aml cell lines thp1  (DSMZ)

Journal: Nature Communications

Article Title: Allogeneic CD33-directed CAR-NKT cells for the treatment of bone marrow-resident myeloid malignancies

doi: 10.1038/s41467-025-56270-6

Figure Lengend Snippet: a – e Studying the in vitro antitumor efficacy of Allo15 CAR33-NKT cells against human AML cell lines. CAR33-T cells and non-CAR33-engineered PBMC-T cells were included as therapeutic cell controls. a Experimental design. b Schematics showing the indicated human AML cell lines. THP1-FG, THP1 cell line engineered to overexpress the firefly luciferase and green fluorescence protein dual reporters (FG); KG1-FG, KG1 cell line engineered to overexpress FG; HL60-FG, HL60 cell line engineered to overexpress FG; THP1-FG CD33-/- , THP1-FG cell line further engineered to knockout the CD33 gene; THP1-FG CD1d-/- , THP1-FG cell line further engineered to knockout the CD1d gene; THP1-FG CD33/CD1d-/- , THP1-FG cell line further engineered to knockout the CD33 and CD1d genes. c FACS detection of CD33 and CD1d expressions on the indicated AML cells. d Heatmap showing the NKR ligand expressions on the indicated AML cells. The number represents the percentage of NKR ligand-positive tumor cells out of the total tumor cells. Three independent tumor cell samples were analyzed, and the average numbers are presented. e Tumor cell killing data at 24 h ( n = 4 from four different cell product donors). f , g Studying the tumor cell killing mechanisms of Allo15 CAR33-NKT cells mediated by NKRs (i.e., NKG2D and DNAM-1). f Experimental design. g Tumor cell killing data at 24 h (E:T ratio = 10:1; n = 4 from four different cell product donors). h Diagram showing the CAR/TCR/NKR triple tumor-targeting mechanisms of Allo15 CAR33-NKT cells, and the CAR single tumor-targeting mechanism of CAR33-T cells. GrzB, Granzyme B. Created in BioRender. FANG, Y. (2025) https://BioRender.com/j50y057 . i , j Studying the expression of effector molecules of Allo15 CAR33-NKT cells. i FACS detection of surface CD69 as well as intracellular Perforin and Granzyme B in Allo15 CAR33-NKT cells. j Quantification of ( i ) ( n = 3 from three different cell product donors). Representative of 3 experiments. Data are presented as the mean ± SEM. ns not significant, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, by one-way ANOVA ( g , j ), or two-way ANOVA ( e ). Source data and exact p values are provided as a file.

Article Snippet: Human AML cell line THP1 (cat. no. TIB-202), KG1 (cat. no. CCL-246), and HL60 (cat. no. CCL-240) were purchased from the American Type Culture Collection (ATCC).

Techniques: In Vitro, Luciferase, Fluorescence, Knock-Out, Expressing

Journal: Nature Communications

Article Title: Allogeneic CD33-directed CAR-NKT cells for the treatment of bone marrow-resident myeloid malignancies

doi: 10.1038/s41467-025-56270-6

Figure Lengend Snippet: a – g Studying the in vivo antitumor efficacy of Allo15 CAR33-NKT cells in a THP1-FG human AML xenograft NSG mouse model. a Experimental design. b BLI images. c Quantification of ( b ) ( n = 5). d Kaplan–Meier survival curves ( n = 5). e BLI images showing the presence of residual tumor cells in mouse tissues at the termination day. GI tract, gastrointestinal tract. FACS analyses of surface CD33 ( f ) and intranuclear cancer stem cell (CSC) marker ( g ) expression in THP1-FG tumor cells, collected from mouse bone marrow at the termination day ( n = 5). h – n Studying the in vivo antitumor efficacy of Allo15 CAR33-NKT cells in a KG1-FG human AML xenograft NSG mouse model. h Experimental design. i BLI images. j Quantification of ( i ) ( n = 5). k Kaplan–Meier survival curves ( n = 5). l BLI images showing the presence of residual tumor cells in mouse tissues at the termination day. FACS analyses of surface CD33 ( m ) and intranuclear CSC marker ( n ) expression in KG1-FG tumor cells collected from mouse bone marrow at the termination day ( n = 5). o – r Studying the antitumor efficacy of Allo15 CAR33-NKT cells against primary patient samples. o Experimental design. Created in BioRender. LI, Y. (2025) https://BioRender.com/o37h997 p Blast cell killing data at 24 h ( n = 4 from four different therapeutic cell donors). Data from three AML and one MDS patient samples are presented. FACS analyses of surface CD33 ( q ) and intranuclear CSC marker r expression in the remaining blast cells collected after the 24-h in vitro tumor cell killing assay ( n = 4 from four different therapeutic cell donors). s Diagram illustrating the ability of Allo15 CAR33-NKT cells to target CD33-low/negative LSPCs in myeloid malignancies. Created in BioRender. LI, Y. (2025) https://BioRender.com/m06k581 . Representative of 2 ( a – n ) and 3 ( o – r ) experiments. Data are presented as the mean ± SEM. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, by two-tailed Student’s t test ( f , g , m , n ), one-way ANOVA ( c , j , p , q ), or log rank (Mantel-Cox) text adjusted for multiple comparisons ( d , k ). Source data and exact p values are provided as a file.

