.09.005

Figure Lengend Snippet: Figure 1. Effector-Stage Tcf7hi CD8+ T Cells Resemble Central Memory Cells B6 (CD45.1/2) mice were adoptively transferred with Tcf7GFP P14 cells (CD45.2) and infected with LCMV WE. (A) Tcf7GFP expression by splenic P14 cells at the indicated time points post-infection (p.i.).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Peptide: Ovalbumin amino acids 257-264 (OVA) (SIINFEKL) P. Romero, UNIL N/A Peptide: KL-SLP (KKKKKLEQLEAAYSIINFEKL) GenScript, NJ N/A Percoll GE Heathcare Cat# 17-0891-01 Polybrene Sigma-Aldrich Cat# TR-1003-G Recombinant human IL-2 Glaxo IMB, Genève, Switzerland gift from N. Rufer Sunflower seed oil Sigma-Aldrich Cat# S5007 Tamoxifen Sigma-Aldrich Cat# T5648 TDE1, Tagment DNA Enzyme Illumina Cat# 15027865 Trizol Life Technlogies Cat# 15596026 2xTD buffer Illumina Cat# 15027866 7-AAD (Viability dye) Biolegend Cat# 420404 Critical Commercial Assays Tumor Dissociation Kit Miltenyi Biotec Cat# 130-096-730 Mouse CD8+ T cell enrichment kit StemCell Technologies Cat# 19853 Direct–zol RNA Mini Prep Zymo Research Cat# R2050 Intracellular Fix & Perm Buffer set eBiosciences Cat# 88-8824 FoxP3/Transcription factor staining buffer set eBiosciences Cat# 00-5523 SMART-Seq v4 Ultra Low Input RNA reagents Clontech Cat# 634888 Illumina Nextera XT DNA Library reagents Illumina Cat# 15032354 MACS CD8+ positive selection Miltenyi Biotec Cat# 130-116-478 Annexin V - APC Apoptosis Detection Kit eBioscience RRID:AB_2575165 SuperScript III First-Strand Synthesis System ThermoFisher Scientific Cat# 18080051 KAPA SYBR FAST qPCR Kit Master Mix Kapabiosystems Cat# KR0389 Dynabeads Mouse T-Activator CD3/CD28 ThermoFisher Scientific Cat# 11452D CellTrace Violet Cell Proliferation Kit ThermoFisher Scientific Cat# C34557 MinElute PCR Purification kit QIAGEN Cat# 28004 Illumina’s Unique Dual (UD) Indexes Illumina Cat# 20027213 NEBNext High-Fidelity 2X PCR Master Mix New England Biolabs Cat# M0541 AMPure XP magnetic beads Beckman Coulter Cat# A63880 Zombie Aqua Fixable Viability kit Biolegend Cat# 423101 Deposited Data RNA-seq data This study GEO: GSE144383 ATAC-seq data This study GEO GSE144383 Experimental Models: Cell Lines Mouse: RMA RRID:CVCL_J385 Experimental Models: Organisms/Strains Mouse: C57BL/6 (B6) (CD45.2) Charles River Laboratoies Strain 027 Mouse: B6.SJL-Ptprc < a > (B6 CD45.1) Jackson Lab Strain 002014; RRID:MGI:6200621 Mouse: B6; D2-Tg(TcrLCMV)327Sdz P14 T cell receptor (TCR) transgenic (CD45.2) (Pircher et al., 1989) RRID:MGI:3810256 Mouse: B6.129-Tm(Tcf7)Cle (Tcf7 / ) (CD45.2) Verbeek et al., 1995 RRID:MGI:4360712 (Continued on next page) Immunity 53, 1–16.e1–e11, November 17, 2020 e3

Techniques: Infection, Expressing

Journal: Immunity

Article Title: Central memory CD8 + T cells derive from stem-like Tcf7 hi effector cells in the absence of cytotoxic differentiation.

doi: 10.1016/j.immuni.2020.09.005

Figure Lengend Snippet: Figure 5. Vaccination of Mice and Humans Generates Effector-Phase Tcf1hi CD8+ T Cells (A–H) B6 Tcf7GFP mice were vaccinated as indicated with a modified Ovalbumin peptide and Pam3CSK4 in Montanide (Ova) or with Pam3CSK4 in Montanide (Ø). (B, C, and E) Abundance and (D and F) Tcf7GFP and CD62L expression by KbOva+ CD8+ T cells in the blood on d21 and d35 (B–D) and in the spleen on d35 post- vaccination (E–G). (H) IL-2 and TNF-a production by d35 splenic IFN-g+ cells. (I–K) Healthy volunteers received a yellow fever vaccine, and peripheral blood CD8+ T cells were analyzed 14 days later. (I) TCF1 expression by A2/LLW tetramer+

