scholarly journals Fluorinated per-acetylated GalNAc metabolically alters glycan structures on leukocyte PSGL-1 and reduces cell binding to selectins

Blood ◽  
2010 ◽  
Vol 115 (6) ◽  
pp. 1303-1312 ◽  
Author(s):  
Dhananjay D. Marathe ◽  
Alexander Buffone ◽  
E. V. Chandrasekaran ◽  
Jun Xue ◽  
Robert D. Locke ◽  
...  
Keyword(s):  
Inflammatory Diseases ◽  
Leukocyte Adhesion ◽  
Human Leukocyte ◽  
Sialyl Lewis X ◽  
Metabolic Inhibitor ◽  
Growth Media ◽  
Cell Binding ◽  
Lewis X ◽  
Sialyl Lewis

Abstract Novel strategies to control the binding of adhesion molecules belonging to the selectin family are required for the treatment of inflammatory diseases. We tested the possibility that synthetic monosaccharide analogs can compete with naturally occurring sugars to alter the O-glycan content on human leukocyte cell surface selectin-ligand, P-selectin glycoprotein ligand-1 (PSGL-1). Resulting reduction in the sialyl Lewis-X–bearing epitopes on this ligand may reduce cell adhesion. Consistent with this hypothesis, 50μM per-acetylated 4F-GalNAc added to the growth media of promyelocytic HL-60 cells reduced the expression of the cutaneous lymphocyte associated-antigen (HECA-452 epitope) by 82% within 2 cell doubling cycles. Cell binding to all 3 selectins (L-, E-, and P-selectin) was reduced in vitro. 4F-GalNAc was metabolically incorporated into PSGL-1, and this was accompanied by an approximately 20% reduction in PSGL-1 glycan content. A 70% to 85% reduction in HECA-452 binding epitope and N-acetyl lactosamine content in PSGL-1 was also noted on 4F-GalNAc addition. Intravenous 4F-GalNAc infusion reduced leukocyte migration to the peritoneum in a murine model of thioglycolate-induced peritonitis. Thus, the compound has pharmacologic activity. Overall, the data suggest that 4F-GalNAc may be applied as a metabolic inhibitor to reduce O-linked glycosylation, sialyl Lewis-X formation, and leukocyte adhesion via the selectins.

Glycan engineering using synthetic ManNAc analogues enhances sialyl-Lewis-X expression and adhesion of leukocytes

2021 ◽  
Author(s):  
Anam Tasneem ◽  
Shubham Parashar ◽  
Tanya Jain ◽  
Simran Aittan ◽  
Jyoti Rautela ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Inflammatory Diseases ◽  
Leukocyte Adhesion ◽  
Fold Increase ◽  
Sialyl Lewis X ◽  
Lewis X ◽  
Sialyl Lewis ◽  
Acute Myeloid Leukemia Cells ◽  
Coated Surfaces ◽  
Cell Cell

Cell surface glycans, depending on their structures and dynamic modifications, act as the first point of contact and regulate cell-cell, cell-matrix, and cell-pathogen interactions. Particularly, the sialyl-Lewis-X (sLeX, CD15s) tetrasaccharide epitope, expressed on both glycoproteins and gangliosides, participates in leukocyte extravasation via interactions with selectins expressed on endothelial cells, lymphocytes, and platelets (CD62-E/L/P). Neutrophils carrying sLeX epitopes are thought to be responsible for chronic inflammatory diseases resulting in plaque formation and atherosclerosis. Intense efforts have been devoted to the development of sLeX mimetics for inhibition of cell adhesion. On the other hand, dysregulated expression of sLeX and poor extravasation are the major underlying causes of leukocyte adhesion deficiency-II (LAD-II) disorders that result in frequent infections and poor immune response. We hypothesized that metabolic processing of peracetyl N-(cycloalkyl)acyl-D-mannosamine derivatives, through the sialic acid pathway, might result in the expression of sialoglycans with altered hydrophobicity which in-turn could modulate their binding to endogenous lectins, including selectins. Herein, we show that treatment of HL-60 (human acute myeloid leukemia) cells with peracetyl N-cyclobutanoyl-D-mannosamine (Ac4ManNCb), at 50 microM for 48 h, resulted in a robust three to four fold increase in the binding of anti-sLeX (CSLEX1) antibody and enhanced cell adhesion to E-selectin coated surfaces; while the corresponding straight-chain analogue, peracetyl N-pentanoyl-D-mannosamine (Ac4ManNPent), and peracetyl N-cyclopropanoyl-D-mannosamine (Ac4ManNCp) both resulted in 2.0-2.5fold increase compared to controls. The ability to enhance sLeX expression using small molecules has the potential to provide novel opportunities to address challenges in the treatment of immune deficiency disorders.