Article Snippet: Human AML cell line THP1 (cat. no. TIB-202), KG1 (cat. no. CCL-246), and HL60 (cat. no. CCL-240) were purchased from the American Type Culture Collection (ATCC).

Techniques: In Vivo, Marker, Expressing, In Vitro, Two Tailed Test

Journal: Nature Communications

Article Title: Allogeneic CD33-directed CAR-NKT cells for the treatment of bone marrow-resident myeloid malignancies

doi: 10.1038/s41467-025-56270-6

Figure Lengend Snippet: a – f Studying the synergistic effect of Allo15 CAR33-NKT cells with HMA Decitabine using an in vitro tumor cell killing assay. a Experiment design. THP1-D, THP1 tumor cells treated with decitabine. b FACS detection of CAR target (CD33), iNKT TCR target (CD1d), and NKR targets (i.e., CD112, MICA/B, ULBP-1, and ULBP-2/5/6) on the indicated AML cells. c Quantification of ( b ) ( n = 4 from four different experimental batches). d Tumor cell killing data at 24 h ( n = 4 from four different experimental batches). e FACS detection of intracellular Granzyme B production by Allo15 CAR33-NKT cells. f Quantification of ( e ) ( n = 4 from four different experimental batches). g Diagram showing the upregulation of CD1d and NK ligands on AML tumor cells following treatment with HMA. Created in BioRender. FANG, Y. (2025) https://BioRender.com/n85n160 . h – k Studying the in vivo synergistic effect of Allo15 CAR33-NKT cells with HMA Decitabine using a THP1-FG human AML xenograft NSG mouse model. h Experimental design. i BLI images showing the presence of tumor cells in experimental mice over time. j Quantification of ( i ) ( n = 5). k Kaplan–Meier survival curves of experimental mice over time ( n = 5). Representative of 2 ( h – k ) and 3 ( a – g ) experiments. Data are presented as the mean ± SEM. ns not significant, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, by two-tailed Student’s t test ( c ), one-way ANOVA ( d , f , j ), or log rank (Mantel-Cox) text adjusted for multiple comparisons ( k ). Source data and exact p values are provided as a a file.

Article Snippet: Human AML cell line THP1 (cat. no. TIB-202), KG1 (cat. no. CCL-246), and HL60 (cat. no. CCL-240) were purchased from the American Type Culture Collection (ATCC).

Techniques: In Vitro, In Vivo, Two Tailed Test

Journal: Nature Communications

Article Title: Allogeneic CD33-directed CAR-NKT cells for the treatment of bone marrow-resident myeloid malignancies

doi: 10.1038/s41467-025-56270-6

Figure Lengend Snippet: a , b Studying the graft-versus-host (GvH) response of Allo15 CAR33-NKT cells using an in vitro mixed lymphocyte reaction (MLR) assay. CD33-negative PBMCs were pre-sorted using MACS or FACS and used as stimulator cells. Conventional CAR33-T cells were included as responder controls. a Experimental design. b ELISA analyses of IFN-γ production on day 4. N, no addition of stimulator cells ( n = 4). c – h Studying the GvHD risk of Allo15 CAR33-NKT cells using a human xenograft NSG mouse model. c Experimental design. d Clinical GvHD score recorded over time ( n = 5). The score was calculated as the sum of individual scores of 6 categories (body weight, activity, posture, skin thickening, diarrhea, and dishevelment; score 0–2 for each category). p was calculated using Day 50 data. e Body weight measured over time ( n = 5). p was calculated using Day 50 data. f Kaplan–Meier survival curves ( n = 5). g H&E-stained tissue sections. Tissues were collected from experimental mice on day 50. Scale bar, 100 µm. h Quantification of ( g ) ( n = 5). i – l Studying the CRS response induced by Allo15 CAR33-NKT cells using a THP1-FG human AML xenograft NSG mouse model. i Experimental design. j Body weight of experimental mice over time ( n = 4). ELISA analyses of mouse IL-6 and SAA3 in mouse serum ( k ) or peritoneal fluid ( l ) ( n = 4). NT, samples collected from tumor-bearing mice receiving no therapeutic cell treatment. m Studying the long-term safety of Allo15 CAR33-NKT cells using a human xenograft NSG mouse model. Tissues from experimental mice were collected 120 days after injection with Allo15 CAR33-NKT cells. Data were presented as pathologist’s scores of individual mouse tissues ( n = 5). Representative of 1 ( k ) and 3 ( a – j ) experiments. Data are presented as the mean ± SEM. ns not significant, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, by Student’s t test ( d , e , h ), one-way ANOVA ( b , k , l ), or log rank (Mantel-Cox) text adjusted for multiple comparisons ( f ). Source data and exact p values are provided as a file.