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Peptide: Ovalbumin amino acids 257-264 (OVA) (SIINFEKL) P. Romero, UNIL N/A Peptide: KL-SLP (KKKKKLEQLEAAYSIINFEKL) GenScript, NJ N/A Percoll GE Heathcare Cat# 17-0891-01 Polybrene Sigma-Aldrich Cat# TR-1003-G Recombinant human IL-2 Glaxo IMB, Genève, Switzerland gift from N. Rufer Sunflower seed oil Sigma-Aldrich Cat# S5007 Tamoxifen Sigma-Aldrich Cat# T5648 TDE1, Tagment DNA Enzyme Illumina Cat# 15027865 Trizol Life Technlogies Cat# 15596026 2xTD buffer Illumina Cat# 15027866 7-AAD (Viability dye) Biolegend Cat# 420404 Critical Commercial Assays Tumor Dissociation Kit Miltenyi Biotec Cat# 130-096-730 Mouse CD8+ T cell enrichment kit StemCell Technologies Cat# 19853 Direct–zol RNA Mini Prep Zymo Research Cat# R2050 Intracellular Fix & Perm Buffer set eBiosciences Cat# 88-8824 FoxP3/Transcription factor staining buffer set eBiosciences Cat# 00-5523 SMART-Seq v4 Ultra Low Input RNA reagents Clontech Cat# 634888 Illumina Nextera XT DNA Library reagents Illumina Cat# 15032354 MACS CD8+ positive selection Miltenyi Biotec Cat# 130-116-478 Annexin V - APC Apoptosis Detection Kit eBioscience RRID:AB_2575165 SuperScript III First-Strand Synthesis System ThermoFisher Scientific Cat# 18080051 KAPA SYBR FAST qPCR Kit Master Mix Kapabiosystems Cat# KR0389 Dynabeads Mouse T-Activator CD3/CD28 ThermoFisher Scientific Cat# 11452D CellTrace Violet Cell Proliferation Kit ThermoFisher Scientific Cat# C34557 MinElute PCR Purification kit QIAGEN Cat# 28004 Illumina’s Unique Dual (UD) Indexes Illumina Cat# 20027213 NEBNext High-Fidelity 2X PCR Master Mix New England Biolabs Cat# M0541 AMPure XP magnetic beads Beckman Coulter Cat# A63880 Zombie Aqua Fixable Viability kit Biolegend Cat# 423101 Deposited Data RNA-seq data This study GEO: GSE144383 ATAC-seq data This study GEO GSE144383 Experimental Models: Cell Lines Mouse: RMA RRID:CVCL_J385 Experimental Models: Organisms/Strains Mouse: C57BL/6 (B6) (CD45.2) Charles River Laboratoies Strain 027 Mouse: B6.SJL-Ptprc < a > (B6 CD45.1) Jackson Lab Strain 002014; RRID:MGI:6200621 Mouse: B6; D2-Tg(TcrLCMV)327Sdz P14 T cell receptor (TCR) transgenic (CD45.2) (Pircher et al., 1989) RRID:MGI:3810256 Mouse: B6.129-Tm(Tcf7)Cle (Tcf7 / ) (CD45.2) Verbeek et al., 1995 RRID:MGI:4360712 (Continued on next page) Immunity 53, 1–16.e1–e11, November 17, 2020 e3

Techniques: Expressing

Journal: Immunity

Article Title: Central memory CD8 + T cells derive from stem-like Tcf7 hi effector cells in the absence of cytotoxic differentiation.

doi: 10.1016/j.immuni.2020.09.005

Figure Lengend Snippet: Figure 6. Tcf1 Is Essential for the Stemness of d8 Tcf7GFPhi CD8+ T Cells (A–D) B6 mice (CD45.1/2) were transplanted with WT or Tcf7/ (KO) Tcf7GFP P14 cells (CD45.2) and infected with LCMV WE. (A) Abundance of P14 cells in the spleen at d8 p.i. and (B) expression of Tcf7GFP. Phenotype (C) and cytokine production (D) by WT and KO Tcf7GFPhi P14 cells. (E–G) Recall response of sorted d8 WT and KO Tcf7GFPhi P14 cells. (E) Abundance of P14 cells in the spleen and (F) Tcf7GFP expression 8 days later (d8+8). (G) Abundance of secondary Tcf7GFPhi cells compared to input. Data are representative of 3 experiments with 4 mice per group (A–C), representative of 2 experiments with 5–6 mice per group (D), or compiled from 2 ex- periments with 5–7 mice per group (E–G). Mean ± SD are shown. Statistics: non-paired t test (A–D and F) or one-way ANOVA with Tukey’s test (E and G) with *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; and (ns) p > 0.05. See also Figure S7.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Peptide: Ovalbumin amino acids 257-264 (OVA) (SIINFEKL) P. Romero, UNIL N/A Peptide: KL-SLP (KKKKKLEQLEAAYSIINFEKL) GenScript, NJ N/A Percoll GE Heathcare Cat# 17-0891-01 Polybrene Sigma-Aldrich Cat# TR-1003-G Recombinant human IL-2 Glaxo IMB, Genève, Switzerland gift from N. Rufer Sunflower seed oil Sigma-Aldrich Cat# S5007 Tamoxifen Sigma-Aldrich Cat# T5648 TDE1, Tagment DNA Enzyme Illumina Cat# 15027865 Trizol Life Technlogies Cat# 15596026 2xTD buffer Illumina Cat# 15027866 7-AAD (Viability dye) Biolegend Cat# 420404 Critical Commercial Assays Tumor Dissociation Kit Miltenyi Biotec Cat# 130-096-730 Mouse CD8+ T cell enrichment kit StemCell Technologies Cat# 19853 Direct–zol RNA Mini Prep Zymo Research Cat# R2050 Intracellular Fix & Perm Buffer set eBiosciences Cat# 88-8824 FoxP3/Transcription factor staining buffer set eBiosciences Cat# 00-5523 SMART-Seq v4 Ultra Low Input RNA reagents Clontech Cat# 634888 Illumina Nextera XT DNA Library reagents Illumina Cat# 15032354 MACS CD8+ positive selection Miltenyi Biotec Cat# 130-116-478 Annexin V - APC Apoptosis Detection Kit eBioscience RRID:AB_2575165 SuperScript III First-Strand Synthesis System ThermoFisher Scientific Cat# 18080051 KAPA SYBR FAST qPCR Kit Master Mix Kapabiosystems Cat# KR0389 Dynabeads Mouse T-Activator CD3/CD28 ThermoFisher Scientific Cat# 11452D CellTrace Violet Cell Proliferation Kit ThermoFisher Scientific Cat# C34557 MinElute PCR Purification kit QIAGEN Cat# 28004 Illumina’s Unique Dual (UD) Indexes Illumina Cat# 20027213 NEBNext High-Fidelity 2X PCR Master Mix New England Biolabs Cat# M0541 AMPure XP magnetic beads Beckman Coulter Cat# A63880 Zombie Aqua Fixable Viability kit Biolegend Cat# 423101 Deposited Data RNA-seq data This study GEO: GSE144383 ATAC-seq data This study GEO GSE144383 Experimental Models: Cell Lines Mouse: RMA RRID:CVCL_J385 Experimental Models: Organisms/Strains Mouse: C57BL/6 (B6) (CD45.2) Charles River Laboratoies Strain 027 Mouse: B6.SJL-Ptprc < a > (B6 CD45.1) Jackson Lab Strain 002014; RRID:MGI:6200621 Mouse: B6; D2-Tg(TcrLCMV)327Sdz P14 T cell receptor (TCR) transgenic (CD45.2) (Pircher et al., 1989) RRID:MGI:3810256 Mouse: B6.129-Tm(Tcf7)Cle (Tcf7 / ) (CD45.2) Verbeek et al., 1995 RRID:MGI:4360712 (Continued on next page) Immunity 53, 1–16.e1–e11, November 17, 2020 e3

