Journal: iScience

Article Title: Transient tumor exposure induces persistent functional defects in memory CD8 + T cells

doi: 10.1016/j.isci.2026.115556

Figure Lengend Snippet: Phenotypic and transcriptional differences of memory CD8 + T cells generated after a viral or a tumoral challenge Naive F5 x CD45.1 cells (2.10 5 ) were i.v. transferred in B6 mice 1-day prior immunization with VV-NP68 (i.n., 2.10 5 pfu) or EL4-NP68 cells (s.c., 2,5.10 6 cells). (A) Viral load was measured in the lung by qPCR, or tumor volume (mm 3 ) was assessed by measuring its length, width, and thickness over time. (B) The number of Vir-CD8 + and Tum-CD8 + cells was determined over time in the blood by flow cytometry. (C and D) The expression of Ki67 (C) and Bcl2 (D) by Vir-CD8 + and Tum-CD8 + cells was measured over time in the blood. (E) The phenotype of Vir-CD8 + and Tum-CD8 + cells was analyzed 31 days after infection in the spleen, and the percentages of cells expressing each marker are represented as a heatmap. The statistical significance of differences was determined using a two-way ANOVA (C–E) (∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001). Data are represented as mean ± SD and are representative of 3 independent experiments ( n = 5 to 10 mice per group). (F–I) 60 days after immunization, naive F5 and Vir-CD8 + and Tum-CD8 + cells were single-cell sorted, and stimulated with NP68 peptide (10 nM) for 2 h or left untreated. The transcriptome was analyzed by scRNAseq ( n = 476 cells). (F) Clustering of cells projected on a UMAP colored by populations. (G) Proportion of sorted populations in each cluster. (H and I) Volcano plot of the differentially expressed genes between quiescent (H) or restimulated (I) Vir-CD8 + and Tum-CD8 + .

Article Snippet: EL4-NP68 , Dr. T.N.M. Schumacher , Modified from EL4 lymphoma cell line (ATCC “TIB-39”).

Techniques: Generated, Flow Cytometry, Expressing, Infection, Marker, Single Cell

Journal: iScience

Article Title: Transient tumor exposure induces persistent functional defects in memory CD8 + T cells

doi: 10.1016/j.isci.2026.115556

Figure Lengend Snippet: Tum-CD8 + memory cells express molecules associated with T cell exhaustion Naive F5 x CD45.1 cells (2.10 5 ) were i.v. transferred in B6 mice 1-day prior immunization with VV-NP68 (i.n., 2.10 5 pfu) or EL4-NP68 cells (s.c., 2,5.10 6 cells). (A and B) The expression of PD-1, TIM-3, CD9, and Gal3 was measured at the surface of CD8 + memory cells at 30 dpi by flow cytometry. (C and D) The expression of Gal3 was measured intracellularly in memory CD8 + T cells at 30 dpi by flow cytometry. (E) At 30 dpi, splenocytes were stimulated with NP68 peptide (10 nM) for 4 h, and the expression of PD-1, TIM-3, or intracellular Gal3 by memory CD8 + T cells was measured by flow cytometry. The statistical significance of differences was determined using the Mann-Whitney test (B and D) or two-way ANOVA (E) (∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001). Data are represented as mean ± SD ( n = 5 mice per group) and are representative of 3 independent experiments.

Article Snippet: EL4-NP68 , Dr. T.N.M. Schumacher , Modified from EL4 lymphoma cell line (ATCC “TIB-39”).

