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The chemokine SDF-1 is a chemoattractant for human CD34+ hematopoietic progenitor cells and provides a new mechanism to explain the mobilization of CD34+ progenitors to peripheral blood.

Aiuti A, Webb IJ, Bleul C, Springer T, Gutierrez-Ramos JC - J. Exp. Med. (1997)

Bottom Line: This is the first chemoattractant reported for human CD34+ progenitor cells.Concentrations of SDF-1 that elicit chemotaxis also induce a transient elevation of cytoplasmic calcium in CD34+ cells.CD34+ cells migrating to SDF-1 include cells with a more primitive (CD34+/CD38- or CD34+/DR-) phenotype as well as CD34+ cells phenotypically committed to the erythroid, lymphoid and myeloid lineages, including functional BFU-E, CFU-GM, and CFU-MIX progenitors.

View Article: PubMed Central - PubMed

Affiliation: Center for Blood Research, Inc., Boston, Massachusetts, USA.

ABSTRACT
Hematopoietic progenitor cells migrate in vitro and in vivo towards a gradient of the chemotactic factor stromal cell-derived factor-1 (SDF-1) produced by stromal cells. This is the first chemoattractant reported for human CD34+ progenitor cells. Concentrations of SDF-1 that elicit chemotaxis also induce a transient elevation of cytoplasmic calcium in CD34+ cells. SDF-1-induced chemotaxis is inhibited by pertussis toxin, suggesting that its signaling in CD34+ cells is mediated by seven transmembrane receptors coupled to Gi proteins. CD34+ cells migrating to SDF-1 include cells with a more primitive (CD34+/CD38- or CD34+/DR-) phenotype as well as CD34+ cells phenotypically committed to the erythroid, lymphoid and myeloid lineages, including functional BFU-E, CFU-GM, and CFU-MIX progenitors. Chemotaxis of CD34+ cells in response to SDF-1 is increased by IL-3 in vitro and is lower in CD34+ progenitors from peripheral blood than in CD34+ progenitors from bone marrow, suggesting that an altered response to SDF-1 may be associated with CD34 progenitor mobilization.

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Stromal-derived SDF-1 is a chemoattractant for human and  mouse hematopoietic progenitor cells. (A) Chemotaxis assay of cord  blood (CB) CD34+ cells in response to various concentration of SDF-1  (10, 30, 100, 300, 1,000 ng/ml). Results represent the average and the  range of three experiments performed in duplicates. Data are expressed as  the percent of input cells that migrated. The percentage of migration of  CB CD34+ to undiluted MS-5 supernatant in the same experiments was  used as control and is shown as a bar diagram on the right (n = 3). (B)  Chemotactic response to SDF-1 of clonogenic progenitors (CFC). The  graph shows the number of CFU-GM, BFU-E, and CFU-MIX progenitors from human CB CD34+ cells that migrated to 300 ng/ml of SDF-1  or control media in a chemotaxis assay. (C) Transendothelial chemotaxis  in vitro of human bone marrow (BM) or mobilized peripheral blood (PB)  CD34+ cells in response to different concentration of SDF-1. Results  show the average and the range of three experiments performed in duplicates. (D) Transendothelial chemotaxis of mobilized PB CD34+ cells in  response to SDF-1 or to various chemoattractants and cytokines at the  concentrations described in the Materials and Methods. (E) Transendothelial chemotaxis in vitro of the mouse progenitor cell lines FDCP-MIX  and M1 in response to SDF-1. (F). In vivo delivery of SDF-1 into mouse  spleens increases the seeding of intravenously transplanted FDCP-MIX  cells in this organ. Experimental mice were injected intrasplenically with  SDF-1 and control mice were injected with MIP-1α or with PBS and  then transplanted i.v. with PKH-26-labeled FDCP-MIX cells, as described in Materials and Methods (three experimental mice and three  control mice per experiment; three separate experiments performed). 3 h  after the injection, mice were killed and 5 × 105 splenocytes plated in duplicate in a clonogenic assay for FDCP-MIX. After 72 h, the plates were  counted on an inverted fluorescent microscope and scored for the number of fluorescent colonies. Results show the significant increase in  FDCP-MIX clonogenic precursors per SDF-1 injected spleens relative to  control injected spleens. Data shown are from three experiments: *P <0.005.
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Figure 2: Stromal-derived SDF-1 is a chemoattractant for human and mouse hematopoietic progenitor cells. (A) Chemotaxis assay of cord blood (CB) CD34+ cells in response to various concentration of SDF-1 (10, 30, 100, 300, 1,000 ng/ml). Results represent the average and the range of three experiments performed in duplicates. Data are expressed as the percent of input cells that migrated. The percentage of migration of CB CD34+ to undiluted MS-5 supernatant in the same experiments was used as control and is shown as a bar diagram on the right (n = 3). (B) Chemotactic response to SDF-1 of clonogenic progenitors (CFC). The graph shows the number of CFU-GM, BFU-E, and CFU-MIX progenitors from human CB CD34+ cells that migrated to 300 ng/ml of SDF-1 or control media in a chemotaxis assay. (C) Transendothelial chemotaxis in vitro of human bone marrow (BM) or mobilized peripheral blood (PB) CD34+ cells in response to different concentration of SDF-1. Results show the average and the range of three experiments performed in duplicates. (D) Transendothelial chemotaxis of mobilized PB CD34+ cells in response to SDF-1 or to various chemoattractants and cytokines at the concentrations described in the Materials and Methods. (E) Transendothelial chemotaxis in vitro of the mouse progenitor cell lines FDCP-MIX and M1 in response to SDF-1. (F). In vivo delivery of SDF-1 into mouse spleens increases the seeding of intravenously transplanted FDCP-MIX cells in this organ. Experimental mice were injected intrasplenically with SDF-1 and control mice were injected with MIP-1α or with PBS and then transplanted i.v. with PKH-26-labeled FDCP-MIX cells, as described in Materials and Methods (three experimental mice and three control mice per experiment; three separate experiments performed). 3 h after the injection, mice were killed and 5 × 105 splenocytes plated in duplicate in a clonogenic assay for FDCP-MIX. After 72 h, the plates were counted on an inverted fluorescent microscope and scored for the number of fluorescent colonies. Results show the significant increase in FDCP-MIX clonogenic precursors per SDF-1 injected spleens relative to control injected spleens. Data shown are from three experiments: *P <0.005.

