target lymphoma cells  (ATCC)

Journal: bioRxiv

Article Title: Rapid receptor internalization potentiates CD7-targeted lipid nanoparticles for efficient mRNA delivery to T cells and in vivo CAR T-cell engineering

doi: 10.64898/2026.01.23.701374

Figure Lengend Snippet: (A) CAR expression in activated human T cells 24-hours after treatment with unconjugated LNP–CAR mRNA or aCD7/tLNP-CAR mRNA at doses of 0.01, 0.1, or 1 μg mRNA per million cells. (B and C) Quantification of CAR expression in CD4⁺ and CD8⁺ T cells treated with unconjugated LNP-CAR mRNA. (D and E) Quantification of CAR expression in CD4⁺ and CD8⁺ T cells treated with aCD7/tLNP-CAR mRNA. (F) Experimental scheme for evaluating the cytolytic activity of CAR T cells against target cells. (G) Representative images showing Cytotox Green fluorescence indicating Raji cell death in co-cultures with T cells treated with unmodified LNP-CAR or aCD7/tLNP-CAR at effector-to-target (E: T) ratios of 1:1, 2:1, 5:1, and 10:1. (H) Quantification of Cytotox Green fluorescence shown in (G). (I) Quantification of Raji cell viability in co-culture based on luciferase activity. n = 2 biological replicates. Data are shown as mean ± SD. Scale bar = 100 μm.

Article Snippet: To assess cytotoxic activity, CAR T cells were then co-cultured with CD20-expressing, luciferase-labeled Raji cells (ATCC, cat# CCL-86-LUC2) at effector-to-target ratios of 1:1, 2:1, 5:1, and 10:1.

Techniques: Expressing, Activity Assay, Fluorescence, Co-Culture Assay, Luciferase

Journal: bioRxiv

Article Title: Engineering a PD-L1–sensing synthetic receptor for programmable macrophage-mediated phagocytosis

doi: 10.64898/2026.01.21.700810

Figure Lengend Snippet: ( A ) Schematic representation of the CD47 “don’t eat me” signal. CD47 expressed on cancer cells engages signal regulatory protein α (SIRPα) on macrophages, thereby inhibiting phagocytosis. ( B) Flow cytometry analysis of SIRPα expression on differentiated THP-1 cells following 24 and 48 hours of polarization. Bar plots indicate the median fluorescence intensity (MFI). ( C) Flow cytometry analysis of CD47 expression across a panel of human cell lines: HEK293T (human embryonic kidney), Raji (human B-cell lymphoma), A549 (human lung carcinoma), MDA-MB-231 (human breast cancer), Jurkat (human T lymphocyte), and SKOV-3 (human ovarian adenocarcinoma). ( D) Schematic of the CD47 competition binding assay. MDA-MB-231, SKOV-3, and Jurkat cells were incubated with CV1-Fc–enriched conditioned medium prior to staining with a fluorescently labeled anti-CD47 antibody. Binding of CV1-Fc to CD47 is expected to reduce antibody binding due to competitive, steric interference. ( E-G) Representative flow cytometry histograms and bar plots showing CD47 staining of MDA-MB-231, SKOV-3, and Jurkat cells following treatment with CV1-Fc–conditioned medium (orange) or antibody-only control (red). Reduced antibody-associated fluorescence in the presence of CV1-Fc indicates effective competition for CD47 binding. n=3 ( H) Phagocytosis assay using THP-1–derived macrophages stably expressing YFP and SKOV-3 cells stably expressing mCherry. Macrophages were polarized for 48 hours and subsequently co-cultured with SKOV-3 cells for 4 hours in the presence or absence of CV1-Fc–containing conditioned medium. Double-positive YFP⁺/mCherry⁺ events represent macrophages that have engulfed cancer cells. ( I) Quantification of cancer cell engulfment by uncommitted (M0), pro-inflammatory (M1-like), and anti-inflammatory (M2-like) macrophages. Data are presented as mean ± SEM. Statistical significance was determined using paired Student’s t test. n=3 . (*p<0.05, ***p<0.001).

Article Snippet: HEK293T, MDA-MB-231, A549, THP-1, Jurkat, and Raji cell lines used in this study were purchased from the American Type Culture Collection (ATCC).

