Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Human CD3 complex is required for the generation of T-cell receptor-like chimeric antigen receptor targeting WT1 in natural killer cells

doi: 10.1007/s00262-026-04352-9

Figure Lengend Snippet: Construction of CAR-WT1 specific for WT1(126–134) peptide presented by HLA-A2. A Schematic diagram of the third-generation CAR-WT1 construct harboring signal peptide, TCR-like scFv, CD8 hinge, CD28, 4-1BB, and CD3ζ, under EF-1α promoter. B Schematic illustration of the generation of CAR-WT1 T cells in primary T cells. C Flow cytometric analysis of surface CAR expression in CAR-WT1 T cells using F(ab’)2 antibody. D Flow cytometric gating strategy to specifically detect cell death in PKH67-labeled target tumor (T) cells in response to effector (E) cells, e.g., T or NK cells, using Annexin V/7-AAD assay. Debris was excluded from the analysis (see also Suppl. Fig. ). E Percentage of total tumor cell death, comprising Annexin V + and/or 7-AAD + cells, of WT1 + /HLA-A2 + Jeko-1 and THP-1 cells, as well as WT1 + /HLA-A2 – K562 and HL60 cells in response to CAR-WT1 or non-transduced (NTD) T cells at various E:T ratios and at the indicated time. Data are mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 versus NTD T cells under the same conditions; two-sided Student’s t -test

Article Snippet: Human NK-92 cells, human Jurkat T (clone E6-1) cells, and human mantle cell lymphoma (MCL) Jeko-1 cells were obtained from American Type Cell Collection (ATCC; Manassas, VA), while other human hematologic cancer cell lines, including THP-1 (JCRB0112), K562 (JCRB0019), and HL60 (JCRB0085) were obtained from Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan).

Techniques: Construct, Expressing, Labeling

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Human CD3 complex is required for the generation of T-cell receptor-like chimeric antigen receptor targeting WT1 in natural killer cells

doi: 10.1007/s00262-026-04352-9

Figure Lengend Snippet: CAR-WT1/CD3 NK-92 cells effectively induce cell death of WT1 + /HLA-A2 + target tumor cells. A Tumor cells, including WT1 + /HLA-A2 + Jeko-1 and THP-1 cells, as well as WT1 + /HLA-A2 – K562 and HL60 cells, were pre-labeled with PKH67 dye and NK cytotoxicity of CAR-WT1/CD3 and NTD NK-92 cells at various E:T ratios was evaluated by Annexin V/7-AAD assay at the indicated time using the same gating strategy shown in Suppl. Fig. . Data are mean ± SD (n = 3 or 5). * P < 0.05, ** P < 0.01, *** P < 0.001 versus NTD NK-92 cells under the same conditions; two-sided Student’s t -test. B The proportion of CAR-WT1/CD3 or NTD NK-92 cells forming binding doublets with Jeko-1 (left) or HL60 (right) cells, expressed as a percentage of all effector cells at various E:T ratios (see Suppl. Fig. for the gating strategy). Negative (Neg) controls were NK-92 and target cells at the E:T ratio of 1:1 incubated at 4 °C. Data are mean ± SD (n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001 versus NTD NK-92 cells under the same conditions; two-sided Student’s t -test

Article Snippet: Human NK-92 cells, human Jurkat T (clone E6-1) cells, and human mantle cell lymphoma (MCL) Jeko-1 cells were obtained from American Type Cell Collection (ATCC; Manassas, VA), while other human hematologic cancer cell lines, including THP-1 (JCRB0112), K562 (JCRB0019), and HL60 (JCRB0085) were obtained from Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan).

Techniques: Labeling, Binding Assay, Incubation

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Human CD3 complex is required for the generation of T-cell receptor-like chimeric antigen receptor targeting WT1 in natural killer cells

doi: 10.1007/s00262-026-04352-9

Figure Lengend Snippet: Activation and function of CAR-WT1/CD3 NK-92 cells in response to target tumor cells. CAR-WT1/CD3 NK-92 cells were exposed with or without WT1 + /HLA-A2 + Jeko-1 cells at the E:T ratio of 1:1 for 4 h. A Flow cytometric analysis of surface CD107a in CD56 + CAR-WT1/CD3 or NTD NK-92 cells (see Suppl. Fig. for the gating strategy and representative plots). Data are mean ± SD (n = 3). * P < 0.05 versus NTD NK-92 cells under the same conditions; two-sided Student’s t -test. B Quantitative measurement of IFN- γ and TNF-⍺ by ELISA in cell-free supernatant. Data are mean ± SD (n = 3). *** P < 0.001 versus NTD NK-92 cells under the same conditions; two-sided Student’s t -test. ND, not detectable. C Jeko-1 cells were pre-labeled with PKH67 dye before co-exposure with CAR-WT1/CD3 or NTD NK-92 cells. (upper) mRNA expression of genes involved in immune cell signaling was evaluated in PKH67 – NK-92 cells by qPCR, normalized to GAPDH , and represented in a heatmap. (lower) mRNA expression of certain genes, e.g., IFNA21 , IFNG , and TNF , are plotted. Data are mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 versus NTD NK-92 cells alone; # P < 0.05, ### P < 0.001 versus the indicated group; one-way ANOVA with Tukey’s post- test. NS, not significant