Techniques: Expressing, Flow Cytometry, MANN-WHITNEY

Journal: iScience

Article Title: Transient tumor exposure induces persistent functional defects in memory CD8 + T cells

doi: 10.1016/j.isci.2026.115556

Figure Lengend Snippet: Tum-CD8 + memory cells display altered cytokine production but not cytotoxic capacities compared to Vir-CD8 + memory cells Naive F5 x CD45.1 cells (2.10 5 ) were i.v. transferred in B6 mice 1-day prior immunization with VV-NP68 (i.n., 2.10 5 pfu) or EL4-NP68 cells (s.c., 2,5.10 6 cells). (A and B) At 30 dpi, F5 memory cells were restimulated with NP68 (10 nM) for 4h in the presence of GolgiStop. The production of IFNγ, TNF, and IL-2 was measured by flow cytometry and expressed in % of total F5 (A) or MFI within the cytokine+ cells (B). (C and D) At 30 dpi, F5 memory cells were restimulated with NP68 (10 nM) for 4h in the presence (cytokines) or absence (CD69) of GolgiStop. The production of IFNγ and TNF and the upregulation of CD69 were measured by flow cytometry over time and expressed in % (C) or MFI (D). (E) The production of IFNγ was measured in supernatant after 4 or 24h of stimulation. (F and G) At 30 dpi, F5 memory cells were restimulated with various doses of NP68 for 4h in the presence of GolgiStop, and the production of IFNγ and TNF was measured by flow cytometry (F). EC 50 was determined (G). (H and I) Splenocytes were incubated with NP68 (10 nM) or control medium for 2h and labeled with CTV or CFSE, respectively. A 1:1 ratio of NP68-loaded splenocytes: control splenocytes (2.10 6 cells) was injected i.v. in Tum-CD8 + or Vir-CD8 + challenged mice at the memory stage. Representative histograms depicting control and CTV-labeled NP68-loaded splenocytes are shown (H). The percentage of NP68-loaded splenocytes killed was evaluated at 6-, 16-, or 44-h post-transfer (I). (J)Total CD8 + enriched from Tum-CD8 + or Vir-CD8 + challenged mice were labeled with CTV and stimulated with NP68-loaded DCs (1:1 ratio) for 4 days in the presence of IL-2. The expansion index of F5 cells was determined after 4 days. The statistical significance of differences was determined using 2-way ANOVA (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗ p < 0.0001). Data are represented as mean ± SD ( n = 5 mice per group) and are representative of 3 independent (A–G) or 1 (H–J) experiment(s).

Article Snippet: EL4-NP68 , Dr. T.N.M. Schumacher , Modified from EL4 lymphoma cell line (ATCC “TIB-39”).

Techniques: Flow Cytometry, Incubation, Control, Labeling, Injection

Journal: iScience

Article Title: Transient tumor exposure induces persistent functional defects in memory CD8 + T cells

doi: 10.1016/j.isci.2026.115556

Figure Lengend Snippet: A transient tumoral challenge is sufficient to alter the protection capacity of F5 memory cells Naive F5 x CD45.1 cells (2.10 5 ) were i.v. transferred in B6 mice 1-day prior immunization with VV-NP68 (i.n., 2.10 5 pfu) or EL4-NP68 cells (s.c., 2,5.10 6 cells). (A–D) At 30 dpi, Vir- or Tum-challenged mice were infected with VV-NP68 (2.10 5 pfu). Six days after infection, mice received an i.v. injection of anti-CD8 antibody, and the proportion of cells within the tissue and the vasculature of the lung was determined (A). The proportion of memory CD8 + T cells in the lung tissue among all memory CD8 + T cells was determined (B). The expression of CD49a (C) and CD49d (D) was measured on memory CD8 + T cells within the lung tissue and vasculature. (E) At 30 dpi, Vir- or Tum-challenged mice were infected with Flu-NP68 (5.10 4 TCID50), and the weight loss was followed for 6 days. (F) At 30 dpi, Vir-CD8 + and Tum-CD8 + memory cells were FACS-sorted and transferred into B6 host (1,2.10 5 cells, i.v. route). One day after transfer, mice received a lethal dose of Flu-NP68 (2.10 6 TCID 50), and survival was followed for 10 days. The statistical significance of differences was determined with 1-way (B) or 2-way (C–E) ANOVA test (∗ p > 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001), or log rank test (F). Data are represented as mean ± SD ( n = 5 mice per group) and are representative of 2 (A–D, and G) or 1 (E and F) experiment(s).

