Potential Role for Hyaluronan and the Hyaluronan Receptor RHAMM in Mobilization and Trafficking of Hematopoietic Progenitor Cells by Linda M. Pilarski,

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Potential Role for Hyaluronan and the Hyaluronan Receptor RHAMM in Mobilization and Trafficking of Hematopoietic Progenitor Cells by Linda M. Pilarski, Eva Pruski, Juanita Wizniak, Darlene Paine, Karen Seeberger, Michael J. Mant, Christopher B. Brown, and Andrew R. Belch Blood Volume 93(9):2918-2927 May 1, 1999 ©1999 by American Society of Hematology

Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

For HPCs from mobilized blood, intracellular HA receptors correlate with HA binding, but surface HA receptors do not.r 2 is the regression coefficient when HA binding is set as the dependent variable. r sis the Spearman rank order correlation coefficient th... For HPCs from mobilized blood, intracellular HA receptors correlate with HA binding, but surface HA receptors do not.r 2 is the regression coefficient when HA binding is set as the dependent variable. r sis the Spearman rank order correlation coefficient that does not assume normal distribution and does not require assigning a dependent variable. Data points indicate individual patient samples assayed for both variables. Aliquots of the same samples were analyzed for all parameters in this figure. Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

HA binding by permeabilized mobilized blood HPCs is inhibited by anti-RHAMM and by anti-CD44. HA binding by permeabilized mobilized blood HPCs is inhibited by anti-RHAMM and by anti-CD44. Mobilized blood HPCs were treated with MoAb before HA binding as in Fig 1. The relative decrease was calculated as the MFI of HA binding after pretreatment with anti-RHAMM or anti-CD44 divided by the MFI after anti-β1 integrin. For all 3 samples, the same pattern of MoAb inhibition was observed. Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

Ratios of ic:sRHAMM and ic:sHA binding for mobilized BM HPCs correlate with those of paired mobilized blood HPCs collected at the same point in time. Ratios of ic:sRHAMM and ic:sHA binding for mobilized BM HPCs correlate with those of paired mobilized blood HPCs collected at the same point in time. For each of 4 mobilized BL/BM pairs, the ratio of icRHAMM MFI:sRHAMM MFI or of icHA binding MFI:sHA binding MFI was analyzed by linear regression with sRHAMM or sHA binding as the dependent variable. (•) Ratios for RHAMM; (○) ratios for HA binding. Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

In steady-state BM HPCs, but not in mobilized blood HPCs, the MFI of sRHAMM is strongly correlated with the MFI of icRHAMM. In steady-state BM HPCs, but not in mobilized blood HPCs, the MFI of sRHAMM is strongly correlated with the MFI of icRHAMM. The MFI of sRHAMM and icRHAMM for mobilized blood HPCs (top panel) or steady-state BM HPCs (bottom panel) were analyzed by linear regression analysis and Spearman rank order correlation. The bottom panel shows the 95% confidence limits. Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

HA binding by mobilized blood HPCs is increased when cell metabolism, lipid rafts, or cytoskeletal assembly are inhibited. HA binding by mobilized blood HPCs is increased when cell metabolism, lipid rafts, or cytoskeletal assembly are inhibited. Mobilized blood HPCs from 6 different individuals were treated with sodium azide, nystatin, or cytochalasin B as indicated in Materials and Methods, followed by the addition of HA-FITC and then MoAbs to stain HPCs. The same pattern was observed for all 6 samples. The percentage of increase above basal HA binding (set as 100%) in the absence of these inhibitors was calculated as the MFI of HA binding in the treated cultures divided by that of the control cultures × 100 − 100%. For azide, the mean was 130% ± 25%; for nystatin, the mean was 18% ± 3%; and for cytochalasin B, the mean was 89% ± 15% increase above basal HA binding.** P < .01 as compared with the basal HA binding of untreated HPCs. The pattern of inhibition for all three agents was the same for all 6 patient samples analyzed. Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology

Linda M. Pilarski et al. Blood 1999;93:2918-2927 ©1999 by American Society of Hematology