Techniques: Flow Cytometry, Expressing, Fluorescence, Binding Assay, Incubation, Staining, Labeling, Control, Phagocytosis Assay, Derivative Assay, Stable Transfection, Cell Culture

Journal: Clinical Cancer Research

Article Title: The Potential of the Gut Microbiota and Butyrate to Enhance CAR T-cell Therapy in Non-Hodgkin Lymphoma

doi: 10.1158/1078-0432.CCR-25-1676

Figure Lengend Snippet: Stimulation of CD19 CAR T cells with butyrate for 12 days increases their cytotoxic potential through phenotypic and transcriptomic changes. A, Percentage of CD69 + ( n = 3) and CD25 + ( n = 5) cells in CAR T-cell cultures stimulated with 0.5 mmol/L butyrate for 24 hours and 12 days, respectively (mean ± SEM). B, Percentage of specific lysis generated by CAR T cells and untransduced T cells (UT; mean ± SEM), stimulated with butyrate for 12 days or control, in the Raji-LucGFP cell line after 24 hours of coculture ( n = 4). C, 2ΔCt values (mean ± SEM) for IL2 , IFNG , TNF , GZMB , and PRF1 gene expression determined by qPCR in CAR T-cell cultures stimulated with butyrate for 12 days ( n = 3). D, Percentage of CAR + cells of total cells in CAR T-cell cultures stimulated with butyrate (mean ± SEM). E, Percentage of cells expressing markers of naïve (CCR7 + CD45RA + ), central memory (CM, CCR7 + CD45RA − ), effector memory (EM, CCR7 − CD45RA − ), or terminal effector memory (TEMRA, CCR7 − CD45RA + ) phenotypes in CD3 + , CD4 + , and CD8 + T-cell populations after butyrate stimulation for 12 days (mean ± SEM; n = 3). F, Percentage of T cells expressing senescence markers (CD27 − CD28 − CD57 + ; n = 3), 2ΔCt values for KLRG1 gene expression determined by qPCR ( n = 4), and fold change of mean fluorescence intensity (MFI) relative to unstained controls for the CellEvent Senescence Green probe ( n = 4; mean ± SEM) in CAR T-cell cultures stimulated with butyrate for 12 days. G, Percentage of cells (mean ± SEM) expressing the costimulatory molecule CD137 (4-1BB) in CAR T-cell cultures stimulated with butyrate for 12 days ( n = 3). H, Volcano plot and heatmap of differentially expressed genes in CAR T-cell cultures stimulated with butyrate for 12 days compared with their respective controls ( n = 4). I, Gene set enrichment analysis using Gene Ontology annotation for upregulated terms in butyrate-treated CAR T cells ( n = 4).

Article Snippet: A luminescence assay was employed to evaluate the cytotoxic potential of CAR T cells following coculture with the Raji cell line (RRID:CVCL_0511, cat. #CCL-86, ATCC), previously transduced with the ffLucGFP reporter vector.

Techniques: Lysis, Generated, Control, Gene Expression, Expressing, Fluorescence

Journal: Clinical Cancer Research

Article Title: The Potential of the Gut Microbiota and Butyrate to Enhance CAR T-cell Therapy in Non-Hodgkin Lymphoma

doi: 10.1158/1078-0432.CCR-25-1676

Figure Lengend Snippet: Oral supplementation with butyrate enhances the response to CD19 CAR T-cell treatment in vivo . A, Schematic representation of the experimental design. Monitoring and evaluation of tumor burden were performed using an in vivo imaging system (IVIS). B, In vivo bioluminescent imaging (BLI) and ( C ) survival curve of Raji-LucGFP–bearing NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice receiving CAR T cells and oral supplementation of butyrate ( n = 8) or water (control; n = 8). The remaining images are included in Supplementary Fig. S25. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: A luminescence assay was employed to evaluate the cytotoxic potential of CAR T cells following coculture with the Raji cell line (RRID:CVCL_0511, cat. #CCL-86, ATCC), previously transduced with the ffLucGFP reporter vector.

Techniques: In Vivo, In Vivo Imaging, Imaging, Control