Article Snippet: Human AML cell line THP1 (cat. no. TIB-202), KG1 (cat. no. CCL-246), and HL60 (cat. no. CCL-240) were purchased from the American Type Culture Collection (ATCC).

Techniques: In Vitro, Mlr Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Staining, Injection

Journal: Nature Communications

Article Title: Allogeneic CD33-directed CAR-NKT cells for the treatment of bone marrow-resident myeloid malignancies

doi: 10.1038/s41467-025-56270-6

Figure Lengend Snippet: a – d Generation of 15 CAR33-T cells. a Diagram showing the design of Lenti/CAR33-IL-15 lentivector, and the generation of 15 CAR33-T cells from healthy donor PBMCs. Created in BioRender. LI, Y. (2025) https://BioRender.com/o37h997 b FACS detection of the CAR33 and CD4/CD8 co-receptors on CAR33-T and 15 CAR33-T cells. c ELISA analyses of IL-15 production by CAR33-T and 15 CAR33-T cells cultured in vitro for 24 h ( n = 3; n indicates different PBMC donors). Note the successful incorporation and expression pf IL-15 transgene in the 15 CAR33-T cells. d Yield of CAR33-T and 15 CAR33-T cells ( n = 3; n indicates different PBMC donors). e , f Studying the antitumor efficacy of 15 CAR33-T cells against human AML cell lines. e Experimental design. f Tumor cell killing data at 24 h ( n = 4). g – j Studying the in vivo antitumor efficacy of 15 CAR33-T cells using a THP1-FG human AML xenograft NSG mouse model. g Experimental design. h BLI images showing the presence of tumor cells in experimental mice over time. i Quantification of ( h ) ( n = 5). j Kaplan–Meier survival curves of experimental mice over time ( n = 5). – n Studying the in vivo antitumor efficacy of 15 CAR33-T cells using a KG1-FG human AML xenograft NSG mouse model. Experimental design. l BLI images showing the presence of tumor cells in experimental mice over time. m Quantification of ( l ) ( n = 5). n Kaplan–Meier survival curves of experimental mice over time ( n = 5). Note that the data for the Vehicle, Allo15 CAR33-NKT, and CAR33-T groups were also presented in the main Figs. h– . Representative of 2 ( g – n ) and 3 ( a – f ) experiments. Data are presented as the mean ± SEM. ns, not significant; **** p < 0.0001, by Student’s t test ( c , d ), one-way ANOVA ( i , m ), or two-way ANOVA ( f ). Source data and exact p values are provided as a file.

Article Snippet: Human AML cell line THP1 (cat. no. TIB-202), KG1 (cat. no. CCL-246), and HL60 (cat. no. CCL-240) were purchased from the American Type Culture Collection (ATCC).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, In Vitro, Expressing, In Vivo

Journal: Scientific Reports

Article Title: CD157 signaling promotes survival of acute myeloid leukemia cells and modulates sensitivity to cytarabine through regulation of anti-apoptotic Mcl-1