Techniques: Infection, Expressing

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) UMAP plot of 109,914 HNSCC TILs, including 106,667 CD8 T cells and 3,247 natural killer (NK) cells. (B) Heatmap displaying average Z-scored RNA expression per cluster. Notable genes, including ENTPD1 (CD39), CCR6 , and KLRC1 (NKG2A) are highlighted and boxed in red. (C) Representative plots of NKG2A and CCR6 expression on the total Tex population (CD39+ KLRG1-) for an HPV- and HPV+ HNSCC tumor. (D) Percentage of each Tex subset within the total Tex population (CD39+ KLRG1-) for Tex-Conv ( left , NKG2A- CCR6-), Tex-CCR6 ( middle , NKG2A- CCR6+), and Tex-KLR ( right , NKG2A+ CCR6-; n = 15 HPV-, 16 HPV+; Student’s t test). (E) Mean fluorescence intensity (MFI) of CD39 within each Tex subset and non-Tex cells (CD39-KLRG1-/+; n = 31; Tukey’s multiple comparisons test). (F) Ternary plot of the proportion of cells of each expanded Tex clonotype among the Tex clusters. Each dot represents an expanded Tex clonotype and is sized by the total cells in the clone. The left axis is the proportion of cells in the Tex-KLR cluster with 0 to 1 oriented from bottom to top. The right axis is the proportion of cells in the Tex-CCR6 cluster with 0 to 1 oriented from top to bottom. The bottom axis is the proportion of cells in the Tex-Conv cluster with 0 to 1 oriented from right to left. (G) Density plots of the individual cells fro each biased Tex clonotype category.

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: RNA Expression, Expressing, Fluorescence

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) UMAP plot demonstrating the transfer of cluster annotations from the reference CD8 T cell atlas ( left , Xue et al., 2024) to our HNSCC CD8+ TIL dataset ( right ). (B) Stacked bar plots showing the proportions of the transferred annotations within each of our annotated HNSCC CD8+ TIL clusters. (C) Dot plot of the top five marker genes per cluster (log 2 fold change > 1 and minimum percent of cells in which gene was detected > 50%). Genes of interest, including KLRC1 , CCR6 , and ENTPD1 , are highlighted in red.

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Marker

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Flow cytometry gating strategy for HNSCC patient tumor samples. (B-D) Percentage of (B) CD4 T cells of total T cells, (C) CD8 T cells among of T cells, and (D) exhausted CD8 T cells of CD8 T cells (n = 15 HPV-, 16 HPV+; Student’s t test; box plot whiskers show the full range).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Flow Cytometry

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Total number of CD8+ TILs analyzed per HNSCC patient sample. The fill of the bar indicates number of cells with a TCR captured by scTCR-seq. (B) Number of expanded Tex clonotypes biased towards each Tex cluster per patient. (C-D) Cellular composition of the top expanded clonotypes among the (C) Tex population or (D) the total CD8+ TIL population across clusters for all patients with > 600 cells with a TCR.

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques:

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Representative low-magnification images of pan-cytokeratin (PanCK) and TP63 ( top ) in addition to CD8 and CD3E ( bottom ). (B) High-magnification views of the boxed region in (A) for CD8 and NKG2A ( top left ), CD8 ( middle left ), NKG2A ( bottom left ), PanCK and TP63 ( top right ), TOX and PD-1 ( middle right ), and GZMB ( bottom right ). NKG2A+ Tex cells are indicated with a filled arrow, and NKG2A- Tex cells are indicated with an empty arrow. The circle indicates a region of interest. (C) Percentage of non-exhausted ( left ) and exhausted ( right ) cells of the total cells in each 10 µm bin spanning -250 µm to +250 µm relative to the tumor-stroma interface. The lines are the patient-specific locally estimated scatterplot smoothing fits, and shaded ribbons are the 95% confidence intervals for the fit of the line. (D) Percentage of NKG2A+ cells within each CD8 T cell subset (n = 11 samples; Wilcoxon rank-sum test). (E) Percentage of NKG2A+ Tex-KLR cells of total Tex cells in the intra-tumoral and peri-tumoral regions (n = 10 tumors with ≥ 100 cells per region; Wilcoxon signed-rank test).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques:

