Discordant expression of selectin ligands and sialyl Lewis x–related epitopes on murine myeloid cells

Kobzdej, M; Leppänen, Anne; Ramachandran, V.; Cummings, Richard D.; McEver, Rodger P.

doi:10.1182/blood-2002-06-1799

Cited by 53 publications

(54 citation statements)

References 55 publications

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“…The late myeloblast cell line HL-60 also expresses E-and P-selectin ligands in a uniform manner [13]. We found that the levels of reactivity by mouse, human, and rat E-selectin/Fc with these cells were similar to neutrophils (Fig.…”

Section: The Binding Activity By Different E-selectin/fc Constructs Imentioning

confidence: 56%

Varied levels of reactivity by different E-selectin/Fc constructs with cutaneous lymphocyte-associated antigen (CLA)+ CD4+ T cells

Walcheck

2007

Immunology Letters

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SUMMARYT cells utilize the vascular adhesion molecule E-selectin to enter inflamed skin. T lymphocytes identified by the mAb HECA-452 [cutaneous lymphocyte-associated antigen (CLA) T cells] are enriched in E-selectin ligand expressing cells. However, the proportion of CLA + T cells reactive with an E-selectin/Fc chimeric construct, as determined by flow cytometry, can vary considerably between studies. One important variable in these studies has been the E-selectin/Fc chimeric used to assess ligand expression. We therefore compared the reactivity of mouse, rat, and human E-selectin/Fc from the same widely used commercial source with peripheral blood CLA + CD4 + T cells, neutrophils, and the promyelocytic cell line HL-60 by flow cytometry and by shear flow assays. We observed that the binding activities of the different E-selectin/Fc chimeras were considerably different. Mouse E-selectin/Fc demonstrated the highest binding activity with human neutrophils and HL-60 cells by both assay approaches, whereas human E-selectin/Fc demonstrated the lowest binding activity. In addition, mouse E-selectin/Fc binding increased essentially in a linear manner with increasing expression of CLA by CD4 + T cells, whereas human and rat E-selectin/Fc binding occurred with only a subset of CLA + CD4 + T cells. We conclude that there is substantial variability in the reactivity of different E-selectin/Fc constructs, thus caution should be used when assessing E-selectin ligand expression with these reagents. For instance, the discordance in expression of CLA and E-selectin ligands by T cells may in part be due to the E-selectin/Fc construct being used.

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Section: The Binding Activity By Different E-selectin/fc Constructs Imentioning

confidence: 56%

Varied levels of reactivity by different E-selectin/Fc constructs with cutaneous lymphocyte-associated antigen (CLA)+ CD4+ T cells

Walcheck

2007

Immunology Letters

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“…The apparent importance of FUT9 in human leukocyte adhesion is not entirely unexpected because it is well recognized that the E-selectin ligands in human leukocytes differ from those in the mouse (1,5). In particular, E-selectin ligands on mouse but not human neutrophils are pronase-sensitive (7,8). 3 In this regard, Nimrichter et al (47) suggest that a relatively minor monosialylated, polyfucosylated glycolipid with poly-lactosamine units NeuAc␣␣2,3Gal␤1,4GlcNAc␤1,3[Gal␤1,4 (Fuc␣1,3)GlcNAc␤1,3] 2 [Gal␤1,4GlcNAc␤1,3] 2 Gal␤1,4Glc␤Cer…”

Section: Fut7mentioning

confidence: 99%

“…In this regard, although E-selectin binding to mouse granulocytes is completely lost upon pronase digestion, it is only partially reduced in human cells (7,8).…”

mentioning

confidence: 99%

Silencing α1,3-Fucosyltransferases in Human Leukocytes Reveals a Role for FUT9 Enzyme during E-selectin-mediated Cell Adhesion

Buffone

Mondal

Gupta

et al. 2013

Journal of Biological Chemistry

106

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Background: During inflammation, the selectins engage glycosylated macromolecules expressed on blood leukocytes under fluid shear conditions. Results: Although all three myeloid ␣1,3-fucosyltransferases FUT9, FUT7, and FUT4 regulate human E-selectin ligand biosynthesis, FUT7 and FUT4 are sufficient to form L/P-selectin ligands. Conclusion: FUT9 plays a significant role during human, but not mouse, leukocyte-endothelial interactions. Significance: This study identifies potential ␣(1,3)FUTs regulating inflammation in humans.

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“…7,8 With regard to E-selectin, however, human but not mouse granulocyte rolling on this selectin is insensitive to pronase treatment. 9,10 Thus, protease-insensitive gangliosides may be physiological E-selectin ligands that are unique to humans. 11 Among the a1,3fucosyltransferases, the enzyme FUT9 reportedly plays a more significant role during human leukocyte adhesion, compared with FUT7 and FUT4 which are the dominant players in mice.…”

Section: Introductionmentioning

confidence: 99%

ST3Gal-4 is the primary sialyltransferase regulating the synthesis of E-, P-, and L-selectin ligands on human myeloid leukocytes

Mondal

Buffone

Stolfa

et al. 2015

Blood

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• A single a(2,3) sialyltransferase, ST3Gal-4, controls sLe X biosynthesis on N-and O-glycans in cells of human myeloid lineage.• Blocking this enzyme activity prevents human neutrophil adhesion to E-, P-, and L-selectin.The precise glycosyltransferase enzymes that mediate selectin-ligand biosynthesis in human leukocytes are unknown. This knowledge is important because selectin-mediated cell tethering and rolling is a critical component of both normal immune response and various vascular disorders. We evaluated the role of 3 a(2,3)sialyltransferases, ST3Gal-3, -4, and -6, which act on the type II N-Acetyllactosamine structure (Galb1,4GlcNAc) to create sialyl Lewis-X (sLe X ) and related sialofucosylated glycans on human leukocytes of myeloid lineage. These genes were either silenced using lentiviral short hairpin RNA (shRNA) or functionally ablated using the clustered regularly interspaced short palindromic repeat/ Cas9 technology. The results show that ST3Gal-4, but not ST3Gal-3 or -6, is the major sialyltransferase regulating the biosynthesis of E-, P-, and L-selectin ligands in humans. Reduction in ST3Gal-4 activity lowered cell-surface HECA-452 epitope expression by 75% to 95%. Glycomics profiling of knockouts demonstrate an almost complete loss of the sLe X epitope on both leukocyte N-and O-glycans. In cell-adhesion studies, ST3Gal-4 knockdown/knockout cells displayed 90% to 100% reduction in tethering and rolling density on all selectins. ST3Gal-4 silencing in neutrophils derived from human CD34 1 hematopoietic stem cells also resulted in 80% to 90% reduction in cell adhesion to all selectins. Overall, a single sialyltransferase regulates selectin-ligand biosynthesis in human leukocytes, unlike mice where multiple enzymes contribute to this function. (Blood. 2015;125(4):687-696)

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Discordant expression of selectin ligands and sialyl Lewis x–related epitopes on murine myeloid cells

Cited by 53 publications

References 55 publications

Varied levels of reactivity by different E-selectin/Fc constructs with cutaneous lymphocyte-associated antigen (CLA)+ CD4+ T cells

Varied levels of reactivity by different E-selectin/Fc constructs with cutaneous lymphocyte-associated antigen (CLA)+ CD4+ T cells

Silencing α1,3-Fucosyltransferases in Human Leukocytes Reveals a Role for FUT9 Enzyme during E-selectin-mediated Cell Adhesion

ST3Gal-4 is the primary sialyltransferase regulating the synthesis of E-, P-, and L-selectin ligands on human myeloid leukocytes

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