Mentions: In these initial experiments, it was determined that the conditioned media of MS-5 cells also contained a chemotactic activity for lymphocytes (17). Because lymphocytes are more easily obtained and isolated than CD34+ progenitor cells, they were used as indicator cells during the purification of chemoattractant activity(s) present in the conditioned media of MS-5 stromal cells. The purification of this activity was performed by sequential heparin, cation exchange, and reverse-phase HPLC chromatography. The chemoattractant activity coeluted with a single protein peak in reverse-phase HPLC and migrated as a single band in SDS-PAGE of 8 kD (17). The protein was identified by NH2-terminal sequencing (17) as SDF-1 (23–25). Electrospray and matrix-assisted laser desorption mass spectrometry gave a mass that was consistent with that predicted for SDF-1 (17). MS-5-derived SDF-1 purified to homogeneity-attracted human CD34+ cells in vitro with similar efficacy to the original MS-5-conditioned medium (Fig. 2 A). At present, we do not know whether additional factors are present in the MS-5-conditioned media that could also induce chemotaxis of CD34+ progenitor cells.


The chemokine SDF-1 is a chemoattractant for human CD34+ hematopoietic progenitor cells and provides a new mechanism to explain the mobilization of CD34+ progenitors to peripheral blood.

Aiuti A, Webb IJ, Bleul C, Springer T, Gutierrez-Ramos JC - J. Exp. Med. (1997)