doi: 10.1038/s41598-021-00733-5

Figure Lengend Snippet: CD157 modulates cellular stress responses in AML cell lines. ( A ) OCI-AML3 cells were cultured overnight in the absence of FCS, then the SY11B5 anti-CD157 mAb or mIgG (both at 10 µg/ml) were added for 24 h. Cell viability was measured by AnnexinV/PI staining and flow cytometry analysis. Histograms show the effect of nutrient deprivation on cell viability expressed as fold change of viable cells compared to cells maintained in standard culture conditions, and are the mean ± SEM of six independent experiments performed in quadruplicate. * p < 0.05, *** p < 0.001, ns = not significant, one-way ANOVA with Tukey’s multiple comparison test. ( B ) OCI-AML3 were cultured overnight in serum-free medium then, SY11B5 or RF3 mAbs to CD157 (10 µg/ml) were added for 24 h. Whole cell lysates were subjected to Western blotting and probed with the indicated antibodies. Blots were re-probed using antibodies against the total proteins. β-Actin was used as loading control. Numbers below blots indicate fold change in the expression of each protein relative to untreated control, normalized to the corresponding β-actin and total protein (mTOR and AKT). Uncropped images are provided in Supplementary Fig. S9 ( C ) CD157-high or low OCI-AML3, THP1 and U937 cells were cultured in standard conditions or in serum-free medium for 24 h, and then subjected to AnnexinV/PI staining and flow cytometry analysis. Results are expressed as fold change of viable cells normalized to CD157-high cells maintained in standard culture conditions and are the mean ± SEM of three experiments. * p < 0.05, ** p < 0.01, one-way ANOVA with Tukey’s multiple comparison test. ( D ) CD157-high and CD157-low THP1 cells and ( E ) CD157-high and CD157-low U937 cells were maintained for 18 h in FCS-free culture medium, then were stimulated with FCS for 10 min or 2 h. Whole cell lysates were subjected to Western blotting and probed with the indicated antibodies. Blots were re-probed using antibodies against the total proteins. β-Actin was used as loading control. Numbers below blots indicate fold change in the expression of each protein relative to untreated CD157-high cells, normalized to the corresponding β-actin and total protein (AKT and GSK3β). Panels ( D ) and ( E ) show one Western blot representative of three independent experiments performed. Uncropped blot images are provided in Supplementary Fig. S9.

Article Snippet: U937 and THP1 AML cell lines (from American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in RPMI-1640 medium supplemented with 10% FCS; OCI-AML3 cells (kindly provided by G. Saglio, University of Torino, Italy) were cultured in DMEM medium supplemented with 20% FCS.

Techniques: Cell Culture, Staining, Flow Cytometry, Comparison, Western Blot, Control, Expressing

Journal: Scientific Reports

Article Title: CD157 signaling promotes survival of acute myeloid leukemia cells and modulates sensitivity to cytarabine through regulation of anti-apoptotic Mcl-1

doi: 10.1038/s41598-021-00733-5

Figure Lengend Snippet: CD157 knockdown increases AraC-induced cell death in U937 cells. ( A ) THP1 (black line), OCI-AML3 (dashed black line) or U937 (red line) cells were treated with increasing concentrations of AraC for 24 h to assess the sensitivity of each AML cell line to AraC. ( B ) The sensitivity of CD157-high (red line) versus CD157-low (blue line) U937 cells to AraC was compared by Presto Blue assays. Six replicates for each condition have been performed. Results are the mean ± SEM of four experiments. **** p < 0.0001, Student’s t-test. The EC 50 for U937 cells with high or low CD157 was calculated using the GraphPad software (see “ ”). ( C ) Representative dot plot (left) and histogram (right) of CD157-high and CD157-low U937 cells treated with AraC for 24 h, obtained by flow cytometry analysis after AnnexinV/PI staining. Histograms show the mean ± SEM of eight independent experiments. ** p < 0.01, two-way ANOVA with Sidak’s multiple comparison test. ( D ) U937/CD157-high and U937/CD157-low were treated with AraC for 24 h, then whole cell lysate was subjected to Western blotting. Numbers below blots indicate fold change in the expression of each protein relative to untreated CD157-high cells, normalized to the corresponding β-actin and full length protein (PARP, Bax and Caspase-3). β-Actin was used as loading control. Left panels: the levels of Mcl-1, pGSK-3β, cBax, cPARP, and cCaspase-3 were determined by densitometry analysis and are the mean of three experiments run in parallel. Uncropped blot is provided in Supplementary Fig. S10. Data are expressed as the mean ± SEM of band densities of western blots from three separate experiments. * p < 0.05, ** p < 0.01, **** p < 0.0001, ns = not significant, two-way ANOVA with Sidak’s multiple comparison test.

Article Snippet: U937 and THP1 AML cell lines (from American Type Culture Collection, ATCC, Manassas, VA, USA) were grown in RPMI-1640 medium supplemented with 10% FCS; OCI-AML3 cells (kindly provided by G. Saglio, University of Torino, Italy) were cultured in DMEM medium supplemented with 20% FCS.

