Journal: iScience

Article Title: From byproduct to biotherapeutic: Comparative study of buffy coats and leukoreduction system chambers for NK cell-based immunotherapies

doi: 10.1016/j.isci.2026.114907

Figure Lengend Snippet: Functional characterization of NK cells (A) Degranulation capacity of NK cells following a 2 h co-culture with K562 leukemia cells, measured by surface CD107a expression ( n = 5). (B and C) Europium-based cytotoxicity assay assessing the specific lysis of K562 target cells at E:T ratios of 10:1, 5:1, 3:1, 1:1, and 0.5:1 on day 7 (B) and day 14 (C) of culture to monitor functional decline over time ( n = 6 for both time points). Here, all comparisons were not significant and are summarized as such (n.s.: p > 0.05). (D) IFN-γ release following 16 h of co-culture of NK cells with K562 cells, quantified via Luminex assay ( n = 5). (E) Kinetic live-cell killing assay using the Incucyte monitoring of Nuclight Red signal loss in K562 cells over 72 h at an E:T ratio of 1:1 ( n = 6). (F) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis thresholds during Incucyte co-culture with K562 cells. (G) CD107a degranulation assay of NK cells with or without pre-incubation with trastuzumab (anti-HER2 antibody) during a 2 h co-culture with MDA-MB-453 cells ( n = 5). (H) Incucyte-based cytotoxicity assay shows the real-time lysis of MDA-MB-453 cells by trastuzumab-loaded NK cells at an E:T ratio of 1:1 over 72 h ( n = 6). (I) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis during the ADCC assay. (n varies as not all samples reached the benchmark values within the observation period). All ADCC experiments have been performed after 14 days of in vitro NK cell expansion. N/A indicates that statistical analysis was not possible for this group. 9J) CD107a degranulation of unmodified NK cells and ErbB2-CAR-NK cells after 2 h co-culture with MDA-MB-453 cells ( n = 5). (K) Incucyte-based cytotoxicity assay showing killing dynamics of ErbB2-CAR-NK cells against MDA-MB-453 cells at an E:T ratio of 1:1 over 72 h ( n = 6). (L) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis during the CAR-mediated cytotoxicity assay. (n varies as not all samples reached the benchmark values within the observation period). All CAR-killing experiments have been performed after 14 days of in vitro NK cell expansion. Unless otherwise stated, n refers to biologically independent donor samples and is indicated per group. Data are shown as mean ± SEM. p-values were determined by Student’s t test with Welsh correction (A, D, F, G, I, J, and L) or by one-way ANOVA (B and C) with Tukey post-test (K and L).

Article Snippet: Expression of the target antigen ErbB2 on cells used in cytotoxicity assays was determined by flow cytometry using an ErbB2 APC antibody (Clone REA522; Miltenyi, Bergisch Gladbach, Germany).

Techniques: Functional Assay, Co-Culture Assay, Expressing, Cytotoxicity Assay, Lysis, Luminex, Degranulation Assay, Incubation, ADCC Assay, In Vitro

Journal: Cell Reports Medicine

Article Title: Valrubicin-loaded immunoliposomes targeting antigens on immunosuppressive cells to circumvent resistance to cancer immunotherapy

doi: 10.1016/j.xcrm.2026.102632

Figure Lengend Snippet: Val-ILs and αPD-1 affect the growth and spread of breast tumor cells 10 6 4T1 cells were transplanted through i.d. injection into the mammary gland of female BALB/c mice. When tumors reached 50 mm 3 , mice were i.v. injected with Val-ILs (10 12 NPs) and i.p. injected with αPD-1 (200 μg) on days 6, 9, and 12. Analyses were performed on day 15 (A–E). (A) Tumor growth volumes measured in vivo. (B) Image of tumors isolated ex vivo from the mammary gland of mice (two independent in vivo experiments). Val-ILs-Combo was efficient in reducing the tumor volumes, and the combination of Val-ILs-Combo and αPD-1 was even more efficient. Mice without breast tumors are identified with a black cross. (C) FC on TAM cells in the TME and macrophages in the spleen, to identify M1-like and M2-like phenotypes. (D) Images of mice measured with in vivo imaging system technology showing the detection of bioluminescent 4T1 cells in the mammary gland, following treatment with Val-ILs-Combo and/or αPD-1. (E) FC was used to detect metastatic CD44 + GFP + 4T1 cells spreading to distant organs following treatment. Examples of FC plots and a heatmap displaying the mean percentages. Undetected (ud) indicates the absence of detectable tumor 4T1 cells. (F) On day 30, tumor-free mice were rechallenged with a secondary injection of 10 6 4T1 cells into the contralateral mammary gland. Tumor growth was monitored to evaluate the establishment of a protective anti-tumor memory. The number of mice used per group is indicated on the figure, data are shown as means ± SD, and p values are compared to Val-ILs-IgG + αIgG or between the different groups and are calculated using one-way ANOVA with Tukey’s multiple comparisons test. See also .

Article Snippet: APC anti-Mouse CD44 (Clone REA664) , Miltenyi Biotec , Cat# 130-119-121; RRID: AB_2751628.

Techniques: Injection, In Vivo, Isolation, Ex Vivo, In Vivo Imaging