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Schematic of the avidity-modulated UM-SCC47 cell line models, including HLA-A2 OE with peptide pulsing (500 nM peptide) or natural E7 11-20 presentation ( left ) and the wildtype and mutant E7 11-20 SCT cell lines ( right ). (B) Representative histograms of CD3, CD137, and NKG2A expression on TCR-transduced CD8 T cells, normalized to the mode, following 48 h co-culture with each UM-SCC47 cell line. Data for donor LRS071724 is shown. (C) Representative flow cytometry plots of NKG2A and CD137 expression on TCR-transduced CD8 T cells following 48 h co-culture with each UM-SCC47 cell line. (D-F) Quantification of (D) CD3 mean fluorescence intensity (MFI) relative to T cells alone, (E) CD137 MFI relative to T cells alone, and (F) percentage of NKG2A+ cells among TCR-transduced CD8 T cells (n = 4 donors; repeated-measures Sidak’s multiple comparisons test; donors LRS050924, LRS071724, LRS081224, and LRS081524 were used).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Mutagenesis, Expressing, Co-Culture Assay, Flow Cytometry, Fluorescence

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Flow cytometry gating strategy for analysis of TCR-transduced CD8 T cells following in vitro co-culture. (B) HLA-A2 expression on the UM-SCC47 cell lines displayed as histograms ( left ) and quantified as mean fluorescence intensity (MFI; right ). (C) IL-12p70 concentration (pg mL -1 ) following 48 h of culture in the indicated condition (n = 4 donors; donors 85, 267, 481, and 948 were used).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Flow Cytometry, In Vitro, Co-Culture Assay, Expressing, Fluorescence, Concentration Assay

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Schematic of the avidity-modulated HEK293T cell line (naturally HLA-A2+) models, including HLA-A2 OE, HPV16 E7 11-20 peptide pulsing (500 nM peptide), and HPV16 E7 OE ( left ), and the wildtype and mutant E7 11-20 SCT cell lines ( right ). (B) Representative histograms of CD3, CD137, and NKG2A expression on TCR-transduced CD8 T cells, normalized to the mode, for each co-culture condition. Data for donor LRS071724 is shown. (C) Representative flow cytometry plots of NKG2A and CD137 expression on TCR-transduced CD8 T cells following 48 h co-culture with each HEK293T cell line. (D-F) Quantification of (D) CD3 mean fluorescence intensity (MFI) relative to T cells alone, (E) CD137 MFI relative to T cells alone, and (F) percentage of NKG2A+ cells among TCR-transduced CD8 T cells (n = 3 donors; repeated-measures Sidak’s multiple comparisons test; donors LRS072323, LRS050924, and LRS071724 were used).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Mutagenesis, Expressing, Co-Culture Assay, Flow Cytometry, Fluorescence

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Representative flow cytometry plots of NKG2A and CD137 expression on TCR-transduced CD8 T cells following 48 h culture in the indicated condition. FK506 was used at 5 nM, and IL-12p70 was used at 10 ng mL -1 . The vehicle control was 0.0005% ethanol. Data for donor LRS081324 is shown. (B-C) Percentage of CD137+ ( left ) and NKG2A+ ( right ) TCR-transduced CD8 T cells in the indicated treatments for (B) T cells alone and stimulation with anti-CD3/anti-CD28 and (C) co-culture with UM-SCC47 target cell lines (n = 3 donors, repeated-measures Tukey’s multiple comparisons test). (D) Percentage of UM-SCC47 target cells killed after 48 h co-culture. Values were mean-centered across experiments for visualization (n = 7 replicates from 4 unique donors, repeated-measures Tukey’s multiple comparisons test).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Flow Cytometry, Expressing, Control, Co-Culture Assay

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Principal component analysis (PCA) plot of the bulk RNA-seq data from isolated TCR-transduced CD8 T cells following 48 hours of co-culture with each UM-SCC47 cell line and treatment condition (n = 4 donors per condition). (B-C) Log 2 normalized counts for (B) TNFRSF9 and (C) KLRC1 . (D) Heatmap of activation-induced genes defined as genes upregulated in the wildtype E7 11-20 SCT or HLA-A2 OE + IL-12p70 condition relative to the empty vector control. Genes are sorted by the log 2 fold change of gene expression in the comparison of HLA-A2 OE + IL-12p70 over wildtype E7 11-20 SCT. (E) Log 2 fold change of the differentially expressed genes for HLA-A2 OE + IL-12p70 versus HLA-A2 OE ( y-axis ) and wildtype E7 11-20 SCT versus HLA-A2 OE ( x-axis ). Points are colored by their respective k-means cluster from the heatmap in (D) . The diagonal lines indicate the 0.75 delta log 2 fold change threshold between the two comparisons. Colored overlays and labels denote the genes selected for the IL-12-specific, shared, and avidity-specific gene sets.

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: RNA Sequencing, Isolation, Co-Culture Assay, Activation Assay, Plasmid Preparation, Control, Gene Expression, Comparison

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Module scores of the bulk RNA-seq gene sets. Datapoints are the median module score per patient among the cells in the cluster (n = 30-32 patients per cluster; Dunnett’s multiple comparisons test). (B) Mean Z-scored RNA expression per cluster of the genes in the bulk RNA-seq gene sets. Genes are sorted by the delta Z-score of the Tex-KLR cluster minus the mean of the Tex-Conv and Tex-CCR6 clusters. Only genes detected in > 5% of cells in at least one cluster are shown. (C) Avidity-specific gene set module score of the cells within representative expanded Tex clonotypes from each quartile (Q). The score distribution of all cells of the expanded Tex clones is overlaid. (D) Mean Z-scored RNA expression for the pseudo-bulked expanded Tex clonotypes of each quartile for selected general Tex genes and Tex-KLR marker genes. The sign and significance of the Spearman correlation between the avidity-specific gene set module score and each gene are indicated in the quartile 1 (Q1) column. (E) RNA expression for the pseudo-bulked expanded Tex clonotypes following variance stabilized transformation (VST). The large points represent the mean expression for the quartile, and smaller points represent individual clonotypes (n = 82 clonotypes for Q1, 81 each for the other quartiles; DESeq2 Wald test). (F) Expression of the top avidity-specific gene set ( Methods ) and the Tex-KLR signature in FACS-sorted CD8+ TIL subsets from human melanoma samples as quantified by UCell (n = 364 clonotypes from 5 patients for singlet vs. cancer cell doublet and n = 137 clonotypes from 3 patients for singlet vs. APC doublet). Statistical significance was tested using linear regression with patient, clonotype, and log10(clonotype cell count) as fixed-effect covariates.

