h441 lung cancer cell  (ATCC)

Journal: Cell Reports Medicine

Article Title: Lymphodepleting chemotherapy potentiates neoantigen-directed T cell therapy by enhancing antigen presentation

doi: 10.1016/j.xcrm.2025.102506

Figure Lengend Snippet: Identification of a sensitive and specific TCR targeting KRAS.G12V-A∗03:01 (A) Estimation of the prevalence of KRAS.G12V and HLA-A∗03:01 across various cancers using TCGA and CBI portal data. Data are presented as the percentage of patients estimated in this dataset. (B) HLA immunopeptidomics panel for RAS.G12V mutation, including quantification of VVVGAVGVK peptide via high-sensitivity targeted mass spectrometry. Data shown are from one biological replicate. (C) Representative flow cytometry plots of CD8 4-1BB activation (left) and IFN-γ secretion (right) from healthy donors (D98 and D104) post stimulation with mutated KRAS.G12V or WT peptides. Numbers indicate the percentage of gated events. Data are representative of two independent biological replicates, each with three technical replicates. (D) Representative flow cytometry plots showing KRAS.G12V-HLA-A∗03:01 dextramer staining of a pancreatic cancer patient that harbors the KRAS.G12V mutation and HLA-A∗03:01 (left) PBMCs and a pancreatic cancer patient without the HLA-A3 allele (right). Numbers indicate the percentage of CD8 + double-dextramer + cells. Data are representative of two independent biological replicates, each with three technical replicates. (E) Representative flow cytometry plots showing KRAS.G12V dextramer staining of D104 T cells stimulated against the WT KRAS peptide (left) or the KRAS.G12V peptide (right) in vitro . Data are representative of two independent biological replicates, each with three technical replicates. (F) Single-cell TCR sequencing clone distribution. Dextramer-double positive clones are highlighted in blue and double-negatives in gray. Data shown are from one biological replicate. (G) The table includes the sequence of the major TCR clone identified in (F). (H) Flow cytometry quantification of IFN-γ (left) and TNF-α (right)-positive TCR-T104 cells after co-culturing with KRAS.G12V or WT-pulsed B cells. Gated population from CD8 + mTCR + . Graphs represent mean ± SEM. The p value is calculated by the Mann-Whitney U test. Data are representative of five independent biological replicates, each with three technical replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (I) Flow cytometry quantification showing %4-1BB-positive TCR-T104 cells after incubation with pulsed B cells across a serial dilution of mutant or WT peptides. Data are representative of two independent biological replicates, each with three technical replicates. Each dot represents the mean of the technical replicate. The line color depicts the KRAS epitope that was pulsed (WT or mutant). (J) Luminescence mean fold change ± SEM of CD8 + Jurkat cells expressing T104-TCR across serial dilutions of WT or G12V mutant peptides pulsed on B cells. Data are representative of two independent biological replicates, each with three technical replicates. (K) Flow cytometry quantification showing %4-1BB-positive cells of TCR-T104 T cells co-cultured with endogenous lung cancer cell lines expressing the KRAS.G12V mutation (H441) or not (H3122, left), and colon cancer line (SW620) with or without A∗03:01 (right). Graphs represent mean ± SEM. The p value is calculated by the Mann-Whitney U test. Data are representative of two independent biological replicates, each with three technical replicates. (L) Flow cytometry quantification showing mean fold change ± SEM of cleaved caspase-3-mediated killing assay, using T104-TCR-T cells or irrelevant TCR (17.1.2) co-cultured with either KRAS.G12V-expressing lung cancer cell line (H441) or KRAS.Q61K cell line (Calu6). Gating on far-red-labeled+ cancer cells. The p value is calculated by the ANOVA test followed by Tukey HSD multiple correction test. Data are representative of two independent biological replicates, each with three technical replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (M) Killing dynamics of KRAS.G12V-specific T104-TCR-T cells (left), or irrelevant TCR (17.1.2 TCR, right) co-cultured with SW620-A∗03:01-GFP. GFP-positive area was normalized to time zero. Data are presented as mean ± SEM. Data are representative of four independent biological replicates, each with three technical replicates. The last time point was tested by a two-sided Student’s t test and was significant in all effector-to-target (E:T) ratios for T104 but not for irrelevant TCR ( p < 0.05). (N) Flow cytometry quantification of IFN-γ (left), TNF-α (middle), and 4-1BB (right) comparing three TCRs against 1 μg KRAS.G12V/WT-pulsed HLA-A∗03:01 B cells. Data are presented as mean ± SEM, and the p value is calculated by two-way ANOVA followed by Šidák correction for multiple comparisons. Data are representative of three independent biological replicates, each with three technical replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (O) Killing dynamics comparing three TCRs co-cultured with SW620-A∗03:01-GFP (left) and SW620-GFP (right). E:T ratio shown was maintained at 2:1 for all TCRs, normalized to the transduction rates of mTCR + cells. GFP-positive area was normalized to time zero. Data are presented as mean ± SEM. Data are representative of two independent biological replicates, each with three technical replicates. See also .

Article Snippet: The Calu6 and H441 lung cancer cell-lines were purchased from ATCC.

