Fig. 4: Tumor tissues with elevated scIgGs had a reduced NK cell infiltration and decreased antitumor cytotoxic activities.
From: Impairment of IgG Fc functions promotes tumor progression and suppresses NK cell antitumor actions
a Tumor infiltrations of NK cells (CD45+/CD3-/CD49b+) in scIgG containing tumors (4T1-IdeS) were significantly lower than that in the counterpart control tumors (4T1), n = 5, p < 0.05. Percentage of NK cells in tumors was measured by flow cytometry analysis after tumors were dissociated into single cells as described in the Method. b Infiltration of NK cells in scIgG tumors (BT474-IdeS) was lower than that in control BT474 tumors. NK cells (CD49b+) by IHC staining was analyzed using image J software. Three tumor slides from individual mouse tumors were stained and 10 images per tumor slides were taken for each group. Mean ± SD of relative staining intensities (RI) in each tumor groups are compared, n = 30, **p < 0.005. Levels of granzyme B in scIgG containing tumors were significantly reduced than that in their counterpart control tumors, 4T1-IdeS vs 4T1 in (c) and BT474-IdeS vs BT474 in (d). IHC staining method was used and tumor staining intensities were quantified using image J software, n = 20, p < 0.05. Perforin levels in tumors containing scIgGs were lower than that in the control group using IF staining method. A representative image for each group is shown in (e) for BT474-IdeS vs BT474 and (f) for 4T1-IdeS vs 4T1 tumor group. The IF stained intensities in tumors were quantified using image J software. The graphs show the mean of immunofluorescence intensity and error bars indicate SD, n = 20, **p < 0.005. The scIgG containing 4T1-IdeS tumors had significantly lower percentage of NK cells with perforin (PFN + ) expression (g) and granzyme B (GrB + ) expression (h) when compared with that in control tumors. Percentages of NK cells with perforin or granzyme B positive staining were analyzed using the Boolean combination gating method with FlowJo software, n = 5, p < 0.05. i NKp46 expression levels scIgG containing tumors were reduced when compared with that in the counterpart controls. A representative IF image is shown for 4T1-IdeS and 4T1 control group, n = 5 for each mouse group. j The IF staining intensities of NKp46 were quantified as fluorescence pixel units (FPU) and 40 images (n = 40, from 5 tumor slides) using image j software, **p < 0.01.
