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. 2022 Nov 8;13:6752. doi: 10.1038/s41467-022-34467-3

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© The Author(s) 2022, corrected publication 2022

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Fig. 5 — A Pearson correlation between CD15^high neutrophils and GZMK^high CD8⁺ T_EM (n = 22; p = 0.0387) B Representative contour plots of GZMK expression in CD8⁺ T_EM in LN and HN patients. C Frequency of GZMK^high CD8⁺ T_EM cells in LN (n = 7) and HN (n = 14, p = 0.009) patients. D GZMK Mean Fluorescence Intensity (MFI) in HN (n = 9) and LN (n = 8) tumors. Scale bar 10 μm. Pearson correlation between: CXCR2⁺ neutrophils and GZMK^high CD8⁺ T cells (E, n = 22, p < 0.0001); CXCR2⁺ neutrophils and tumor size (F, n = 22, p = 0.0002), GZMK^high CD8⁺ T cells and tumor size (G, n = 21, p = 0.0004) within tumor (T) of MC38 tumor-bearing mice. GZMK^high CD8⁺ T frequency within T (H, p = 0.050) and tumor weight (I, p = 0.030) in mice upon αLy6G antibody-mediated neutrophil’s depletion (n = 6) or treatment with isotype control (n = 5). J Representative confocal images and quantification of GZMK MFI in CD8⁺ and CD8⁻ (not-CD8) cells within a 10 μm distance from neutrophils (CD66b⁺). Scale bar 5 μm. K Representative confocal images and normalized quantification (see “Methods”) of GZMK MFI in CD8⁺ within 10 μm (Touching, n = 161) or between 10 and 20 μm (Near, n = 203) from neutrophils (CD66b⁺). Yellow dotted lines represent 20 μm (left) and 10 μm (right). Scale bar 10 μm. L. GZMK⁺ MFI within activated CD8⁺ T cells (CTR) co-cultured at 1:1 ratio with neutrophils isolated from Bone Marrow (BM, n = 9) or MC38 tumors (T, n = 3) (p = 0.015 for CTR vs 1:1 NEU T, p < 0.0001 for CTR vs 1:1 NEU BM. M. Frequency of GZMK^high CD8⁺ T cells (CTR) after co-culture with HD neutrophils (n = 16), either in plate or in Transwell (TRANS). The CD8⁺ T cells-neutrophils ratios are indicated (p = 0.011 for CTR vs 1:1, p = 0.098 for CTR vs 1:10, p = 0.976 for CTR vs 1:1 TRANS). N. Co-culture of CD8⁺ isolated from patients’ PB with neutrophils isolated from tumor (NEU T) (n = 2), CD15^high and CD15^lowneutrophils. Bars represent mean ± SEM or box and whisker plots indicate Min to Max value, two-tailed paired t test (C, L, M, N), two-tailed Mann–Whitney test (H), two-tailed unpaired t test (I). Source data are provided as a Source Data file.