Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Expression of 5T4 and PD-L1 and anti-tumor effects of 53L10 on a panel of tumor cells. (A) The expression levels of 5T4 and PD-L1 on breast MDA-MB-231, lung A-549, gastric NCI-N87, pancreatic PANC-1, and ovarian SKOV-3 cancer cells were tested by cell ELISAs by using a commercial anti-PD-L1 or anti-5T4 antibody followed by their respective HRP-conjugated secondary Abs. The breast MCF-7 cell line was used as negative control for PD-L1 expression. Binding values were reported as the mean of determinations obtained in three independent biological experiments. Error bars depicted means ± SD (Table A). The table reports the expression levels (number of cell surface 5T4 and PD-L1) obtained by quantitative flow cytometry by QIFIKIT (Dako) analyses on MDA-MB-231, A-549, PANC-1, and MCF-7 cells. (B, C) Tumor cells were co-cultured with hPBMCs (5:1 effector-to-target cell ratio) and treated with 53L10 (B) or 53P (C) used at increasing concentrations for 48 h. The effects on tumor cell lysis were reported as dose–response curves. As a negative control, MCF-7 cells were tested. (D) Images of NCI-N87, PANC-1, and SKOV-3 cells untreated or treated with 53L10 in the presence of hPBMCs were acquired and processed by using a digital microscopy from Zen 3.2 cell discover. Cells co-cultured with hPBMCs and treated with an unrelated human IgG1 were used as a negative control. Scale bar, 200 μm.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Expressing, Negative Control, Binding Assay, Flow Cytometry, Cell Culture, Lysis, Microscopy

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Cytotoxicity and target expression levels on other breast, prostate, pancreatic, epidermoid carcinoma cell lines and CHO-transfected cells. (A) The cytotoxic activity of 53L10 was evaluated in co-cultures of various cancer cell lines and human PBMCs pre-activated with anti-CD3/anti-CD28 beads. Cells were mixed at an effector-to-target cell ratio (E/T ratio) of 5:1 or 10:1. 53L10 was added to achieve a final concentration of 10 −4 to 10 −10 M. After 3 days of culture, the amount of LDH in the culture supernatant was measured with LDH-Glo™ Cytotoxicity Assay (Promega) following the manufacturer’s instructions. Measures were performed in triplicates at each point. Cytotoxicity percentage was calculated according to the following formula: C y t o t o x i c i t y ( % ) = ( E f f e c t o r t a r g e t c e l l m i x − E f f e c t o r c e l l c o n t r o l − L o w c o n t r o l ) H i g h c o n t r o l − L o w c o n t r o l × 100 (B, C) Cytotoxic effects of 53L10 on CHO-K1 cells stably transfected with DNA encoding 5T4 or PD-L1 and expressing different levels of the targets. CHO-K1 cell lines expressing varying levels of 5T4 or PD-L1 were established and subjected to a cytotoxicity assay in the presence of 53L10 and human PBMCs pre-activated with anti-CD3/anti-CD28 beads. LDH release was evaluated after 3 days of culture. The cytotoxic activity of 53L10 against CHO-K1 cells expressing human 5T4 or human PD-L1 is shown in (B) and (C) , respectively. The number of cell surface-expressed human 5T4 or human PD-L1 was determined using the QIFIKIT (Dako) and is indicated above each graph.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Expressing, Transfection, Activity Assay, Co-Cultures, Concentration Assay, Cytotoxicity Assay, Stable Transfection