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: RNA Sequencing, RNA Expression, Clone Assay, Marker, Transformation Assay, Expressing, Cell Characterization

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A–D) Smoothed histograms showing the normalized cell density among all expanded Tex clonotypes for each quartile ( solid line ) with the inter-quartile range ( ribbon ). Data are displayed for the expanded Tex clonotypes in our HNSCC CD8+ TIL dataset (A, B) and the treatment-naïve samples in the CD8 T cell atlas dataset (C, D) , organized by the IL-12-specific gene set ( A, C ) and shared gene set ( B, D ) module scores. Only the genes that were present in > 5% of the cells in at least one cluster were used to apply the module score. (E–H) Heatmaps of the mean Z-scored average RNA expression for the pseudo-bulked Tex clonotypes of each quartile. The sign and significance of the Spearman correlation between the respective module score and each gene are indicated in the quartile 1 (Q1) column. Panels display our HNSCC CD8+ TIL dataset ( E, F ) and CD8 T cell atlas dataset ( G, H ) for the IL-12-specific gene set ( E, G ) and shared gene set ( F, H ) module scores.

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: RNA Expression

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) UMAP plot of the CD8 T cell atlas (Xue et al., 2024). (B) Feature plots showing module scores for each bulk RNA-seq gene set. Only the genes that were present in > 5% of the cells in at least one cluster were used to apply the module score. (C) Violin plots of the module score for each bulk RNA-seq gene set. Datapoints indicate the median module score for each sample in the cluster. (D) Avidity-specific gene set module score of the cells within representative expanded Tex clonotypes from each quartile. The distribution of all cells of expanded Tex clones is shown as an overlaid black line. (E) Smoothed histogram of the normalized avidity-specific gene set module score distributions for the cells in the expanded Tex clonotypes ( solid line ) with the inter-quartile range ( shaded ribbon ). (F) Heatmap of the mean Z-scored RNA expression for the pseudo-bulked expanded Tex clonotypes of each quartile for selected general Tex genes and Tex-KLR marker genes. The sign and significance of the Spearman correlation between the avidity-specific gene set module score with each gene are indicated in the quartile 1 (Q1) column. (G) Pseudo-bulked RNA expression of expanded Tex clonotypes following variance stabilized transformation (VST). The large points connected by the line represent the mean expression, and small points represent individual clonotypes (n = 80 clonotypes for Q1-Q3, 79 for Q4; DESeq2 Wald test).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: RNA Sequencing, Clone Assay, RNA Expression, Marker, Transformation Assay, Expressing

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) UMAP plot of an scRNA-seq dataset of murine CD8 T cells collected during the acute and chronic lymphocytic choriomeningitis virus (LCMV) infection models from the spleen, lung, and liver (Daniel et al., 2022). (B) Feature plots of the module score for each bulk RNA-seq gene set. Genes were converted to the equivalent mouse gene, and only the genes within the gene set that were present in > 5% of the cells in at least one cluster were used to apply the module score. (C-E) Module scores of the (C) avidity-specific, (D) IL-12-specific, and (E) shared gene sets in the CD8 T cell clusters from day 21 post-infection with the chronic LCMV strain. (F) Spearman correlation of Klr genes in the dataset with the avidity-specific gene set module score at day 21 post-infection with the chronic LCMV strain. Bars are colored by -log 10 (FDR). (G) Dot plot of selected activation-associated genes among the Tex clusters from day 21 post-infection with the chronic LCMV model.

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Virus, Infection, RNA Sequencing, Activation Assay

Journal: bioRxiv

Article Title: High-avidity TCR signaling induces a distinct KLR-positive exhaustion state in human tumor-infiltrating CD8 T cells associated with immunotherapy response

doi: 10.64898/2026.05.29.728765

Figure Lengend Snippet: (A) Schematic of clinical trial design. Pathologic treatment response was assessed on the surgical specimen. (B) Percent of Tex-Conv ( left , NKG2A-) and Tex-KLR ( right , NKG2A+) cells of all cells within the peri-tumoral region (200 µm into the stromal region from the tumor-stroma interface) and intra-tumoral region in pre-treatment Cohort 1 samples of the tissue microarray (TMA). A Wilcoxon signed-rank test was used in the comparison between the peri-tumoral and intra-tumoral cell densities within each the non-responders (NR) and responders (R; n = 15 NR and 9 R). A Wilcoxon rank-sum test was used in the comparisons between non-responders and responders in the intra-tumoral region (n = 15 NR and 11 R) and peri-tumoral region (n = 16 NR and 9 R). The p values were not adjusted because the comparisons were pre-determined. (C) Representative images of the TMA IF data of Cohort 1 pre-treatment samples. Selected Tex-KLR cells are indicated with a white arrow. (D) Representative images of the whole tissue section IF data of patient OC36 of Cohort 2 at the pre- and post-treatment timepoints. Selected Tex-KLR cells are indicated with a white arrow. (E-G) Percent of each cell type of total cells within the indicated region at the pre- and post-treatment timepoints for (E) Tex-Conv, (F) Tex-KLR, and (G) Non-Tex CD8 T cells (n = 2 non-responders and 2 responders).

