Figure 5.

The presence of tumor does not influence the phenotype, potential for degranulation, and cytokine production of adoptively transferred NK cells. (A) A scheme is shown for evaluation of expanded NK cells in vivo. Mice were irradiated and received human SK-MEL-28 melanoma cells (with tumor) or PBS (w/o tumor) by intravenous (i.v.) injection. Three days later after establishment of tumor, all mice were treated (i.v.) with human NK cells expanded with the optimized protocol. Three days after transfer of the NK cells, cells from the blood and lungs of the mice were recovered. (B) NK cells from the blood and lungs were enumerated and NK cell numbers per g of lung and per mL of blood are shown. (C) NK cells from mouse lungs as described in A were analyzed for the surface markers NKG2D, TRAIL, and DNAM-1 by flow cytometry and shown are the mean fluorescence intensity (MFI) corrected by isotype subtraction. (D) NK cells as described in A were stimulated with PMA/Iono and analyzed by flow cytometry for degranulation and production of IFNγ and TNF-α. Means and standard deviations for NK cells from three independent donors are shown with cells from 3–5 mice that were pooled per condition for each donor. Statistical significance was tested by paired Student's t-test.