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Cell surface expression of 5T4 and PD-L1 and EC50 values for 53L10 effect on tumor cells. The values were calculated by fitting the dose–response curves reported in Figure 2 after treatment of the cells with hPBMCs from three different donors and performed in triplicates.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Analysis of tumor-infiltrating T cells in a PBMC-humanized A-549 xenograft model. (A) A-549 cells were transplanted s.c. into NOD-scid mice. When the average tumor volume reached 100 mm³, mice were treated with anti-sialo GM1 pAb. The next day, human PBMCs were introduced intraperitoneally, and 200 µg of 53P or 53L10 was subsequently administered intravenously once every 2 days. Tumors were resected on days 6 and 10. (B) Average tumor size on day 10 (mean of five mice per group). Error bars indicate standard deviations. Statistical significance was assessed by one-way ANOVA followed by Tukey’s post-hoc test and established as * p < 0.01. (C, D) On days 6 and 10, tumors from three mice per group were collected and pooled, and infiltrating lymphocytes were extracted. Human CD45 + lymphocytes were analyzed for CD4 and CD8 expression by flow cytometry.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Analysis, Xenograft Assay, Expressing, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: 53L10 T-cell engager tribody for immunotherapy of multiple solid tumors

doi: 10.3389/fimmu.2026.1934039

Figure Lengend Snippet: Comparison of the antibody constructs, tumor models, PBMC conditions, treatment schedules, endpoints, and principal findings of the previous and current studies.

Article Snippet: Human peripheral blood mononuclear cells (hPBMCs) were purchased from ATCC (PCS-800-011).

Techniques: Comparison, Construct, Tumor Models, Injection, Transplantation Assay

