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

Journal: Journal of Translational Medicine

Article Title: Exercise-mobilized lymphocytes enhance antibody-based immunotherapy in multiple myeloma through CD16 + NK cell–mediated cytotoxicity

doi: 10.1186/s12967-026-07888-7

Figure Lengend Snippet: Schematic detailing the experimental design. The workflow illustrates the collection of lymphocytes from participants, followed by ex vivo co-culture with myeloma cells under pre-treatment and co-treatment conditions with therapeutic regimens. ( A ) Overview of the clinical exercise protocol. Participants completed a screening visit followed by the main exercise trial. Peripheral blood was collected at rest, during, and 1-hour post-exercise to isolate and cryopreserve PBMCs for subsequent analysis. ( B ) Ex vivo treatment and co-culture workflow. Effector cells (PBMCs or isolated NK cells) and target cells (MM1.S and MM1.R) were incubated overnight under specific conditions. Following incubation and washout, effectors and targets were co-cultured to assess cytotoxicity (with or without CD16 blockade), immune phenotype, and tumor ligand expression. Abbreviations: DRd: Daratumumab, lenalidomide, and Dexamethasone; MRd: Magrolimab, lenalidomide, and Dexamethasone

Article Snippet: Briefly, 100 μL of EDTA whole blood was incubated with the following antibodies CD8-VioBlue, CD57-VioBlue, CD14-VioGreen, CD3-VioGreen, CD62L-FITC, NKG2C-FITC, TCR-Vd2-FITC, CD4-PE, NKG2D-PE, CD45-PE-Vio615, CD20-Pe-Vio770, CD45RA-PerCPVio770, NKG2A-Pe-Vio770, CD16-APC, KLRG1-APC, and CD56-APC-Vio770 (Miltenyi Biotec Inc., Gernany) for 30 min at room temperature.

Techniques: Ex Vivo, Co-Culture Assay, Isolation, Incubation, Cell Culture, Expressing

Journal: Journal of Translational Medicine

Article Title: Exercise-mobilized lymphocytes enhance antibody-based immunotherapy in multiple myeloma through CD16 + NK cell–mediated cytotoxicity

doi: 10.1186/s12967-026-07888-7

Figure Lengend Snippet: Exercise-mobilized NK cells display enhanced cytotoxic phenotypes and differential responses to DRd vs MRd. ( A ) Baseline CD38% positive and MFI) and CD47% positive) on NK cells measured in peripheral blood mononuclear cells (PBMCs) collected at rest (REST; red) or during acute exercise at 80% VO2max (EX; blue). ( B ) NK-cell frequency (% of CD45+ lymphocytes) after overnight IL-15 culture under control, DRd (daratumumab + lenalidomide + dexamethasone), or MRd (magrolimab + lenalidomide + dexamethasone) conditions. Representative CD3 vs. CD56 dot plots are shown on the right for REST and EX across regimens, with the NK-cell gate (CD3−CD56+) indicated. ( C ) Expression of monoclonal-antibody–associated markers after overnight culture: CD38% positive and MFI) under control vs. MRd conditions, and CD47% positive and MFI) under control vs. DRd conditions. ( D ) Fc-gamma receptor (FcγR) profiles after overnight culture, showing the expression of CD16 and CD32% positive and MFI). ( E ) Expression of activating and cytotoxic markers after overnight culture: NKG2C, CD57, NKG2D, and DNAM-1% positive and MFI). In all bar graphs, red bars represent REST and blue bars represent EX. Bars show the mean ± SEM with paired donor values overlaid and connected by dotted lines. PBMCs from N = 10 healthy donors were used. Each condition was run in duplicate, and the results were averaged per donor. NK cells were defined as CD3−CD56+ lymphocytes. Statistical tests were two-sided. Panel A: paired t-test (EX vs. REST). Panel B: one-way repeated-measures ANOVA within each exercise state (REST or EX), followed by Dunnett’s multiple comparisons test vs. The control group. Panels C–E: two-way repeated-measures ANOVA with factors for exercise (REST vs. EX) and regimen (e.g., control vs. DRd), followed by Tukey’s multiple comparisons test.Significance is indicated by asterisks or exact p -values where appropriate. Thresholds were set at * p < 0.05, ** p < 0.01, and *** p < 0.001. “ns” denotes not significant. Abbreviations: PBMC, peripheral blood mononuclear cell; NK, natural killer; MFI, mean fluorescence intensity; DRd, daratumumab/lenalidomide/dexamethasone; MRd, magrolimab/lenalidomide/dexamethasone

Article Snippet: Briefly, 100 μL of EDTA whole blood was incubated with the following antibodies CD8-VioBlue, CD57-VioBlue, CD14-VioGreen, CD3-VioGreen, CD62L-FITC, NKG2C-FITC, TCR-Vd2-FITC, CD4-PE, NKG2D-PE, CD45-PE-Vio615, CD20-Pe-Vio770, CD45RA-PerCPVio770, NKG2A-Pe-Vio770, CD16-APC, KLRG1-APC, and CD56-APC-Vio770 (Miltenyi Biotec Inc., Gernany) for 30 min at room temperature.

