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. 2009 Mar 25;4(3):e4942. doi: 10.1371/journal.pone.0004942

Figure 3. Human Dex harbour functional IL-15Rα and synergize with IL-15 for NK cell proliferation in vitro and IFNγ production in vitro.

Figure 3

A. Immunoblotting of IL-15Rα from Dex and DC lysates. Western Blot analysis on 10–40 µg of protein lysates obtained from immature DC (iDC), Dex or Tex (exosomes from Mel888 melanoma cell line) using anti-IL-15Rα mAb. Positive controls included anti-HLA-DRα, -TSG 101 and -HSC 70 Abs. Representative immunoblots of two normal volunteers are depicted (NV1 and NV2). Molecular weights are indicated on the left lane. B. Proliferative effects of recombinant IL-15 and Dex on NK cells. CFSE-labeled NK cells were cultured with or without 10 µg autologous Dex or allogenic Tex in complete medium containing 0.5 ng/ml of human recombinant IL-15. At day 6 of culture, NK cell proliferation was determined by flow cytometry and the number of divisions were counted and depicted. A representative experiment out of two is shown. C–D. Synergistic effects between Dex and recombinant IL-15 for NK cell triggering. NV's PBL were cultured without (white histograms) or with (black histograms) 10 µg autologous Dex and increasing concentrations of human recombinant IL-15. NK (CD56+ CD3−) cells were then analysed for CD69 expression by flow cytometry (C) or supernatants were harvested to measure IFNγ levels in EIA (D). The graphs depict means±SEM of % of CD69 expressing NK cells in 3 experiments (C) or IFNγ concentrations in 4 experiments (D). * p<0.05, ** p<0.01 and ns: non significant.