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: Expression of ALDH3A2 in healthy people and patients with AML (A) The expression of ALDH3A2 in various tumors compared with normal tissues. Data are represented as mean ± SEM. (B) Expression of ALDH3A2 in 88 leukemia cell lines. (C) The survival curve of patients with high and low expression of ALDH3A2 (n [low expression] = 121, n [high expression] = 30). (D) The expression of protein ALDH3A2 in HL-60, HL-60/ADM, K562, K562/ADM, and bone marrow in patients. (E) The expression of ALDH3A2 RNA in bone marrow mononuclear cells of AML patients in the CR ( n = 9), C1NR ( n = 8), and R/R ( n = 10) group. Data are represented as mean ± SEM. (F) The expression of ALDH3A2 in bone marrow mononuclear cells of 3 AML patients at their initial and recurrent stage. (G) High/low expression of ALDH3A2 and molecular mutation distribution of 66 AML patients. Data are represented as mean ± SEM. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Expressing, Mutagenesis, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: ALDH3A2-V protects AML cells from ferroptosis and cytotoxicity induced by doxorubicin (A) ALDH3A2 mRNA relative expression in HL-60/ADM and K562/ADM cells after transfection of siRNA by electroporation. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (B) The lipid peroxidation degree of HL60ADM and K562ADM cells in the control (ctrl) and ALDH3A2 knockdown (KD) groups after the same dose of doxorubicin treatment for 48 h. (C) The content of ALDH3A2 protein in HL-60, K562, U937, and KG-1α in the ALDH3A2 overexpressing (OE) group and the control (CON) group. (D) The location of ALDH3A2-V overexpression. This representative image was selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. (E) Lipid peroxidation in HL-60, K562, U937, and KG-1α in the ALDH3A2 OE group and the CON group under the same concentration of doxorubicin treatment. (F and G) Cell viability and its fitting curve of HL-60, K562, U937, and KG-1α in the ALDH3A2 OE group and the CON group under the gradient concentration of doxorubicin treatment. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Expressing, Transfection, Electroporation, Control, Knockdown, Over Expression, Concentration Assay, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: ALDH3A2 affects the sensitivity of AML cells to doxorubicin by altering 4-HNE and fatty acid content (A) The content of 4-HNE in cell lysate and culture supernatant of AML cells in the doxorubicin treatment (ADR) group and control (CON) group. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (B) The content of 4-HNE in HL-60, U937, and KG-1α of the overexpressing ALDH3A2 (OE) group and the CON group under the same concentration of doxorubicin treatment. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (C) OPLS-DA was performed on the measured fatty acids. Quantitative data for fatty acid and cell viability were collected from 3 independent experiments. (D) Variable Importance in Projection（VIP) value of different fatty acids; the difference is considered significant when the VIP>1. (E) Content of three different fatty acids in the CON and OE ALDH3A2 group. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (F) Cell viability of AML cells in heptadecanoic acid, oleic acid, and linoleic acid-pretreated groups and the CON group after the treatment of doxorubicin. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (G) Lipid peroxidation in AML cells in heptadecanoic acid-, oleic acid-, and linoleic acid-pretreated groups and the CON group after the treatment of doxorubicin. These images were selected from 3 independent replicates. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, ∗∗ indicates p < 0.01 between the two groups, and ∗∗∗ indicates p < 0.001 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Control, Concentration Assay, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: Effects of fatty acids and ALDH3A2 on plasma membrane fluidity and drug uptake (A) Laurdan staining showed different ratios of order and disorder phases in the control, heptadecanoic acid-, oleic acid-, or linoleic acid-treated groups. These representative images were selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. (B) Flow cytometry was employed to quantify Cy5 fluorescence intensity in control, heptadecanoic acid-, oleic acid-, or linoleic acid-pretreated groups at 2- and 4-h intervals following doxorubicin exposure. The representative image was selected from 3 independent replicate experiments. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (C) Laurdan staining showed different ratios of order and disorder phases in the control and ALDH3A2 high groups. The mCherry fluorescent protein was co-expressed with ALDH3A2, which indicates successful transfection of the overexpression plasmid into the cells. These representative images were selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. (D) U937 cells in the control group and ALDH3A2 high group were treated with Cy5-labeled doxorubicin. The mCherry fluorescent protein was co-expressed with ALDH3A2, which indicates successful transfection of the overexpression plasmid into the cells. These representative images were selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Clinical Proteomics, Membrane, Staining, Control, Flow Cytometry, Fluorescence, Transfection, Over Expression, Plasmid Preparation, Labeling, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: In in vivo experiments, AML with high expression of ALDH3A2 exhibits a poorer response to doxorubicin (A) Schematic diagram of the process for establishing the AML mouse model. (B) In vivo bioimaging shows the success of the model and the leukemia burden in mice before and after treatment. (C) Statistics of fluorescence values ( n = 5) on days 3 and 7 of the experiment. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (D) Survival curves of mice in different groups. (E) 4-HNE content in plasma of mice in different groups ( n = 5) on day 7. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (F) 4-HNE content in plasma of AML patients with low ( n = 5) and high ( n = 5) ALDH3A2 expression levels before and after receiving chemotherapy. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired or paired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: In Vivo, Expressing, Fluorescence, Clinical Proteomics, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: X24003 inhibits ALDH3A2 and enhances the cytotoxic effects of doxorubicin on resistant AML cells (A) OPLS-DA was performed on the measured fatty acid containment in the X24003 treatment group and control group. Quantitative data for fatty acid and cell viability were collected from 3 independent experiments. VIP value of different fatty acids; the difference is considered significant when the VIP>1. (B) X24003 exhibits a synergistic effect with doxorubicin in the elimination of doxorubicin-resistant AML cell lines HL-60ADM and K562ADM. (C) X24003 augments doxorubicin-induced lipid peroxidation in doxorubicin-resistant AML cell strains without affecting the parental cell lines. The representative image was selected from three independent replicate experiments. (D) In vivo bioimaging shows the success of the model and the leukemia burden in mice before and after treatment.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Control, In Vivo

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: HDAC2 binds to the promoter of the ALDH3A2 gene and regulates its expression (A) CUT&RUN indicates that HDAC2 binds to the promoter of the ALDH3A2 gene; the binding affinity is observed to decrease after treatment with chidamide. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (B) The expression of ALDH3A2 protein in AML cells following treatment with doxorubicin or chidamide. (C) Treatment of chidamide enhances lipid peroxidation induced by doxorubicin in AML cells. The representative image was selected from three independent replicate experiments. (D) Chidamide exhibits a synergistic effect with doxorubicin in the elimination of AML cells. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Expressing, Binding Assay, Two Tailed Test, Comparison