Techniques: Control, Expressing, Fluorescence

Journal: Journal of Translational Medicine

Article Title: Exercise-mobilized lymphocytes enhance antibody-based immunotherapy in multiple myeloma through CD16 + NK cell–mediated cytotoxicity

doi: 10.1186/s12967-026-07888-7

Figure Lengend Snippet: Exercise enhances purified NK cell cytotoxicity via a CD16-dependent mechanism. ( A ) Pre-treatment: target cells were exposed to the drug regimens before co-culture with purified natural killer (NK) cells. ( B ) Co-treatment: both target cells and purified NK cells were exposed to the drug regimen overnight before co-culture. ( C ) Pre-treatment: target cells were exposed to antibodies before being co-cultured with NK cells. ( D ) Co-treatment: both target cells and NK cells were exposed to antibodies overnight prior to co-culture. ( A - B ) The bar graphs show the percentage of specific lysis for control conditions, combination therapies DRd (daratumumab + lenalidomide + dexamethasone) and MRd (magrolimab + lenalidomide + dexamethasone), and their respective monoclonal antibodies alone, daratumumab ( D ) and magrolimab (M). ( C - D ) Assays were run with control (no antibody), daratumumab alone ( D ), and magrolimab alone (M). The effect of CD16 blockade was tested in the “D (anti-CD16)” and “M (anti-CD16)” groups.Red bars represent NK cells collected at rest (rest), while blue bars represent NK cells collected during acute exercise (exercise) at 80% VO2max. Data are presented as mean ± SEM. Individual dots represent data from each donor ( N = 6), with lines connecting paired rest and exercise samples. Statistical comparisons shown are for the effect of exercise (exercise vs. Rest) within each treatment group. Significance is indicated as * p < 0.05; “ns” denotes not significant. The analysis was performed using a two-way repeated-measures ANOVA for pre- and co-treatment conditions separately (factors: exercise and regimen), followed by Tukey’s multiple comparisons test. Abbreviations: D, daratumumab; DRd, daratumumab/lenalidomide/dexamethasone; M, magrolimab; MRd, magrolimab/lenalidomide/dexamethasone

Article Snippet: Briefly, 100 μL of EDTA whole blood was incubated with the following antibodies CD8-VioBlue, CD57-VioBlue, CD14-VioGreen, CD3-VioGreen, CD62L-FITC, NKG2C-FITC, TCR-Vd2-FITC, CD4-PE, NKG2D-PE, CD45-PE-Vio615, CD20-Pe-Vio770, CD45RA-PerCPVio770, NKG2A-Pe-Vio770, CD16-APC, KLRG1-APC, and CD56-APC-Vio770 (Miltenyi Biotec Inc., Gernany) for 30 min at room temperature.

Techniques: Purification, Co-Culture Assay, Cell Culture, Lysis, Control, Bioprocessing

Journal: Frontiers in Immunology

Article Title: Osteoclasts affect the anti-cancer activity of NK cells

doi: 10.3389/fimmu.2026.1730283

Figure Lengend Snippet: Characterization of NK cells in presence of OCs and A549s by flow cytometry. Box and whiskers plots show the % of CD56 + CD16 + (A) and CD56 ++ CD16 - NK cells (B) , the expression on NK cells of NK activator receptors DNAM-1 (C) , NKp44 (D) , NKp30 (E) , activation marker CD69 (F) and of inhibitory receptors TIM3 (G) and TIGIT (H) in co-culture with A549s, OCs and both OCs+A549s. Groups were compared using a one-way repeated measures analysis of variance with post hoc Tukey test (* p < 0.05; ** p < 0.01).

Article Snippet: We also used commercially available mAbs as CD69 PE, CD3 PercP, CD16 APC-Vio770, TIGIT (Miltenyi Biotec, Bergisch Gladbach, Germany), CD56 PE-Cy7 (anti-human Beckaman Coulter, Fullerton, CA, USA), and LEAF anti-TIM3 (345004, Biolegend, San Diego, California).

Techniques: Flow Cytometry, Expressing, Activation Assay, Marker, Co-Culture Assay