Article Snippet: Human NK-92 cells, human Jurkat T (clone E6-1) cells, and human mantle cell lymphoma (MCL) Jeko-1 cells were obtained from American Type Cell Collection (ATCC; Manassas, VA), while other human hematologic cancer cell lines, including THP-1 (JCRB0112), K562 (JCRB0019), and HL60 (JCRB0085) were obtained from Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Labeling, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Human CD3 complex is required for the generation of T-cell receptor-like chimeric antigen receptor targeting WT1 in natural killer cells

doi: 10.1007/s00262-026-04352-9

Figure Lengend Snippet: Surface CD3 is not required for the antitumor activity of CAR-WT1/CD3 NK-92 cells. A Gating strategy for FACS sorting of CAR-WT1/CD3 NK-92 cells according to its surface CD3, into: (i) mixed surface CD3 +/– ; (ii) surface CD3 – ; and (iii) surface CD3 + , all with surface CAR-WT1 + . B , C mRNA expression of CAR[ CD3Z ] (B) and CD3D , CD3E , and CD3Z (C) in different subgroups of sorted CAR-WT1 + NK-92 cells by qPCR. Data are mean ± SD (n = 3). *** P < 0.001 versus NTD NK-92 cells; # P < 0.05, ## P < 0.01, ### P < 0.001 versus the indicated group; one-way ANOVA with Tukey’s post-test. NS, not significant. D , E Different subgroups of sorted CAR-WT1 + NK-92 cells were exposed to Jeko-1 cells at the E:T ratio of 1:1 for 4 h. D Percentage of total Jeko-1 tumor cell death, as evaluated by Annexin V/7-AAD assay using the same gating strategy shown in Suppl. Fig. . E Percentage of surface CD107a in CD56 + NK-92 cells (see Suppl. Fig. for the gating strategy). For panels D and E, data are mean ± SD (n = 6). ** P < 0.01, *** P < 0.001 versus NTD NK-92 cells at the E:T ratio of 1:1; one-way ANOVA with Tukey’s post-test. NS, not significant

Article Snippet: Human NK-92 cells, human Jurkat T (clone E6-1) cells, and human mantle cell lymphoma (MCL) Jeko-1 cells were obtained from American Type Cell Collection (ATCC; Manassas, VA), while other human hematologic cancer cell lines, including THP-1 (JCRB0112), K562 (JCRB0019), and HL60 (JCRB0085) were obtained from Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan).

Techniques: Activity Assay, Expressing

Journal: Translational Cancer Research

Article Title: Curcumin inhibits the proliferation of diffuse large B-cell lymphoma by inducing ferroptosis via the ACSL4-SAT1-GPX4 axis

doi: 10.21037/tcr-2025-1-2674

Figure Lengend Snippet: Curcumin exhibits favorable drug-like properties and inhibits DLBCL cell proliferation in a concentration- and time-dependent manner. (A) Prediction of drug-likeness and oral bioavailability for curcumin using the SwissADME database. (B,C) The viability of SU-DHL-2 (B) and OCI-LY7 (C) cells after treatment with increasing concentrations of curcumin for 24, 48, and 72 hours, as determined by the CCK-8 assay. Data are presented as the mean ± standard deviation (n=3). The calculated IC 50 values at each time point are indicated. CCK-8, Cell Counting Kit-8; DLBCL, diffuse large B-cell lymphoma; IC 50 , half maximal inhibitory concentration.

Article Snippet: The human lymphoma cell lines OCI-LY7 (KyeGEN, KGG3250-1) and SU-DHL-2 (KeyGEN, KGG3254-1) were purchased from Jiangsu KeyGEN BioTECH Co., Ltd. (Nanjing, China).

