Fig. 1
Dysfunction of neutrophils in TCL1 leukemia-bearing mice. a Scheme presenting the timeline of the experiment. Wild-type C57BL6/J mice were injected with splenocytes isolated from Eµ-TCL1 transgenic mouse. Next, leukemia progression was assessed by flow cytometry based on the percentage of leukemic TCL1 cells (CD5+ CD19+) among white blood cells (WBC) in the peripheral blood. The function of splenic granulocytes was analyzed at two stages of disease development: 4–10% (TCL1 early) and/or 50–80% (TCL1 late) of CD5+ CD19+ cells among WBCs in the blood. b The percentage of granulocytes (Ly6G+) among all myeloid (CD11b+) cells assessed by flow cytometry in spleens (SPL) of the control (CTR) and TCL1 leukemia-bearing mice at the indicated stages of disease development (TCL1 early, TCL1 late). The graph shows results from two independent experiments; each dot represents an individual mouse, n = 5–14, Mann‒Whitney U test, ****p ≤ 0.0001. c Representative dot plots showing the percentage of Ly6C+ Ly6G+ cells in the spleens of the control (SPL) and TCL1 leukemia-bearing mice at the indicated stages of disease development (TCL1 early, TCL1 late) assessed by flow cytometry. d The geometric mean fluorescence intensity of the oxidized CM-H2-DCFDA probe in splenic CD11b+ Ly6G+ living cells of the control (CTR) and TCL1 leukemia-bearing mice at the indicated stages of disease development (TCL1 early, TCL1 late) analyzed by flow cytometry. The graph shows results from two independent experiments; each dot represents an individual mouse, n = 5–14, Mann‒Whitney U test, **p ≤ 0.01, ***p ≤ 0.001 (left panel). Representative histograms displaying CM-H2-DCFDA fluorescence intensity versus the number of CD11b+ Ly6G+ cells in the spleens of the control (SPL) and TCL1 leukemia-bearing mice at the indicated stages of disease development (TCL1 early, TCL1 late) assessed by flow cytometry (right panel). e Graph showing the percentage of granulocytes (Ly6G+) isolated from the spleens of the control (CTR) and TCL1 leukemia-bearing mice at the early stage of the disease (TCL1) that phagocytosed Escherichia coli bioparticles. The data are presented as the mean ± SD, n = 3–4, Mann‒Whitney U test * p ≤ 0.05; f The percentage of CD63+ granulocytes (Ly6G+) isolated from the spleens of the control (CTR) and TCL1 leukemia-bearing mice at the early stage of the disease (TCL1). The graph shows the results for cells stimulated with C5a at different concentrations (10 µM and 100 µM) as a fold of that of the unstimulated control cells, n = 3–7, Mann‒Whitney U test, *p ≤ 0.05, ***p ≤ 0.001, ****p ≤ 0.0001