Article Snippet: CD8+ TILs were isolated with the REAlease CD8 (TIL) MicroBead Kit (Miltenyi Biotec, 130-121-560) according to the manufacturer’s protocol.

Techniques: Microarray, Comparison

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting KDM4C enhances CD8 + T cell mediated antitumor immunity by activating chemokine CXCL10 transcription in lung cancer

doi: 10.1136/jitc-2021-003716

Figure Lengend Snippet: KDM4C inhibition enhances CD8 + T cell infiltration and migration. (A) C57BL/6 mice were injected subcutaneously with 1×10 6 stable Lewis cells (sh-control or sh-KDM4C). The volumes of the subcutaneously transplanted tumors were measured every 3 days, and the growth curves were drawn (mean±SEM; n=10; ***p<0.001). (B) All subcutaneously transplanted tumor weights were measured (n=10/group, ***p<0.001). (C) Mice bearing Lewis tumors were treated with SD70 (10 mg/kg) when the tumor volume reached a calculated average of 100 mm 3 . After administration for five consecutive days, SD70 was administered once every 3 days (mean±SEM; n=8; ***p<0.001). (D) All subcutaneously transplanted tumor weights of each group (n=8/group, ***p<0.001). (E, F) The percentages of CD8 + T cells in CD3 + cells in tumor tissues after the indicated treatment. The mean±SD is shown. **P<0.01, ***p<0.001. (G, H) Left panel: representative immunohistochemical staining images of CD8 + T cells in CD3 + cells in tumor tissues after the indicated treatment. Scale bar: 100 µm. Right panel: CD8 + T cells counted with ImageJ software. Three fields were selected for each sample. ***p<0.001. (I, J) The percentages of proliferating CD8 + T cells were analyzed by Ki67 staining. The mean±SD is shown. (K, L) At 72 hours after CFSE staining, the proliferation of CD8 + T cells was measured by flow cytometry. ***P<0.001. (M) Schematic diagram of in vitro CD8 + T cell migration assays. (N, O) The number of CD8 + T cells passing through the membrane of a Transwell system was calculated by flow cytometry. The mean±SD is shown. ***P<0.001.

Article Snippet: Each 100 μL cell suspension was incubated with 10 μL Miltenyi CD8 (TIL) MicroBeads in the dark at 4°C for 15 min, MACS Buffer (130-091-222, Miltenyi Biotec) was added twice for washing, and the cells were resuspended in 1 mL buffer.

Techniques: Inhibition, Migration, Injection, Control, Immunohistochemical staining, Staining, Software, Flow Cytometry, In Vitro, Membrane

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting KDM4C enhances CD8 + T cell mediated antitumor immunity by activating chemokine CXCL10 transcription in lung cancer

doi: 10.1136/jitc-2021-003716

Figure Lengend Snippet: KDM4C inhibition augments antitumor immunity and delays exhaustion in CD8 + T cells in vitro and in vivo. (A–C) CD8 + T cells isolated from tumor tissues after the indicated treatment were stimulated with PMA (100 ng/mL), monensin sodium salt (1 µg/mL), and ionomycin (100 ng/mL) for 6 hours, and cell cytotoxicity markers (IFN-γ, GZMB and Perforin) were assessed by flow cytometry. *P<0.05, ***p<0.001. (D–G) Evaluation of the effect of KDM4C silencing on the cytotoxicity of CD8 + T cells in an in vitro conditioned culture model. ***P<0.001. (H) Specific lysis of Lewis tumor cells by CD8 + T cells pretreated with the indicated conditioned medium. Each experiment was repeated three times independently. ***P<0.001. (I) Lewis cells were cocultured with pretreated CD8 + T cells at the indicated proportion for 24 hours, and the T cells were washed and removed with phosphate-buffered saline, fixed with 4% paraformaldehyde and stained with crystal violet. Each experiment was performed in triplicate. (J) Lewis cells and pretreated CD8 + T cells were cocultured at 1:3. After 24 hours, the tumor cells were stained for an apoptosis assay, and the proportion of apoptotic Lewis cells was detected by flow cytometry. (K, L) After the indicated treatment, the levels of the exhaustion markers PD-1 and CD39 on CD8 + T cells in tumor tissues were assessed. **P<0.01, ***p<0.001. (M, N) Flow cytometry analyses for exhaustion markers on CD8 + T cells treated with the indicated conditioned medium are shown. **P<0.01, ***p<0.001.

Article Snippet: Each 100 μL cell suspension was incubated with 10 μL Miltenyi CD8 (TIL) MicroBeads in the dark at 4°C for 15 min, MACS Buffer (130-091-222, Miltenyi Biotec) was added twice for washing, and the cells were resuspended in 1 mL buffer.

Techniques: Inhibition, In Vitro, In Vivo, Isolation, Flow Cytometry, Lysis, Saline, Staining, Apoptosis Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting KDM4C enhances CD8 + T cell mediated antitumor immunity by activating chemokine CXCL10 transcription in lung cancer