Techniques: Immunopeptidomics, Mutagenesis, Mass Spectrometry, Flow Cytometry, Activation Assay, Staining, In Vitro, Sequencing, Clone Assay, MANN-WHITNEY, Incubation, Serial Dilution, Expressing, Cell Culture, Labeling, Transduction

Journal: International Journal of Molecular Sciences

Article Title: Potential Application of the Myocardial Scintigraphy Agent [ 123 I]BMIPP in Colon Cancer Cell Imaging

doi: 10.3390/ijms25147747

Figure Lengend Snippet: The accumulation of [ 123 I]BMIPP and [ 18 F]FDG in ( a ) H441, ( b ) PC-14, ( c ) LS180, and ( d ) DLD-1 at 5, 10, 30, and 60 min after the respective injections ( n = 4). All cancer cell lines took up [ 123 I]BMIPP much more than [ 18 F]FDG early after its administration. The accumulation of [ 123 I]BMIPP in LS180 is the highest among the four cell lines. This result suggested that cancer cells might uptake more LCFA than glucose. † p < 0.01 and vs. [ 18 F]FDG.

Article Snippet: The human-derived lung adenocarcinoma cancer cell lines H441 (American Type Culture Collection, Manassas, VA, USA) and PC-14 (RIKEN Cell Bank, Tsukuba, Japan), and the human-derived colon cancer cell lines LS180 and DLD-1 (American Type Culture Collection, Manassas, VA, USA) were selected in this study because lung cancer and colon cancer have the number one and number two cancer mortality rates in the world, respectively [ , ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Potential Application of the Myocardial Scintigraphy Agent [ 123 I]BMIPP in Colon Cancer Cell Imaging

doi: 10.3390/ijms25147747

Figure Lengend Snippet: The accumulation of [ 123 I]BMIPP in ( a ) H441, ( b ) PC-14, ( c ) LS180, and ( d ) DLD-1 under SSO, an inhibitor of CD36, or lipofermata, an inhibitor of FATP ( n = 4). The accumulation of [ 123 I]BMIPP was significantly decreased in H441, LS180, and DLD-1 when [ 123 I]BMIPP was administered at the same time as SSO or lipofermata and taken up 5 min after. Alternatively, there was no significant difference in the accumulation of [ 123 I]BMIPP in PC-14 when the inhibitor was loaded. These results indicate that [ 123 I]BMIPP is transported into cancer cells via CD36 and FATP, with CD36 contributing more than FATP. † p < 0.01 and vs. control.

Article Snippet: The human-derived lung adenocarcinoma cancer cell lines H441 (American Type Culture Collection, Manassas, VA, USA) and PC-14 (RIKEN Cell Bank, Tsukuba, Japan), and the human-derived colon cancer cell lines LS180 and DLD-1 (American Type Culture Collection, Manassas, VA, USA) were selected in this study because lung cancer and colon cancer have the number one and number two cancer mortality rates in the world, respectively [ , ].

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Potential Application of the Myocardial Scintigraphy Agent [ 123 I]BMIPP in Colon Cancer Cell Imaging

doi: 10.3390/ijms25147747

Figure Lengend Snippet: The accumulation of [ 123 I]BMIPP in ( a ) H441, ( b ) PC-14, ( c ) LS180, and ( d ) DLD-1 at 37 °C or 4 °C at 5, 10, 30, and 60 min ( n = 4). In low-temperature conditions, the accumulation of [ 123 I]BMIPP was greatly decreased in all cell lines. Hence, [ 123 I]BMIPP also accumulates in cancer cells depending on its metabolic activity. † p < 0.01 and vs. 4 °C.

Article Snippet: The human-derived lung adenocarcinoma cancer cell lines H441 (American Type Culture Collection, Manassas, VA, USA) and PC-14 (RIKEN Cell Bank, Tsukuba, Japan), and the human-derived colon cancer cell lines LS180 and DLD-1 (American Type Culture Collection, Manassas, VA, USA) were selected in this study because lung cancer and colon cancer have the number one and number two cancer mortality rates in the world, respectively [ , ].

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Potential Application of the Myocardial Scintigraphy Agent [ 123 I]BMIPP in Colon Cancer Cell Imaging

doi: 10.3390/ijms25147747

Figure Lengend Snippet: The accumulation of [ 123 I]BMIPP in ( a ) H441, ( b ) PC-14, ( c ) LS180, and ( d ) DLD-1 under etomoxir, an inhibitor of fatty acid oxidation ( n = 4). The accumulation of [ 123 I]BMIPP significantly increased in all cell lines when [ 123 I]BMIPP was administered at the same time as etomoxir and taken up 60 min after. The accumulation of [ 123 I]BMIPP in H441 and PC-14 was greater than that in LS180 and DLD-1. Especially, the accumulation of [ 123 I]BMIPP in PC-14 after etomoxir loading was more than 8 times higher compared to the control. This result suggested that the accumulation of [ 123 I]BMIPP in cancer cells reflected FAO. † p < 0.01 and * p < 0.05 vs. control.

Article Snippet: The human-derived lung adenocarcinoma cancer cell lines H441 (American Type Culture Collection, Manassas, VA, USA) and PC-14 (RIKEN Cell Bank, Tsukuba, Japan), and the human-derived colon cancer cell lines LS180 and DLD-1 (American Type Culture Collection, Manassas, VA, USA) were selected in this study because lung cancer and colon cancer have the number one and number two cancer mortality rates in the world, respectively [ , ].

Techniques: Control