Stromal-derived SDF-1 is a chemoattractant for human and  mouse hematopoietic progenitor cells. (A) Chemotaxis assay of cord  blood (CB) CD34+ cells in response to various concentration of SDF-1  (10, 30, 100, 300, 1,000 ng/ml). Results represent the average and the  range of three experiments performed in duplicates. Data are expressed as  the percent of input cells that migrated. The percentage of migration of  CB CD34+ to undiluted MS-5 supernatant in the same experiments was  used as control and is shown as a bar diagram on the right (n = 3). (B)  Chemotactic response to SDF-1 of clonogenic progenitors (CFC). The  graph shows the number of CFU-GM, BFU-E, and CFU-MIX progenitors from human CB CD34+ cells that migrated to 300 ng/ml of SDF-1  or control media in a chemotaxis assay. (C) Transendothelial chemotaxis  in vitro of human bone marrow (BM) or mobilized peripheral blood (PB)  CD34+ cells in response to different concentration of SDF-1. Results  show the average and the range of three experiments performed in duplicates. (D) Transendothelial chemotaxis of mobilized PB CD34+ cells in  response to SDF-1 or to various chemoattractants and cytokines at the  concentrations described in the Materials and Methods. (E) Transendothelial chemotaxis in vitro of the mouse progenitor cell lines FDCP-MIX  and M1 in response to SDF-1. (F). In vivo delivery of SDF-1 into mouse  spleens increases the seeding of intravenously transplanted FDCP-MIX  cells in this organ. Experimental mice were injected intrasplenically with  SDF-1 and control mice were injected with MIP-1α or with PBS and  then transplanted i.v. with PKH-26-labeled FDCP-MIX cells, as described in Materials and Methods (three experimental mice and three  control mice per experiment; three separate experiments performed). 3 h  after the injection, mice were killed and 5 × 105 splenocytes plated in duplicate in a clonogenic assay for FDCP-MIX. After 72 h, the plates were  counted on an inverted fluorescent microscope and scored for the number of fluorescent colonies. Results show the significant increase in  FDCP-MIX clonogenic precursors per SDF-1 injected spleens relative to  control injected spleens. Data shown are from three experiments: *P <0.005.
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Figure 2: Stromal-derived SDF-1 is a chemoattractant for human and mouse hematopoietic progenitor cells. (A) Chemotaxis assay of cord blood (CB) CD34+ cells in response to various concentration of SDF-1 (10, 30, 100, 300, 1,000 ng/ml). Results represent the average and the range of three experiments performed in duplicates. Data are expressed as the percent of input cells that migrated. The percentage of migration of CB CD34+ to undiluted MS-5 supernatant in the same experiments was used as control and is shown as a bar diagram on the right (n = 3). (B) Chemotactic response to SDF-1 of clonogenic progenitors (CFC). The graph shows the number of CFU-GM, BFU-E, and CFU-MIX progenitors from human CB CD34+ cells that migrated to 300 ng/ml of SDF-1 or control media in a chemotaxis assay. (C) Transendothelial chemotaxis in vitro of human bone marrow (BM) or mobilized peripheral blood (PB) CD34+ cells in response to different concentration of SDF-1. Results show the average and the range of three experiments performed in duplicates. (D) Transendothelial chemotaxis of mobilized PB CD34+ cells in response to SDF-1 or to various chemoattractants and cytokines at the concentrations described in the Materials and Methods. (E) Transendothelial chemotaxis in vitro of the mouse progenitor cell lines FDCP-MIX and M1 in response to SDF-1. (F). In vivo delivery of SDF-1 into mouse spleens increases the seeding of intravenously transplanted FDCP-MIX cells in this organ. Experimental mice were injected intrasplenically with SDF-1 and control mice were injected with MIP-1α or with PBS and then transplanted i.v. with PKH-26-labeled FDCP-MIX cells, as described in Materials and Methods (three experimental mice and three control mice per experiment; three separate experiments performed). 3 h after the injection, mice were killed and 5 × 105 splenocytes plated in duplicate in a clonogenic assay for FDCP-MIX. After 72 h, the plates were counted on an inverted fluorescent microscope and scored for the number of fluorescent colonies. Results show the significant increase in FDCP-MIX clonogenic precursors per SDF-1 injected spleens relative to control injected spleens. Data shown are from three experiments: *P <0.005.
Mentions: In these initial experiments, it was determined that the conditioned media of MS-5 cells also contained a chemotactic activity for lymphocytes (17). Because lymphocytes are more easily obtained and isolated than CD34+ progenitor cells, they were used as indicator cells during the purification of chemoattractant activity(s) present in the conditioned media of MS-5 stromal cells. The purification of this activity was performed by sequential heparin, cation exchange, and reverse-phase HPLC chromatography. The chemoattractant activity coeluted with a single protein peak in reverse-phase HPLC and migrated as a single band in SDS-PAGE of 8 kD (17). The protein was identified by NH2-terminal sequencing (17) as SDF-1 (23–25). Electrospray and matrix-assisted laser desorption mass spectrometry gave a mass that was consistent with that predicted for SDF-1 (17). MS-5-derived SDF-1 purified to homogeneity-attracted human CD34+ cells in vitro with similar efficacy to the original MS-5-conditioned medium (Fig. 2 A). At present, we do not know whether additional factors are present in the MS-5-conditioned media that could also induce chemotaxis of CD34+ progenitor cells.

Bottom Line: This is the first chemoattractant reported for human CD34+ progenitor cells.Concentrations of SDF-1 that elicit chemotaxis also induce a transient elevation of cytoplasmic calcium in CD34+ cells.CD34+ cells migrating to SDF-1 include cells with a more primitive (CD34+/CD38- or CD34+/DR-) phenotype as well as CD34+ cells phenotypically committed to the erythroid, lymphoid and myeloid lineages, including functional BFU-E, CFU-GM, and CFU-MIX progenitors.

View Article: PubMed Central - PubMed

Affiliation: Center for Blood Research, Inc., Boston, Massachusetts, USA.

ABSTRACT
Hematopoietic progenitor cells migrate in vitro and in vivo towards a gradient of the chemotactic factor stromal cell-derived factor-1 (SDF-1) produced by stromal cells. This is the first chemoattractant reported for human CD34+ progenitor cells. Concentrations of SDF-1 that elicit chemotaxis also induce a transient elevation of cytoplasmic calcium in CD34+ cells. SDF-1-induced chemotaxis is inhibited by pertussis toxin, suggesting that its signaling in CD34+ cells is mediated by seven transmembrane receptors coupled to Gi proteins. CD34+ cells migrating to SDF-1 include cells with a more primitive (CD34+/CD38- or CD34+/DR-) phenotype as well as CD34+ cells phenotypically committed to the erythroid, lymphoid and myeloid lineages, including functional BFU-E, CFU-GM, and CFU-MIX progenitors. Chemotaxis of CD34+ cells in response to SDF-1 is increased by IL-3 in vitro and is lower in CD34+ progenitors from peripheral blood than in CD34+ progenitors from bone marrow, suggesting that an altered response to SDF-1 may be associated with CD34 progenitor mobilization.

Show MeSH
Related in: MedlinePlus