Journal: Stem Cell Research & Therapy

Article Title: Hypoxic TCs-preconditioned MSCs ameliorate acute lung injury via enhanced Treg recruitment and function through CXCL5/6-CXCR1 axis

doi: 10.1186/s13287-025-04858-6

Figure Lengend Snippet: CXCL5/6 knockdown attenuated the therapeutic benefits of TC supernatant-preconditioned MSCs in humanized ALI mice. A Flow cytometric analysis of CD4 + CD45 + effector T cell distribution in spleen and lung tissues across different treatment groups. N = 8. * p < 0.05, *** p < 0.001, **** p < 0.0001. ns, not significant. B Flow cytometry revealed significant alterations in pulmonary Treg proportion and their CXCR1 expression across treatment groups. N = 8. * p < 0.05, **** p < 0.0001. ns, not significant. C Comparative analysis of lung tissue homogenate cytokine profiles demonstrated differential expression of pro-inflammatory mediators ((IL-6, TNF-α, IL-17 A, IFN-γ), IL-2 and anti-inflammatory IL-10 across treatment groups. N = 8. * p < 0.05, ** p < 0.01. ns, not significant. Normal, irradiation-only mice; Control, PBMCs-only mice; ALI, PBMCs-only ALI mice; MSC, ALI mice treated with PBMCs and MSCs; TC MSC, ALI mice treated with PBMCs and TC supernatant-preconditioned MSCs; siMSC, ALI mice treated with PBMCs and CXCL5/6-knockdown MSCs preconditioned with TC supernatant

Article Snippet: Tregs were simultaneously activated and expanded by supplementing the culture with DynabeadsTM Human T-Activator CD3/CD28 (Thermo Fisher Scientific, #11456D) at a 1:1 bead-to-cell ratio and recombinant human IL-2 (1000 IU/mL; ACROBiosystems, #GMP-L02H14).

Techniques: Knockdown, Flow Cytometry, Expressing, Quantitative Proteomics, Irradiation, Control

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Development of SGN-CD47M: Protease-Activated Antibody Technology Enables Selective Tumor Targeting of the Innate Immune Checkpoint Receptor CD47

doi: 10.1158/1535-7163.MCT-24-0371

Figure Lengend Snippet: A protease-activated antibody approach using coiled-coil masking peptides improves the PK and tolerability of an antibody directed to mouse CD47. A, Masking impaired the binding of mIAP301 to mouse CD47 on HT-2 cells but could be restored upon cleavage with recombinant MMP. Symbols represent duplicate samples shown as mean ± SD. A, Decreased binding resulted in impaired RBC phagocytosis ( B ) in vitro . Representative data from three independent experiments. Symbols represent the mean ± SD from three biological replicates. *, P < 0.05; ***, P < 0.001 (one-way ANOVA followed by the Dunnett multiple comparisons test) compared with mIAP301. C, PK profiles of unmasked mIAP301 and Coil-MMP-mIAP301 in naïve BALB/c mice, which demonstrates strikingly different plasma exposure profiles because of avoidance of the large normal tissue sink for CD47. The dotted line denotes the level of quantitation in the experiment. It should be noted that the concentration of mIAP301 was only at or above the level of quantitation at the 15-minute time point. Symbols indicate n = 3 replicates and are shown as the mean ± SD. LOQ, level of quantitation. D , Antitumor activity of masked and unmasked mIAP301 antibodies was assessed in an A20 lymphoma model in BALB/c mice and showed similar antitumor activity (every two days for 8 cycles i.p. dosing). Error bars represent mean ± SD ( n = 6 mice per group). Statistical assessment for pairwise comparisons of tumor growth inhibition between treatment arms is provided in Supplementary Fig. S3. In tumor-bearing mice, masking decreased RBC depletion ( E ), and a comparison of bound antibody on RBCs vs. tumor cells provides evidence for tumor-specific mask activation ( F ). Data shown in E and F , are representative of two individual experiments. Symbols represent three biological replicates and are shown as the mean ± SD. *, P < 0.05; ***, P < 0.01; ***, P < 0.001 (one-way ANOVA followed by the Tukey multiple comparisons test) compared with mIAP301.

Article Snippet: Saturation binding ELISA experiments were performed using soluble recombinant human, mouse, or cyno CD47 (ACROBiosystems), which was diluted to an appropriate concentration in 50 mmol/L carbonate buffer at pH 9.6.

Techniques: Binding Assay, Recombinant, In Vitro, Clinical Proteomics, Quantitation Assay, Concentration Assay, Activity Assay, Inhibition, Comparison, Activation Assay

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Development of SGN-CD47M: Protease-Activated Antibody Technology Enables Selective Tumor Targeting of the Innate Immune Checkpoint Receptor CD47

doi: 10.1158/1535-7163.MCT-24-0371

Figure Lengend Snippet: SGN-CD47M is stable in human cancer whole blood and plasma. Binding of FITC-labeled hB6H12.3 and SGN-CD47M to whole blood of human patients with cancer was assessed by flow cytometry after 20 hours incubation at 37°C ( A ). SGN-CD47M stability in plasma samples of human patients with cancer was assessed using a CD47-binding ELISA ( B ) after 96 hours of incubation at 37°C. At the concentration of antibody used in the experiment (20 μg/mL), SGN-CD47M does not bind to recombinant CD47 and is not detected by ELISA. Therefore, binding indicates that the masking domain of SGN-CD47M has been proteolyzed. Under these conditions, no binding of SGN-CD47M spiked into any of the 19 samples was detected, whereas spiked control samples of hB6H12.3 yielded quantitative detection of 20 μg/mL. All samples spiked with SGN-CD47M were quantified below 350 ng/mL, which equates to <2% binding of hB6H12.3. Symbols represent two biological replicates for each donor, shown as the mean ± SD. CRC, colorectal cancer; MFI, mean fluorescent index; NSCLC, non–small cell lung cancer.

Article Snippet: Saturation binding ELISA experiments were performed using soluble recombinant human, mouse, or cyno CD47 (ACROBiosystems), which was diluted to an appropriate concentration in 50 mmol/L carbonate buffer at pH 9.6.

Techniques: Clinical Proteomics, Binding Assay, Labeling, Flow Cytometry, Incubation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Recombinant, Control

Journal: Nature Communications

Article Title: Inhibition of tau neuronal internalization using anti-tau single domain antibodies

doi: 10.1038/s41467-025-58383-4

Figure Lengend Snippet: Sensorgram (control subtracted data) of tau1N4R binding to immobilized LRP1 cluster III in the absence (black curve) or the presence of 30 (yellow curve), 100 (orange curve), and 300 nM (red curve) of ( a ) VHH E2-2, of ( b ) VHH A31, of ( c ) VHH Z70 mut1 and of ( d ) VHH H3-2.

Article Snippet: Recombinant Human LRP1 cluster III protein, Fc Tag (ACROBiosystems AG), was captured on a Serie S Protein G sensor chip (Cytiva) surface to an 1800 RU level, at a flow rate of 10 μL/min and using a working solution of 12 μg/mL in HBS-N buffer (Cytiva) supplemented with 0.005% surfactant P20 (Cytiva) and 1 mM CaCl 2 final concentration.

Techniques: Control, Binding Assay