Techniques: Concentration Assay, CCK-8 Assay, Standard Deviation, Cell Counting

Journal: Translational Cancer Research

Article Title: Curcumin inhibits the proliferation of diffuse large B-cell lymphoma by inducing ferroptosis via the ACSL4-SAT1-GPX4 axis

doi: 10.21037/tcr-2025-1-2674

Figure Lengend Snippet: Curcumin treatment enriches the ferroptosis pathway and modulates the expression of key ferroptosis-related proteins. (A) Principal component analysis and correlation analysis showing distinct clustering of samples from curcumin-treated and control OCI-LY7 cells. (B) Volcano plot showing DEGs in OCI-LY7 cells after curcumin treatment compared to the control group. (C) Heatmap indicated the number of upregulated and downregulated DEGs. (D) KEGG pathway enrichment analysis for DEGs. DEGs, differentially expressed genes; KEGG, Kyoto Encyclopedia of Genes and Genomes; OC, OCI-LY7 control; OC_Cur, OCI-LY7 treated with curcumin.

Article Snippet: The human lymphoma cell lines OCI-LY7 (KyeGEN, KGG3250-1) and SU-DHL-2 (KeyGEN, KGG3254-1) were purchased from Jiangsu KeyGEN BioTECH Co., Ltd. (Nanjing, China).

Techniques: Expressing, Control

Journal: Translational Cancer Research

Article Title: Curcumin inhibits the proliferation of diffuse large B-cell lymphoma by inducing ferroptosis via the ACSL4-SAT1-GPX4 axis

doi: 10.21037/tcr-2025-1-2674

Figure Lengend Snippet: Curcumin regulates the expression of key ferroptosis-related proteins in a concentration-dependent manner. (A) Curcumin treatment upregulated the pro-ferroptotic proteins ACSL4 and SAT1, while downregulating the anti-ferroptotic protein GPX4. (B) Western blot was used to detect the protein expression levels of ACSL4, SAT1, and GPX4 in OCI-LY7 cells treated with increasing concentrations of curcumin. Data are presented as the mean ± standard deviation (n=3). ns, P>0.05; *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. OC, OCI-LY7 control; OC_Cur, OCI-LY7 treated with curcumin.

Article Snippet: The human lymphoma cell lines OCI-LY7 (KyeGEN, KGG3250-1) and SU-DHL-2 (KeyGEN, KGG3254-1) were purchased from Jiangsu KeyGEN BioTECH Co., Ltd. (Nanjing, China).

Techniques: Expressing, Concentration Assay, Western Blot, Standard Deviation, Control

Journal: Translational Cancer Research

Article Title: Curcumin inhibits the proliferation of diffuse large B-cell lymphoma by inducing ferroptosis via the ACSL4-SAT1-GPX4 axis

doi: 10.21037/tcr-2025-1-2674

Figure Lengend Snippet: Curcumin induces lipid peroxidation and loss of mitochondrial membrane potential (ΔΨm) in DLBCL cells. (A,B) Flow cytometry analysis and quantitative data of lipid reactive oxygen species levels measured by BODIPY™ C11 probe in SU-DHL-2 and OCI-LY7 cells after 24-hour treatment with the indicated concentrations of curcumin. (C,D) Flow cytometry analysis and quantitative data of mitochondrial membrane potential assessed by JC-1 staining in SU-DHL-2 and OCI-LY7 cells after curcumin treatment. The ratio of red (aggregates) to green (monomers) fluorescence is used to indicate ΔΨm. Data are presented as the mean ± standard deviation (n=3). ****, P<0.0001. DLBCL, diffuse large B-cell lymphoma.

Article Snippet: The human lymphoma cell lines OCI-LY7 (KyeGEN, KGG3250-1) and SU-DHL-2 (KeyGEN, KGG3254-1) were purchased from Jiangsu KeyGEN BioTECH Co., Ltd. (Nanjing, China).

Techniques: Membrane, Flow Cytometry, Staining, Fluorescence, Standard Deviation

Journal: Translational Cancer Research

Article Title: Curcumin inhibits the proliferation of diffuse large B-cell lymphoma by inducing ferroptosis via the ACSL4-SAT1-GPX4 axis

doi: 10.21037/tcr-2025-1-2674

Figure Lengend Snippet: Curcumin disrupts intracellular iron and redox homeostasis, promoting lipid peroxidation. (A) Intracellular Fe 2+ levels measured by the FerroOrange probe in SU-DHL-2 and OCI-LY7 cells after curcumin treatment; Hoechst 33342 indicates nuclei (200× magnification; scale bar: 50 µm). (B) Concentration of MDA, a terminal product of lipid peroxidation, in cell lysates. (C) The ratio of GSH/GSSG in cells after treatment. Data are presented as the mean ± standard deviation (n=3). *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. GSH, reduced glutathione; GSSG, oxidized glutathione; MDA, malondialdehyde.

Article Snippet: The human lymphoma cell lines OCI-LY7 (KyeGEN, KGG3250-1) and SU-DHL-2 (KeyGEN, KGG3254-1) were purchased from Jiangsu KeyGEN BioTECH Co., Ltd. (Nanjing, China).

Techniques: Concentration Assay, Standard Deviation