Article Snippet: EL4-NP68 , Dr. T.N.M. Schumacher , Modified from EL4 lymphoma cell line (ATCC “TIB-39”).

Techniques: Infection, Injection, Expressing

Journal: iScience

Article Title: Transient tumor exposure induces persistent functional defects in memory CD8 + T cells

doi: 10.1016/j.isci.2026.115556

Figure Lengend Snippet: Phenotype and cytokine production capacity of F5 memory cells is conserved after homologous or heterologous recall (A) Naive F5 x CD45.1 cells (2.10 5 ) were i.v. transferred in B6 mice 1-day prior immunization with VV-NP68 (i.n., 2.10 5 pfu) or EL4-NP68 cells (s.c., 2,5.10 6 cells). At 26 dpi, mice received a second immunization with VV-NP68 or EL4-NP68. (B) Thirty-one days post-recall, the number of F5 cells was measured in the spleen. (C) The expression of CD9, CD43, CD49a, and CD49d was measured on F5 memory cells 31 days after recall by flow cytometry. (D and E) Splenocytes were stimulated with NP68 (10 nM) for 4h in the presence (D) or absence (E) of GolgiStop. (D) The production of IFNγ, TFNα, and IL-2 was measured by flow cytometry. (E) The expression of PD-1 and TIM3 on F5 memory cells was determined in the spleen. The statistical significance of differences was determined with 1-way (B and C) or 2-way (D and E) ANOVA test (∗ p > 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). Data are represented as mean ± SD ( n = 5 mice per group) and are representative of 2 independent experiments.

Article Snippet: EL4-NP68 , Dr. T.N.M. Schumacher , Modified from EL4 lymphoma cell line (ATCC “TIB-39”).

Techniques: Expressing, Flow Cytometry

Journal: Blood Advances

Article Title: Serum free fatty acid promotes tumor progression and predicts golidocitinib sensitivity in peripheral T-cell lymphoma ∗

doi: 10.1182/bloodadvances.2025017682

Figure Lengend Snippet: High serum FFA correlated with tumor JAK-STAT signaling activation. (A) Pathways enriched with differentially expressed genes from the FFA-high group compared with the FFA-low group in all patients with PTCL, NKTCL, nTFHL, and PTCL-NOS. (B) Correlations between indicated lipid metabolism pathways and the JAK-STAT signaling pathway in all patients with PTCL, NKTCL, nTFHL, and PTCL-NOS. (C) Schema of C57BL/6 syngeneic murine model of EL4 cells in mice fed with CD or HFD. (D) The FFA levels in plasma (left) and tumor tissues (right) derived from implanted tumor model fed with CD and HFD. (E) Representative images (left) and quantification of Oil Red O (ORO) staining (right) in tumor tissues derived from implanted tumor model fed with CD and HFD. (F) Tumor volumes of EL4 subcutaneous tumor model fed with CD and HFD. (G) Protein expression of JAK1, STAT3, and p-STAT3 (Tyr705) in tumor tissues derived from implanted tumor model fed with CD and HFD. (H) Representative images (upper) and statistical analysis (lower) of p-STAT3 (Tyr705) positivity in tumor tissues derived from FFA-high and -low patients of NKTCL, nTFHL, and PTCL-NOS, respectively. P values in panel A are obtained through enrichment analysis, P values in panel B are calculated with Spearman’ rank analysis, P values in panels D-F,H are calculated with a 2-tailed Student t test. Data are presented as mean ± standard error of the mean (SEM).

