Figure 2.
Short-term and long-term granulocyte depletion reduces inflammation and prolongs survival in the ITK-SYK PTCL mouse model. (A) Workflow for short-term granulocyte depletion with the Ly6G antibody vs control IgG antibody. Antibody injection (250 μg) was performed IP on day 14 and day 28 after ITK-SYK transplantation. (B) Percentage of granulocytes and monocytes in the PB 1 day after control or Ly6G antibody treatment (n = 6 per group). (C) Percentage of different T-cell subsets analyzed by FACS within ITK-SYK/GFP+ and GFP− thymus cells of ITK-SYK mice on day 28 after first and day 14 after second antibody injection (n = 6 per group). (D) Phenotypic score, spleen weight, and liver weight of ITK-SYK mice treated with Ly6G vs control antibody, day 14 after the second treatment (n = 6 per group). (E) Representative images of ITK-SYK mice treated with IgG control vs Ly6G antibody. (F) Kaplan-Meier survival curve representing the survival of ITK-SYK mice after 2 IP injections with IgG isotype control antibody or anti-Ly-6G antibody (n = 6 per group). (G) Genotype of long-term neutrophil granulocyte depleted Lysm-Cre/WT/MCL-1fl/fl mice (NP) on a C57BL/6N background. (H) Total granulocytes and monocytes from the PB of WT and NP mice that had received transplantation with ITK-SYK, day 55 after transplantation (n = 6 per group). (I) Percentage of neutrophil granulocytes (CD11b+ Ly6G+) in the BM and spleen of WT control or NP mice. (J) Percentage of different T-cell subsets analyzed by FACS within ITK-SYK/GFP+ and GFP– thymus cells of ITK-SYK mice with WT vs Lysm-Cre/WT/MCL-1fl/fl background (n = 6 per group). (K) Phenotypic score of WT ITK-SYK mice vs (NP) ITK-SYK mice on day 35 after transplantation (n = 6 per group). (L) Kaplan-Meier survival curve representing the survival of WT ITK-SYK mice vs Lysm-Cre/WT/MCL-1fl/fl (NP) ITK-SYK mice (n = 6 per group). (B-D,H-J) Bars represent mean values with error bars showing the SEM. Statistical significance was calculated using Student unpaired t test. Asterisks represent the significance of the difference between GFP+ and GFP– populations within the same mice. (F,L) Statistical analysis was performed using the log-rank Mantel-Cox test. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. IgG, immunoglobulin G; SEM, standard error of the mean.

Short-term and long-term granulocyte depletion reduces inflammation and prolongs survival in the ITK-SYK PTCL mouse model. (A) Workflow for short-term granulocyte depletion with the Ly6G antibody vs control IgG antibody. Antibody injection (250 μg) was performed IP on day 14 and day 28 after ITK-SYK transplantation. (B) Percentage of granulocytes and monocytes in the PB 1 day after control or Ly6G antibody treatment (n = 6 per group). (C) Percentage of different T-cell subsets analyzed by FACS within ITK-SYK/GFP+ and GFP− thymus cells of ITK-SYK mice on day 28 after first and day 14 after second antibody injection (n = 6 per group). (D) Phenotypic score, spleen weight, and liver weight of ITK-SYK mice treated with Ly6G vs control antibody, day 14 after the second treatment (n = 6 per group). (E) Representative images of ITK-SYK mice treated with IgG control vs Ly6G antibody. (F) Kaplan-Meier survival curve representing the survival of ITK-SYK mice after 2 IP injections with IgG isotype control antibody or anti-Ly-6G antibody (n = 6 per group). (G) Genotype of long-term neutrophil granulocyte depleted Lysm-Cre/WT/MCL-1fl/fl mice (NP) on a C57BL/6N background. (H) Total granulocytes and monocytes from the PB of WT and NP mice that had received transplantation with ITK-SYK, day 55 after transplantation (n = 6 per group). (I) Percentage of neutrophil granulocytes (CD11b+ Ly6G+) in the BM and spleen of WT control or NP mice. (J) Percentage of different T-cell subsets analyzed by FACS within ITK-SYK/GFP+ and GFP– thymus cells of ITK-SYK mice with WT vs Lysm-Cre/WT/MCL-1fl/fl background (n = 6 per group). (K) Phenotypic score of WT ITK-SYK mice vs (NP) ITK-SYK mice on day 35 after transplantation (n = 6 per group). (L) Kaplan-Meier survival curve representing the survival of WT ITK-SYK mice vs Lysm-Cre/WT/MCL-1fl/fl (NP) ITK-SYK mice (n = 6 per group). (B-D,H-J) Bars represent mean values with error bars showing the SEM. Statistical significance was calculated using Student unpaired t test. Asterisks represent the significance of the difference between GFP+ and GFP– populations within the same mice. (F,L) Statistical analysis was performed using the log-rank Mantel-Cox test. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. IgG, immunoglobulin G; SEM, standard error of the mean.

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