doi: 10.1136/jitc-2021-003716

Figure Lengend Snippet: Enhanced antitumor immunity caused by KDM4C inhibition is mediated by CXCL10. (A) C57BL/6 mice were injected subcutaneously with 1×10 6 Lewis cells. When the tumor volume reached an average of 100 mm 3 , the mice were given a vehicle either alone or in combination with SD70 and/or anti-CXCL10. Tumor volume was measured every 3 days (mean±SEM; n=8; ***p<0.001; two-tailed t-test). (B) Tumor wet weights at 22 days’ postinoculation. n=8, **p<0.01. (C, D) Flow cytometry analyses showing the changes in the percentages of CD8 + T cells (C) and IFN-γ + CD8 + T cells (D) in each group. n=8/group, *p<0.05, ***p<0.001. (E) Changes in Ki67 + CD8 + T cells with treatment. n=3, **p<0.01, ***p<0.001. (F) CFSE staining of proliferating CD8 + T cells measured by flow cytometry. Each experiment was repeated three times independently. (G) Changes in the in vitro migration of CD8 + T cells. n=3, ***p<0.001. (H) Schematic illustration of the in vivo migration experiment. (I, J) IHC was used to analyze the infiltration of CD8 + T cells in subcutaneously transplanted tumors for each group. Scale bar: 100 µm; n=8/group; ***p<0.001. (K, L) Flow cytometry was used to detect the expression of cytotoxicity markers (IFN-γ, GZMB, Perforin, and CD107a) and exhaustion markers (PD-1 and CD39) in CD8 + T cells in each group. n=3; *p<0.05, **p<0.01, ***p<0.001. (M) Lewis cells in 24-well plates were cocultured with pretreated CD8 + T cells in the absence or presence of an anti-CXCL10 neutralizing antibody or control antibody for 24 hours. Crystal violet staining identified the surviving tumor cells. Each group of experiments was carried out three times.

Article Snippet: Each 100 μL cell suspension was incubated with 10 μL Miltenyi CD8 (TIL) MicroBeads in the dark at 4°C for 15 min, MACS Buffer (130-091-222, Miltenyi Biotec) was added twice for washing, and the cells were resuspended in 1 mL buffer.

Techniques: Inhibition, Injection, Two Tailed Test, Flow Cytometry, Staining, In Vitro, Migration, In Vivo, Expressing, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting KDM4C enhances CD8 + T cell mediated antitumor immunity by activating chemokine CXCL10 transcription in lung cancer

doi: 10.1136/jitc-2021-003716

Figure Lengend Snippet: Triple therapy shows the greatest antitumor effects in lung cancer. (A) Schematic illustration of treatment schedule. (B) A C57BL/6 murine model was established, and the mice were randomly divided into eight groups. Each group was treated according to the schedule shown in (A). All subcutaneously transplanted tumors were measured every 3 days with Vernier calipers. When the subcutaneously transplanted tumors in the group reached 1500 mm 3 or more than 60 days had elapsed from tumor cell injection, the experiment was terminated. The tumor growth curves of subcutaneously transplanted tumors were drawn, and the data on the 19th day were statistically analyzed (mean±SEM, n=8/group). (C) Kaplan–Meier analysis across the treatment groups in figure part B. (D, E) Flow cytometry analyses of CD8 + T cells (D) and IFN-γ + CD8 + T cells (E) in subcutaneously transplanted tumors on day 19 of treatment. P<0.05 was considered statistically significant (n=8/group; two-tailed t-test).

Article Snippet: Each 100 μL cell suspension was incubated with 10 μL Miltenyi CD8 (TIL) MicroBeads in the dark at 4°C for 15 min, MACS Buffer (130-091-222, Miltenyi Biotec) was added twice for washing, and the cells were resuspended in 1 mL buffer.

Techniques: Injection, Flow Cytometry, Two Tailed Test

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting KDM4C enhances CD8 + T cell mediated antitumor immunity by activating chemokine CXCL10 transcription in lung cancer

doi: 10.1136/jitc-2021-003716

Figure Lengend Snippet: CD8 + T cells and CXCL10 are necessary for antitumor immunity induced by the triple therapy. (A) Schematic illustration of neutralization assays. Mice received one of five treatments on the indicated days: vehicle, triple therapy, triple therapy with an anti-CD8 neutralizing antibody, triple therapy with an anti-CXCL10 neutralizing antibody, and/or triple therapy with both the anti-CD8 and anti-CXCL10 neutralizing antibodies. (B) Mice (n=8/group) were inoculated with Lewis cells and given treatments as described in figure part A. The volumes of the subcutaneously transplanted tumors were recorded every 3 days, and the data on the 25th day were statistically analyzed (mean±SEM); p<0.05 was considered statistically significant). (C) Kaplan-Meier survival curves of mice bearing tumors in different treatment groups (n=8/group, Gehan-Breslow-Wilcoxon test).

Article Snippet: Each 100 μL cell suspension was incubated with 10 μL Miltenyi CD8 (TIL) MicroBeads in the dark at 4°C for 15 min, MACS Buffer (130-091-222, Miltenyi Biotec) was added twice for washing, and the cells were resuspended in 1 mL buffer.

Techniques: Neutralization

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting KDM4C enhances CD8 + T cell mediated antitumor immunity by activating chemokine CXCL10 transcription in lung cancer

doi: 10.1136/jitc-2021-003716

Figure Lengend Snippet: Schematic diagram showing the mechanism by which KDM4C is involved in antitumor immunity in lung cancer. Left panel: KDM4C is abnormally upregulated in lung cancer cells. By reducing the enrichment of the activated histone H3K36me3 at the promoter of CXCL10 and inhibiting CXCL10 expression, KDM4C reduces the infiltration and activation of CD8 + T cells, which converts the tumor to a ‘cold’ state that is insensitive to radiotherapy and immunotherapy. Right panel: targeted pharmacological inhibition of KDM4C using SD70 can reprogram the epigenetic state in a tumor, activate the expression of CXCL10, and enhance the recruitment and activation of CD8 + T cells, converting the tumor to a ‘hot’ state that is sensitive to radiotherapy and immunotherapy.

Article Snippet: Each 100 μL cell suspension was incubated with 10 μL Miltenyi CD8 (TIL) MicroBeads in the dark at 4°C for 15 min, MACS Buffer (130-091-222, Miltenyi Biotec) was added twice for washing, and the cells were resuspended in 1 mL buffer.