Techniques: Knockdown, Software, Flow Cytometry, Staining, Comparison, Western Blot, Expressing, Control

Journal: Frontiers in Immunology

Article Title: Decitabine-Mediated Epigenetic Reprograming Enhances Anti-leukemia Efficacy of CD123-Targeted Chimeric Antigen Receptor T-Cells

doi: 10.3389/fimmu.2020.01787

Figure Lengend Snippet: DAC enhances anti-leukemia activity of CD123 CAR-T cells in vitro and in vivo . (A) CD123 CAR-T cells, generated from four patients and three healthy donors, pretreated with different concentrations of DAC for 48 h, and then were co-cultured with THP1 cells at different E: T ratio (Effector: Target) for 4 h after drug wash-out. LDH release assay was used to detect the cytotoxicity of CD123 CAR-T cells. Three independent experiments were conducted. Mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. (B) CD123 CAR-T cells were treated with different doses of DAC for 48 h and apoptosis was determined by flow cytometry. (C) NSG mice bearing AML tumor xenografts constructed by THP1-luciferase cells were randomized to receive one of the following treatments on 5 consecutive days: PBS, CD123 CAR-T cells (2.5 × 10 6 ), DAC (1mg/kg), or CD123 CAR-T cells (2.5 × 10 6 ) combined with DAC (1mg/kg). (D) Tumor signals were monitored with Lumina imaging on day 9, day12, and day 19. (E) The bioluminescence signal was measured as total photon flux normalized for exposure time and surface area and expressed in units of photons (p) per second per cm 2 per steradian (sr) ( n = 4 mice in each group).

Article Snippet: The human acute myeloid leukemia (AML) cell line THP1 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured as described previously ( ).

Techniques: Activity Assay, In Vitro, In Vivo, Generated, Cell Culture, Lactate Dehydrogenase Assay, Flow Cytometry, Construct, Luciferase, Imaging

Journal: Frontiers in Immunology

Article Title: Decitabine-Mediated Epigenetic Reprograming Enhances Anti-leukemia Efficacy of CD123-Targeted Chimeric Antigen Receptor T-Cells

doi: 10.3389/fimmu.2020.01787

Figure Lengend Snippet: Investigations on the effect of DAC on intrinsic potency and subsets of CAR-T cells. CD123 CAR-T cells generated from two patients and three healthy donors were treated with or without 1 μM DAC for 48 h. Then RNA seq gene expression analysis was performed. (A) The top 200 genes in either direction were shown. (B) Heat map of T cell pathways enriched in genes upregulated or downregulated in DAC pretreated CD123 CAR-T cells. For each pathway, a single sample enrichment score was calculated by the expression level of FRKM of each gene (genes of each T cell subsets detailed in ), and the mean was taken by group. Each column represented an individual sample pretreated with or without DAC, and each row represented the enrichment of gene expression profiles involved in different subsets of T cells. A color gradient represents the changes of mean normalized enrichment score from red (highest) to blue (lowest) (ranging from −2 to +2). Blue indicates a down-regulation and Red indicates an up-regulation. (C) Enrichment of T cell pathways in CD123 CAR-T cells were presented. Each point represents a specimen. Red for patients and green for healthy donors. Mean ± SD. (D) CD123 CAR-T cells ( n = 3) were treated with a series of doses of DAC for 48 h with the presence of THP1 at E:T = 1:1. T cell subsets (Tcm: CD45RA–CCR7+; Tnaive: CD45RA+CCR7+; Tem: CD45RA–CCR7–; T EMRA : CD45RA+CCR7–) were measured by FACS analysis. Three independent experiments were conducted. Mean ± SD. * p < 0.05, ** p < 0.01.

Article Snippet: The human acute myeloid leukemia (AML) cell line THP1 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured as described previously ( ).

Techniques: Generated, RNA Sequencing, Gene Expression, Expressing

Journal: Cell

Article Title: Inhibition of Dihydroorotate Dehydrogenase Overcomes Differentiation Blockade in Acute Myeloid Leukemia

doi: 10.1016/j.cell.2016.08.057

Figure Lengend Snippet: (A) THP1 cells were implanted subcutaneously in the flank of SCID mice; treatment with brequinar (BRQ) decreased tumor growth. QD, once every day.

Article Snippet: Cell Culture The U937 and THP1 human AML cell lines, as well as all other human AML cell lines, were purchased from the American Type Culture Collection (ATCC; http://www.atcc.org ).

Techniques:

Journal: Cell

Article Title: Inhibition of Dihydroorotate Dehydrogenase Overcomes Differentiation Blockade in Acute Myeloid Leukemia

doi: 10.1016/j.cell.2016.08.057

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cell Culture The U937 and THP1 human AML cell lines, as well as all other human AML cell lines, were purchased from the American Type Culture Collection (ATCC; http://www.atcc.org ).

Techniques: Recombinant, Expressing, Knock-In, Software