Article Snippet: THP-1 (a human acute monocytic leukemia), Karpas299, Jurkat, and EL4 cell lines were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Activation Assay, Clinical Proteomics, Derivative Assay, Staining, Expressing

Journal: Blood Advances

Article Title: Serum free fatty acid promotes tumor progression and predicts golidocitinib sensitivity in peripheral T-cell lymphoma ∗

doi: 10.1182/bloodadvances.2025017682

Figure Lengend Snippet: Golidocitinib showed therapeutic potential via modulating suppressive TME. (A) Schema of C57BL/6 syngeneic murine model of EL4 cells with therapeutic intervention. (B) Tumor volumes of C57BL/6 syngeneic murine model of EL4 cells fed with CD and HFD upon treatment with golidocitinib or vehicle. (C) Relative IL6 (left) and IL10 (right) expression of C57BL/6 syngeneic murine model of EL4 cells fed with CD and HFD upon treatment with golidocitinib or vehicle. (D) Representative images (left) and statistical analysis (right) of the proportions of MDSCs and M2 macrophages in tumor tissues derived from C57BL/6 syngeneic murine model of EL4 cells fed with CD and HFD upon treatment with golidocitinib or vehicle. (E) Representative images (upper) and statistical analysis (lower) of mean fluorescence intensity (MFI) of CD44 on CD8 + T cells in tumor tissues derived from implanted tumor model fed with CD and HFD upon treatment with golidocitinib or vehicle. (F) Representative images (left) and statistical analysis (right) of STAT3 and p-STAT3 positivity in tumor tissues derived from implanted tumor model fed with CD and HFD upon treatment with golidocitinib or vehicle. P values in panels B-F are calculated with a 2-tailed Student t test. Data are presented as mean ± SEM.

Article Snippet: THP-1 (a human acute monocytic leukemia), Karpas299, Jurkat, and EL4 cell lines were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Derivative Assay, Fluorescence

Journal: Cellular Oncology

Article Title: BCAA catabolism mediates POU2AF1 propionylation to enhance T-ALL development

doi: 10.1007/s13402-026-01201-w

Figure Lengend Snippet: POU2AF1 promotes T-ALL development via transcriptional activation of SLC7A11. ( A ) Western blot analysis of SLC7A11 protein levels in the EL4 cell lines. ( B ) The protein level of SLC7A11 was determined in WT and KO T-ALL cells by Western blotting ( n = 3). ( C ) POU2AF1 and SLC7A11 levels were measured in bone marrow cells from transplanted WT, Bcat1 -KO, Pou2af1 -overexpressing WT or Bcat1 -KO mice by Western blotting. ( D ) Luciferase reporter assay evaluating the transcriptional activation of Slc7a11 by Pou2af1 -WT and Pou2af1 -K5R ( n = 3). ( E ) ChIP assay showing the binding of Pou2af1 -WT and Pou2af1 -K5R to the Slc7a11 promoter region ( n = 3). ( F – G ) Representative flow cytometric analysis of GFP + mCherry + leukemia cells (The markers for the indication of leukemia cells were GFP + and mCherry + , which were the tags for the MSCV- Notch1 -IRES-GFP plasmid and MSCV-mCherry overexpression plasmid) in peripheral blood from recipient mice 3 weeks after transplantation with WT and KO T-ALL cells overexpressing Slc7a11 . ( H – I ) Representative images and quantification of the liver, spleen and thymus morphology in recipient mice ( n = 3). ( J ) Survival analysis of recipient mice ( n = 5). The data are presented as the means ± SDs. One-way ANOVA with Tukey’s multiple comparison test ( G ), log-rank test ( J ) and two-way ANOVA with Sidak’s multiple comparison test ( D , I ) were used for comparisons of statistical significance (*, P < 0.05; **, P < 0.01; and ***, P < 0.001)

Article Snippet: These viral particles were subsequently used to infect NOTCH-GFP+ bone marrow leukemia cells, human cell lines Jurkat and MOLT-4 (ATCC), murine cell lines EL4 and L1210 (ATCC), as well as primary T-ALL patient samples, followed by assessment of in vitro cell proliferation.

