FIGURE 5 from Characterizing Neutrophil Subtypes in Cancer Using scRNA Sequencing Demonstrates the Importance of IL1β/CXCR2 Axis in Generation of Metastasis-specific Neutrophils

• DOI: 10.1158/2767-9764.25315145.v1
• Publication Date: 2024-02-29T14:21:10Z
• Authors: Rana Fetit; Alistair S. McLaren; Mark White; Megan L. Mills; John Falconer; Xabier Cortes-Lavaud; Kathryn Gilroy; Tamsin R.M. Lannagan; Rachel A. Ridgway; Colin Nixon; Varushka Naiker; Renee Njunge; Cassie J. Clarke; Declan Whyte; Kristina Kirschner; Rene Jackstadt; Jim Norman; Leo M. Carlin; Andrew D. Campbell; Owen J. Sansom; Colin W. Steele

<p>Loss of neutrophil-specific CXCR2 attenuates suppression T-cell proliferation in neutrophils from the metastatic niche mice bearing tumors. Neutrophils liver and blood littermate either <i>Mrp8-Cre-Cxcr2<sup>+/+</sup></i> (WT) or <i>Mrp8-Cre-Cxcr2</i><sup>fl/fl</sup> (CXCR2fl) were harvested 28 days following intrasplenic injection villinCreER <i>Kras</i><sup>G12D/+</sup><i>Trp53<sup>f</sup></i><sup>l/fl</sup><i>Rosa26</i><sup>N1CD/+</sup> (KPN) cells. These placed coculture with T cells WT mice, analyzed through flow cytometry 44 hours later. <b>A,</b> Flow gating coculture. Cells gated on FSC-A/SSC-A (not shown), FCS-A/FSC-H for singlets, Live/Dead, CD3, CD4/CD8 to define CD3<sup>+</sup>, CD3<sup>+</sup>/CD4<sup>+</sup>, CD3<sup>+</sup>/CD8<sup>+</sup> populations analysis. Cell trace yellow allows tracking proliferation. The signal each division becomes subsequently dimmer, calculation number generations. Gating obtained CXCR2fl mouse (blue) unstimulated cell control (red). <b>B,</b> Proportion CD3<sup>+</sup> that proliferated tumor-bearing livers (<b>B</b>–<b>J</b>, four replicates 2 six 3 mice). <b>C,</b> Expansion index livers. <b>D,</b> Division <b>E,</b> CD4<sup>+</sup> <b>F,</b> <b>G,</b> <b>H,</b> CD8<sup>+</sup> <b>I,</b> <b>J,</b> <b>K,</b> (<b>K</b>–<b>S,</b> three <b>L,</b> mice. <b>M,</b> <b>N,</b> <b>O,</b> <b>P,</b> <b>Q,</b> <b>R,</b> <b>S,</b> *, <i>P</i> < 0.05 unpaired <i>t</i> test. **, <i>P <</i> 0.01 test.</p>