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