Figure 2.
Figure 2. Dominant role of CXCR4/SDF-1 and Gi-signals in BM homing of ex vivo cytokine-incubated BM-HPCs. (A) SCF incubation increases cell motility. Spontaneous migration (▦) and SDF-1-directed chemotactic migration (▪) of fresh or overnight SCF-incubated cells, with or without PTX, was quantified after 4 hours. Depicted are mean plus SEM. Spontaneous and SDF-1-directed migration were both significantly increased by SCF incubation. This increase in migration was completely blocked by PTX, to levels of spontaneous migration of fresh BM, which apparently represents Gi-protein-independent migration (*P < .05). (B-D) BM homing of overnight SCF-incubated BM-HPCs. BM homing of SCF-incubated BM-HPCs was tested 3 hours (B) or 18 hours (C,D) after transplantation of lethally irradiated recipients. Donor-cell source/treatment and recipient strain are indicated below the respective bars. BM homing is given as percentage of injected CFU-Cs recovered per femur. Depicted are mean plus SEM of all mice tested with this donor/host constellation. (B) Three-hour BM homing of SCF-incubated cells treated with or without AMD3100. BM homing of fresh WT BM cells was significantly reduced by AMD3100 incubation/coinjection (*P < .05). (C) Eighteen-hour BM homing of SCF-incubated BM cells. BM homing of α4-/- cells in WT recipients was significantly reduced compared with WT-to-WT (*P < .05). BM homing of WT cells in CD62-/- recipients or of β2-/- cells in WT recipients were no different from WT-to-WT. (D) Eighteen-hour BM homing of SCF plus PTX-treated BM cells. BM homing of SCF plus PTX-treated WT cells in WT recipients was 75% reduced compared with SCF-treated WT-to-WT without PTX (P < .05). BM homing of SCF plus PTX-treated α4-/- cells in WT recipients or BM homing of SCF plus PTX-treated WT cells in VCAM-1-/- hosts was almost completely abrogated, significantly more strongly than SCF plus PTX-treated WT-to-WT (*P < .05). The inhibition of homing of SCF plus PTX-incubated β2-/- cells in WT recipients was similar to that of SCF plus PTX-treated WT donor cells.

Dominant role of CXCR4/SDF-1 and Gi-signals in BM homing of ex vivo cytokine-incubated BM-HPCs. (A) SCF incubation increases cell motility. Spontaneous migration (▦) and SDF-1-directed chemotactic migration (▪) of fresh or overnight SCF-incubated cells, with or without PTX, was quantified after 4 hours. Depicted are mean plus SEM. Spontaneous and SDF-1-directed migration were both significantly increased by SCF incubation. This increase in migration was completely blocked by PTX, to levels of spontaneous migration of fresh BM, which apparently represents Gi-protein-independent migration (*P < .05). (B-D) BM homing of overnight SCF-incubated BM-HPCs. BM homing of SCF-incubated BM-HPCs was tested 3 hours (B) or 18 hours (C,D) after transplantation of lethally irradiated recipients. Donor-cell source/treatment and recipient strain are indicated below the respective bars. BM homing is given as percentage of injected CFU-Cs recovered per femur. Depicted are mean plus SEM of all mice tested with this donor/host constellation. (B) Three-hour BM homing of SCF-incubated cells treated with or without AMD3100. BM homing of fresh WT BM cells was significantly reduced by AMD3100 incubation/coinjection (*P < .05). (C) Eighteen-hour BM homing of SCF-incubated BM cells. BM homing of α4-/- cells in WT recipients was significantly reduced compared with WT-to-WT (*P < .05). BM homing of WT cells in CD62-/- recipients or of β2-/- cells in WT recipients were no different from WT-to-WT. (D) Eighteen-hour BM homing of SCF plus PTX-treated BM cells. BM homing of SCF plus PTX-treated WT cells in WT recipients was 75% reduced compared with SCF-treated WT-to-WT without PTX (P < .05). BM homing of SCF plus PTX-treated α4-/- cells in WT recipients or BM homing of SCF plus PTX-treated WT cells in VCAM-1-/- hosts was almost completely abrogated, significantly more strongly than SCF plus PTX-treated WT-to-WT (*P < .05). The inhibition of homing of SCF plus PTX-incubated β2-/- cells in WT recipients was similar to that of SCF plus PTX-treated WT donor cells.

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