Journal: Cancer Cell International
Article Title: Interferon-α enhances NK cell function to counteract autologous platelet-mediated tumor immune evasion
doi: 10.1186/s12935-025-04118-w
Figure Lengend Snippet: Experimental strategy for evaluating autologous platelet effects on anti-tumor immunity of NK cells. ( A ) K562 cells were treated with platelets freshly isolated from healthy donors at a ratio of 1:500 for 3 h. Simultaneously, NK cells were purified from the same blood donors, then co-cultured with the K562 cells, which had been pre-treated with or without platelets, at a ratio of 4:1. After 16-hour incubation, we evaluated the anti-tumor responses of NK cells and the viability changes of K562 cells using flow cytometry. ( B ) Gating strategy for K562 cells. We gated on K562 cells and applied FSC-A versus FSC-H parameters to exclude doublets before analyzing CD61 signals. ( C ) CD61 + platelets on CFSE-labeled K562 cells. Representative plots (upper panel) show the percentage of CD61 + fluorescence on CFSE + K562 cells. Histograms (lower panel) display the MFI of CD61 on CFSE + K562 cells. ( D ) Summary of CD61 positivity of CFSE + K562 cells with or without platelet pre-treatment ( n = 10). ( E ) Fluorescent photographs of CD61 (PerCP-Cy5.5, red signal, excitation/emission: 488 nm/695 nm)- and CFSE (green signal, excitation/emission: 488 nm/518–521 nm) - labeled cells with or without platelets pre-treatment were taken by ImageStream ® X Mark II imaging flow cytometer. MFI, mean fluorescence intensity; **, p < 0.01
Article Snippet: Subsequently, fluorescent images of each cell were directly acquired using the ImageStream ® X Mark II imaging flow cytometer (Merck) and Confocal microscopy (Leica).
Techniques: Isolation, Purification, Cell Culture, Incubation, Flow Cytometry, Labeling, Fluorescence, Imaging
