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Fig. 5 | Journal for ImmunoTherapy of Cancer

Fig. 5

From: Targeting of CXCR3 improves anti-myeloma efficacy of adoptively transferred activated natural killer cells

Fig. 5

Long term anti-MM efficacy of activated NK cells. Activated NK cells (5-6 × 105), obtained from splenocytes of C57BL/KaLwRij, or PBS (No Cell) were i.v. transferred into MM-bearing mice 3 weeks after 5TGM1 cell injection and IL-15 was administered 18 h later. a) Average ± SEM of frequency of tumor cells in BM and spleen 7 days after adoptive transfer (n = 5 in two independent experiments). One-way ANOVA test was used to compare multiple groups. b) BM persistence of transferred activated NK cells: IL-15 or IL-12/15/18 activated CFSE+ NK cells (4 × 105) were transferred to tumor-bearing mice and donor cells were enumerated in BM and spleen 1 and 7 days later. Upper left graph shows the number of donor cells as average ± SEM of frequency of input (transferred) cell number. Student t test was performed to analyze the difference between day 1 and day 7. Upper right graph shows average MFI ± SD of CXCR4 expression levels on donor NK cells 2 and 7 days after transfer (n = 3 per group; one experiment). Lower panels, 7 days after transfer, in vivo proliferation of transferred NK cells was analyzed by CFSE dilution. Histograms were gated on NK1.1+ transferred NK cells and one representative histogram from each group is shown (n = 3 per group). Numbers in the histogram plot indicate the percentage of cells that had proliferated. c) Average MFI ± SEM of CXCR3 and CXCR4 expression levels on activated NK cells cultured in vitro for 7 days in low IL-15 concentration (10 ng/ml). d) Endogenous NK cell number was determined in BM (two tibias and femurs) by FACS analysis of CD3-NK1.1+ cells in the CFSE− population.*p < 0.05;**p < 0.01

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