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