1. CD44-Mediated Adhesiveness of Human Hematopoietic Progenitors to Hyaluronan Is Modulated by Cytokines
by Stéphane Legras, Jean-Pierre Lévesque, Rachida Charrad, Kohji Morimoto, Caroline Le Bousse, Denis Clay, Claude Jasmin, and Florence Smadja-Joffe
Blood
Volume 89(6):
March 15, 1997
©1997 by American Society of Hematology

2. Opposite effects of anti-CD44 MoAbs J173 and H90 on spontaneous adhesion of CD34+ cells to immobilized HA. 51Cr-labeled CD34+ cells (2 × 105 cells/mL) were incubated at 4°C for 15 minutes with 5 μg/mL of IgG1, J173, or H90 MoAbs in the cell adhesion medium ...
Opposite effects of anti-CD44 MoAbs J173 and H90 on spontaneous adhesion of CD34+ cells to immobilized HA. 51Cr-labeled CD34+ cells (2 × 105 cells/mL) were incubated at 4°C for 15 minutes with 5 μg/mL of IgG1, J173, or H90 MoAbs in the cell adhesion medium and put in contact for 15 minutes at 37°C with immobilized HA in 96-well culture plate (100 μL per well, 3 wells per point). After discarding the nonadherent cells by two washes, the adherent cell-associated radioactivity was measured using a β scintillator counter as described in the Materials and Methods. Data, expressed as the percentage of the radioactivity of the input, are the means ± 1 SE calculated from five independent experiments.
Stéphane Legras et al. Blood 1997;89:
©1997 by American Society of Hematology

3. Cell surface expression of CD44 on input, HA-adherent and HA-nonadherent CD34+ cells.
Cell surface expression of CD44 on input, HA-adherent and HA-nonadherent CD34+ cells. CD34+ cells were stained with FITC-conjugated anti-CD44 MoAb J173. Negative controls (grey lines) are cells labeled with FITC-conjugated IgG1. Flow cytometric analysis was performed as described in the Materials and Methods. Data are from a representative experiment. Four independent experiments gave similar results.
Stéphane Legras et al. Blood 1997;89:
©1997 by American Society of Hematology

4. Surface expression of CD44 on CD34+ cells treated with PMA, H90 MoAb, or cytokines.
Surface expression of CD44 on CD34+ cells treated with PMA, H90 MoAb, or cytokines. Cells were treated with either 10−7 mol/L PMA for 16 hours, H90 MoAb (5 μg/mL), or cytokines SCF (10 ng/mL), GM-CSF (0.1 ng/mL), or IL-3 (10 ng/mL) for 15 minutes at 37°C as described in the Materials and Methods. Control cells were either untreated cells or cells treated with IgG1. Thereafter, cells were washed and labeled with FITC-conjugated J173 MoAb. Negative controls (dotted lines) were treated cells labeled with FITC-conjugated IgG1. Flow cytometry analysis was performed as described in the Materials and Methods.
Stéphane Legras et al. Blood 1997;89:
©1997 by American Society of Hematology

5. Cell surface expression of CD38 and HLA-DR II on input and HA-adherent CD34+ HPC. CD34+ HPC were stained with MoAbs to CD38 (PE-conjugated) and to HLA-DR (FITC-conjugated).
Cell surface expression of CD38 and HLA-DR II on input and HA-adherent CD34+ HPC. CD34+ HPC were stained with MoAbs to CD38 (PE-conjugated) and to HLA-DR (FITC-conjugated). Negative controls (grey lines) are cells labeled with PE-conjugated IgG1 or FITC-conjugated IgG2a. Flow cytometric analysis was performed as described in the Materials and Methods. Data are from a representative experiment. Four independent experiments gave similar results.
Stéphane Legras et al. Blood 1997;89:
©1997 by American Society of Hematology

6. Dose-dependency of cytokine-stimulating effect on CD34+ HPC adhesion to HA. Adhesion assays were performed in the presence of 0.1 to 10 ng/mL of (•) SCF, (▪) IL-3, or (▴) GM-CSF.
Dose-dependency of cytokine-stimulating effect on CD34+ HPC adhesion to HA. Adhesion assays were performed in the presence of 0.1 to 10 ng/mL of (•) SCF, (▪) IL-3, or (▴) GM-CSF. Cell adhesion was measured at 15 minutes as described in the Materials and Methods. Data are means ± 1 SE calculated from three independent experiments.
Stéphane Legras et al. Blood 1997;89:
©1997 by American Society of Hematology

7. Time-dependency of cytokine-stimulating effect on CD34+ cell adhesion to HA. Adhesion assays were performed in the presence of 10 ng/mL (•) SCF and (▪) IL-3 and 0.1 ng/mL of (▴) GM-CSF.
Time-dependency of cytokine-stimulating effect on CD34+ cell adhesion to HA. Adhesion assays were performed in the presence of 10 ng/mL (•) SCF and (▪) IL-3 and 0.1 ng/mL of (▴) GM-CSF. (♦) Cells preincubated at 4°C for 15 minutes with 5 μg/mL of the anti-CD44 MoAb H90. (▾) Cells treated with the same dose of IgG1 in the same conditions. Cell adhesion was measured at 15 minutes, 30 minutes, and 2 hours. Measures were performed in triplicate as indicated in the legend to Fig 1. Data are means ± 1 SE calculated from three independent experiments.
Stéphane Legras et al. Blood 1997;89:
©1997 by American Society of Hematology