Techniques: Expressing, Activation Assay, Inhibition

Journal: Nature immunology

Article Title: Microbial ligand-independent regulation of lymphopoiesis by NOD1

doi: 10.1038/s41590-023-01668-x

Figure Lengend Snippet: a , Representative contour plots of total (B220 − , NK1.1 − , CD11b − ) thymocytes stained with anti-CD4 and anti-CD8 antibodies (top), DN thymocytes stained with anti-CD44 and anti-CD25 antibodies (middle) and CD44 + DN1 thymocytes stained with anti-ckit and CD25 antibodies (bottom). b , c , Flow cytometry analysis of PDCA-1 and CD8α splenic CD11c + MHC class II + dendritic cells ( b ) and CD62L plus CD44 on splenic CD4 + and CD8 + T lymphocytes ( c ). In each panel, the bars depict numbers (mean ± s.e.m.) for the indicated cell populations per genotype and symbols represent values from individual mice ( a ( n = 5–8), b ( n = 8–9), c ( n = 5–10)) pooled from two independent experiments. Significant differences between groups were determined by two-sided Mann–Whitney test in a and b , and by one-way ANOVA (Tukey’s multiple comparisons) in c and are indicated by asterisks: * P < 0.05, ** P < 0.01, *** P < 0.001. SP, single-positive; DP, double-positive; DN, double-negative thymocytes; ETP, early thymic progenitors; pDC, plasmacytoid dendritic cells; DC, dendritic cells.

Article Snippet: Total CD4 + or CD8 + T lymphocytes (1 × 10 7 ) purified with CD4 or CD8 Micro beads (Miltenyi Biotec, 130–117-043 or 130–117-044) were stimulated with IL-7 (10 ng ml −1 ) or IL-15 (10 ng ml −1 ) for 30 min and then collected.

Techniques: Staining, Flow Cytometry, MANN-WHITNEY

Journal: Nature immunology

Article Title: Microbial ligand-independent regulation of lymphopoiesis by NOD1

doi: 10.1038/s41590-023-01668-x

Figure Lengend Snippet: a , Schematic diagram of breeding strategy used to generate mice with only one WT or one WT and one ΔCARD NOD1 allele. Both crosses were performed in parallel. b , Bars represent the mean ± s.e.m. of cell numbers in indicated populations for WT (n = 9), NOD1 −/− (n = 7), WT ΔCARD NOD1 heterozygote (n = 5) and ΔCARD NOD1 (n = 3) mice. Significant differences between groups were determined by two-sided Mann-Whitney test and indicated by asterisks. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Total CD4 + or CD8 + T lymphocytes (1 × 10 7 ) purified with CD4 or CD8 Micro beads (Miltenyi Biotec, 130–117-043 or 130–117-044) were stimulated with IL-7 (10 ng ml −1 ) or IL-15 (10 ng ml −1 ) for 30 min and then collected.

Techniques: Comparison, MANN-WHITNEY

Journal: Nature immunology

Article Title: Microbial ligand-independent regulation of lymphopoiesis by NOD1

doi: 10.1038/s41590-023-01668-x

Figure Lengend Snippet: Number of splenocytes and indicated lymphocyte populations in spleens from WT, NOD1 −/− or ΔCARD NOD1 control and mice implanted with mini-osmotic pumps releasing rIL-15 for two weeks. The analysis was performed one week after the treatment was stopped. Each symbol represents a value obtained for an individual mouse and bars indicate the mean ± s.e.m. in untreated (n = 7–11) or treated (n = 7–8) group pooled from two independently performed experiments. Significant differences between the groups were determined by two-sided Mann-Whitney test and indicated by asterisks * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Total CD4 + or CD8 + T lymphocytes (1 × 10 7 ) purified with CD4 or CD8 Micro beads (Miltenyi Biotec, 130–117-043 or 130–117-044) were stimulated with IL-7 (10 ng ml −1 ) or IL-15 (10 ng ml −1 ) for 30 min and then collected.

Techniques: In Vivo, Control, MANN-WHITNEY

Journal: Nature immunology

Article Title: Microbial ligand-independent regulation of lymphopoiesis by NOD1

doi: 10.1038/s41590-023-01668-x

Figure Lengend Snippet: Naïve CD8 + T lymphocytes from WT, NOD1 −/− and ΔCARD NOD1 mice were pretreated with or without rIL-15 (10 ng/ml) and cytosolic and nuclear fractions were probed with anti-STAT5 and anti-pSTAT5 Ab. Numbers indicate the intensity of STAT5 bands compared to tubulin or histone H3 protein controls for each cell compartment. Data shown from one representative out of two experiments performed.

Article Snippet: Total CD4 + or CD8 + T lymphocytes (1 × 10 7 ) purified with CD4 or CD8 Micro beads (Miltenyi Biotec, 130–117-043 or 130–117-044) were stimulated with IL-7 (10 ng ml −1 ) or IL-15 (10 ng ml −1 ) for 30 min and then collected.

Techniques: Phospho-proteomics, Translocation Assay

Journal: Molecular Therapy

Article Title: Immune Modulation by Telomerase-Specific Oncolytic Adenovirus Synergistically Enhances Antitumor Efficacy with Anti-PD1 Antibody

doi: 10.1016/j.ymthe.2020.01.003

Figure Lengend Snippet: Recruitment of CD8-Positive Lymphocytes to Tumors and Development of Acquired Antitumor Immunity Mediated by OBP-502

Article Snippet: After CD8-positive selection by CD8 (TIL) MicroBeads (Miltenyi Biotec) and LS Columns (Miltenyi Biotec), Colon26-GFP cells (target) were incubated with the CD8-positive cells (Effector) for 4 h at 37°C at different effector/target ratios (0:1, 10:1, and 20:1).

Techniques: