Analysis of glycoprotein E-selectin ligANDs on human and mouse marrow cells enriched for hematopoietic stem/progenitor cells
Burdick, Monica M.
Dagia, Nilesh M.
Chu, Julia T.
Fuhlbrigge, Robert C.
Sackstein, Robert D.
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Online Publication Date2011-06-09
Print Publication Date2011-08-18
Permanent link to this recordhttp://hdl.handle.net/10754/565953
MetadataShow full item record
AbstractAlthough well recognized that expression of E-selectin on marrow microvessels mediates osteotropism of hematopoietic stem/progenitor cells (HSPCs), our knowledge regarding the cognate E-selectin ligand(s) on HSPCs is incomplete. Flow cytometry using E-selectin-Ig chimera (E-Ig) shows that human marrow cells enriched for HSPCs (CD34+ cells) display greater E-selectin binding than those obtained from mouse (lin-/Sca-1+/c-kit+ [LSK] cells). To define the relevant glycoprotein E-selectin ligands, lysates from human CD34+ and KG1a cells and from mouse LSK cells were immunoprecipitated using E-Ig and resolved byWestern blot using E-Ig. In both human and mouse cells, E-selectin ligand reactivity was observed at ∼ 120- to 130-kDa region, which contained two E-selectin ligands, the P-selectin glycoprotein ligand- 1 glycoform "CLA," and CD43. Human, but not mouse, cells displayed a prominent ∼ 100-kDa band, exclusively comprising the CD44 glycoform "HCELL."E-Ig reactivity was most prominent on CLA in mouse cells and on HCELL in human cells. To further assess HCELL's contribution to E-selectin adherence, complementary studies were performed to silence (via CD44 siRNA) or enforce its expression (via exoglycosylation). Under physiologic shear conditions, CD44/HCELL-silenced human cells showed striking decreases (> 50%) in E-selectin binding. Conversely, enforced HCELL expression of LSK cells profoundly increased E-selectin adherence, yielding > 3-fold more marrow homing in vivo. These data define the key glycoprotein E-selectin ligands of human and mouse HSPCs, unveiling critical species-intrinsic differences in both the identity and activity of these structures. © 2011 by The American Society of Hematology.
PublisherAmerican Society of Hematology
PubMed Central IDPMC3158712
- The biology of CD44 and HCELL in hematopoiesis: the 'step 2-bypass pathway' and other emerging perspectives.
- Authors: Sackstein R
- Issue date: 2011 Jul
- differential L-selectin binding activities of human hematopoietic cell L-selectin ligands, HCELL and PSGL-1.
- Authors: Dimitroff CJ, Lee JY, Schor KS, Sandmaier BM, Sackstein R
- Issue date: 2001 Dec 14
- HCELL is the major E- and L-selectin ligand expressed on LS174T colon carcinoma cells.
- Authors: Burdick MM, Chu JT, Godar S, Sackstein R
- Issue date: 2006 May 19
- Ex vivo glycan engineering of CD44 programs human multipotent mesenchymal stromal cell trafficking to bone.
- Authors: Sackstein R, Merzaban JS, Cain DW, Dagia NM, Spencer JA, Lin CP, Wohlgemuth R
- Issue date: 2008 Feb
- CD44 on LS174T colon carcinoma cells possesses E-selectin ligand activity.
- Authors: Hanley WD, Burdick MM, Konstantopoulos K, Sackstein R
- Issue date: 2005 Jul 1