Techniques: Activation Assay, Western Blot, Luciferase, Reporter Assay, Binding Assay, Plasmid Preparation, Over Expression, Transplantation Assay, Comparison

Journal: Cellular Oncology

Article Title: BCAA catabolism mediates POU2AF1 propionylation to enhance T-ALL development

doi: 10.1007/s13402-026-01201-w

Figure Lengend Snippet: Targeting BCAT1 metabolism significantly impaired T-ALL progression. ( A – D ) Analysis of the proliferation of EL4 ( A ), L1210 ( B ), Jurkat ( C ), and MOLT-4 ( D ) cells treated in vitro with 10 mM gabapentin (n=4). ( E ) Representative flow cytometric analysis of peripheral blood at 3 weeks post-transplantation from T-ALL model mice following treatment with IgG, anti-PD-1 antibody, low-BCAA, or combined low-BCAA + anti-PD-1 antibody. ( F ) Quantification of the peripheral blood data shown in Panel E (n=5). ( G ) Overall survival analysis of treated mice (n=5). ( H ) Schematic model illustrating how BCAT1-driven branched-chain amino acid metabolism promotes T-ALL progression via Kpr-mediated activation of POU2AF1. The data are presented as the means ± SDs. One-way ANOVA with Tukey’s multiple comparison test ( F ), log-rank test ( G ), and two-way ANOVA with Sidak’s multiple comparison test (A–D) were used for comparisons of statistical significance (*, P <0.05; **, P <0.01; and ***, P <0.001)

Article Snippet: These viral particles were subsequently used to infect NOTCH-GFP+ bone marrow leukemia cells, human cell lines Jurkat and MOLT-4 (ATCC), murine cell lines EL4 and L1210 (ATCC), as well as primary T-ALL patient samples, followed by assessment of in vitro cell proliferation.

Techniques: In Vitro, Transplantation Assay, Activation Assay, Comparison

Journal: Cell Reports Medicine

Article Title: Targeting BCMA in multiple myeloma with a trifunctional NK cell engager

doi: 10.1016/j.xcrm.2026.102628

Figure Lengend Snippet: 1:1 BCMA-NKp46-FcγRIIIa NKCE drives anti-tumor activity in vitro and in vivo (A) Schematic of BCMA-NKp46-FcγRIIIa NKCE mode of action. The molecule binds to BCMA on MM cells and recruits/activates NK cells by co-engaging NKp46 and FcγRIIIa, leading to MM cell killing and/or cytokine production. (B) (Left) Cytotoxicity comparison of F25, CODV-1:1, CODV-2:1, BCMA-IgG1, and IC-NKp46-FcγRIIIa NKCE control (IC-F25). RPMI 8226 and MM.1S MM cells were used as targets, with purified resting NK cells from 4 healthy donors as effectors. (Right) EC 50 and maximum cytotoxic activity of BCMA-IgG1 and BCMA-NKp46-FcγRIIIa NKCE against RPMI 8226 and MM.1S MM cells. Delta maximum lysis (ΔMax lysis)—defined as percent maximum lysis minus background lysis of the isotype control (IC-F25) at corresponding concentration—and EC 50s were monitored from dose-response curves after 4-h incubation, plotted separately for all MM cell lines and NK donors ( n = 4). ANOVA/Tukey’s multiple comparisons test ∗ p < 0.05, ∗∗∗ p < 0.0005. (C) Comparison of cytotoxicity between 1:1 (F25, CODV-1:1) and 2:2 (F33, CODV-2:2) BCMA-NKp46-FcγRIIIa NKCE formats with the same setting as in (B). Two representative NK donors are shown ( n = 17). (D) (Left) Flow cytometry analysis of fluorescent RMA cells expressing human BCMA ( dsRed ) or lacking BCMA expression ( eGFP ), showing BCMA expression only on dsRed cells. (Middle) Experimental setup. A 2:3 mixture of eGFP (BCMA-negative) and dsRed (BCMA-positive) RMA cells was intravenously injected into huNKp46-Tg x Rag1 −/− transgenic mice. Tumor-bearing mice ( n = 27–28 per group) received a single dose of 12.5 pmol/animal (0.078 mg/kg) of BCMA-NKp46-FcγRIIIa NKCEs or vehicle control. Liver biopsies were collected 48 h post-treatment, and infiltrating RMA cells quantified by flow cytometry. (Right) Flow cytometry analysis of BCMA expression on dsRed RMA cells pre-engraftment ( in vitro , red histogram) and post-engraftment in liver biopsies (gray histogram). (E) (Left) Absolute count of liver-infiltrated RMA cells. (Right) dsRed/eGFP cell ratio in liver biopsies, analyzed by flow cytometry 48 h after treatment. Statistical analysis: Mann-Whitney test (ns, p > 0.05; ∗ p < 0.01). (F) Kaplan-Meier survival curves for huNKp46-Tg x Rag1−/− mice bearing disseminated EL4-huBCMA tumors treated with BCMA-NKp46-FcγRIIIa NKCEs lacking Fc optimization for enhanced ADCC. Treatment groups (red) received 5, 0.5, or 0.05 mg/kg of BCMA-NKp46-FcγRIIIa NKCE, compared to the isotype control NKCE (gray, 5 mg/kg) and vehicle (black). Statistical analysis: log rank (Mantel-Cox) test ( p < 0.05, ∗ p = 0.0398, ∗∗ p = 0.0082, ∗∗∗ p = 0.0001, ∗∗∗∗ p < 0.0001). See also .

