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Purpose:
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In this experiment with isolated primary NK cells from 8 healthy human subjects.
LRS chambers from 8 healthy donors were obtained from Stanford Blood Bank. NK cells were cultured with either IL-15 (2 ng/ml), IL-2 (300 U/ml), a cocktail of IL-12 (10 ng/ml) + IL-15 (2 ng/ml) + IL-18 (50 ng/ml), or left unstimulated for 2.5 days (unstimulated, IL-2, and IL-15: n= 8; IL-12/IL-15/IL-18: n=6).
After 2.5 days, NK cells were stained with metal-labelled antibodies and signal intensities were acquired using mass cytometry (CyTOF 1)
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Conclusion:
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To analyze this data, we used several statistical and visualization tools, including viSNE (Visualization of t-Distributed Stochastic Neighbor Embedding), Citrus (Cluster identification, characterization, and regression), correspondence analysis, and the Friedman-Rafsky test.
Our analyses revealed that all three treatments (IL-2, IL-15, and IL-12/IL-15/IL-18) increase expression of CD56 and CD69. The effects of treatment with IL-2 and IL-15 are nearly indistinguishable and characterized principally by increased expression of surface markers including CD56, NKp30, NKp44, and increased expression of functional markers, such as perforin, granzyme B, and MIP-1β. The combination of IL-12/IL-15/IL-18 induces a profound shift in the repertoire structure, decreasing expression of CD16, CD57, CD8, NKp30, NKp46, and NKG2D, and dramatically increasing expression of IFN-γ.
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