Article Snippet: huBCMA-expressing EL4 cells (wild type EL4 cell line obtained from ATCC, TIB-39) , Sanofi , This paper.

Techniques: Activity Assay, In Vitro, In Vivo, Comparison, Control, Purification, Lysis, Concentration Assay, Incubation, Flow Cytometry, Expressing, Injection, Transgenic Assay, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Targeting BCMA in multiple myeloma with a trifunctional NK cell engager

doi: 10.1016/j.xcrm.2026.102628

Figure Lengend Snippet: ADCC-optimized BCMA-NKp46-FcγRIIIa NKCE drives anti-tumor activity in vitro and in vivo (A) Comparison of cytotoxicity of BCMA-NKp46-FcγRIIIa NKCE molecules engineered for enhanced ADCC (-ADE) vs. non-engineered (wild-type Fc) counterparts. RPMI 8226 MM cells were used as target, and purified resting NK cells served as effector. Data from one representative NK cell donor is shown ( n = 13). (B) EC 50 and maximum cytotoxic activity of F25 ( N = 21), F25-ADE ( N = 13), CODV-1:1 ( N = 29), CODV-1:1-ADE ( n = 14), F33 ( n = 15), F33-ADE ( n = 13), CODV-2:2 ( n = 23), and CODV-2:2-ADE ( n = 13) molecules against RPMI 8226 MM cells. ΔMax lysis and EC 50s were determined from dose-response curves and plotted separately for each NK donor. Statistical analysis: ANOVA/Tukey’s multiple comparisons test (∗ p < 0.005; ∗∗∗ p < 0.0001). (C) Kaplan-Meier survival curves for huFcγR-Tg mice bearing disseminated EL4-huBCMA tumors and treated with surrogate BCMA-moNKp46-FcγRIIIa NKCE lacking ADCC optimization (wild-type Fc) ( CODV-1 : 1 , pink) or ADCC-optimized BCMA-moNKp46-FcγRIIIa NKCE ( CODV-1 : 1-ADE , blue) at 0.5 mg/kg. Statistical analysis: log rank (Mantel-Cox) test (∗∗ p < 0.001). See also and .

Article Snippet: huBCMA-expressing EL4 cells (wild type EL4 cell line obtained from ATCC, TIB-39) , Sanofi , This paper.

Techniques: Activity Assay, In Vitro, In Vivo, Comparison, Purification, Lysis