In Vitro Selection of the RNA Aptamer against the Sialyl Lewis X and Its Inhibition of the Cell Adhesion

2001 ◽  
Vol 281 (1) ◽  
pp. 237-243 ◽  
Author(s):  
Sunjoo Jeong ◽  
Tae-Yeon Eom ◽  
Se-Jin Kim ◽  
Seong-Wook Lee ◽  
Jaehoon Yu
Keyword(s):  
Cell Adhesion ◽  
In Vitro Selection ◽  
Sialyl Lewis X ◽  
Rna Aptamer ◽  
Lewis X ◽  
Sialyl Lewis ◽  
Selection Of

scholarly journals The crystal structure of staphylococcal superantigen-like protein 11 in complex with sialyl Lewis X reveals the mechanism for cell binding and immune inhibition

2007 ◽  
Vol 66 (6) ◽  
pp. 1342-1355 ◽  
Author(s):  
Matthew C. Chung ◽  
Bruce D. Wines ◽  
Heather Baker ◽  
Ries J. Langley ◽  
Edward N. Baker ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Sialyl Lewis X ◽  
Cell Binding ◽  
Lewis X ◽  
Sialyl Lewis

scholarly journals Mechanical Shedding of L-selectin from the Neutrophil Surface during Rolling on Sialyl Lewis x under Flow

2006 ◽  
Vol 282 (7) ◽  
pp. 4812-4820 ◽  
Author(s):  
Dooyoung Lee ◽  
Joanne B. Schultz ◽  
Philip A. Knauf ◽  
Michael R. King
Keyword(s):  
Flow Chamber ◽  
Mechanical Force ◽  
Mitogen Activated Protein Kinase ◽  
Sialyl Lewis X ◽  
Lewis X ◽  
Rolling Velocity ◽  
Sialyl Lewis ◽  
Mitogen Activated Protein ◽  
Neutrophil Surface

The interaction of L-selectin expressed on leukocytes with endothelial cells leads to capture and rolling and is critical for the recruitment of leukocytes into sites of inflammation. It is known that leukocyte activation by chemoattractants, the change of osmotic pressure in cell media, or cross-linking of L-selectin all result in rapid shedding of L-selectin. Here we present a novel mechanism for surface cleavage of L-selectin on neutrophils during rolling on a sialyl Lewis x-coated surface that involves mechanical force. Flow cytometry and rolling of neutrophils labeled with Qdot®-L-selectin antibodies in an in vitro flow chamber showed that the mechanical shedding of L-selectin occurs during rolling and depends on the amount of shear applied. In addition, the mechanical L-selectin shedding causes an increase in cell rolling velocity with rolling duration, suggesting a gradual loss of L-selectin and is mediated by p38 mitogen-activated protein kinase activation. Thus, these data show that mechanical force induces the cleavage of L-selectin from the neutrophil surface during rolling and therefore decreases the adhesion of cells to a ligand-presenting surface in flow.

scholarly journals Evaluation of Strategies Aimed at Improving Liver Progenitor Cell Rolling and Subsequent Adhesion to the Endothelium

2020 ◽  
Vol 29 ◽  
pp. 096368972091270
Author(s):  
Pierre Edouard Dollet ◽  
Mei Ju Hsu ◽  
Jérôme Ambroise ◽  
Milena Rozzi ◽  
Joachim Ravau ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Alternative Methods ◽  
Sialyl Lewis X ◽  
Thermosensitive Polymer ◽  
Lewis X ◽  
Promising Alternative ◽  
Sialyl Lewis ◽  
Selectin Ligand

Adult-derived human liver stem/progenitor cells (ADHLSCs) are a promising alternative to orthotopic liver transplantation in the treatment of inborn errors of metabolism. However, as is the case with many mesenchymal stromal cells, ADHLSCs have shown a low level of engraftment, which could be explained by the fact that they lack expression of selectin ligand and LFA-1 and only slightly express VLA- 4, molecules that have been shown to be involved in cell adhesion to the endothelium. In this paper, we have investigated strategies to increase their rolling and adhesion during the homing process by (1) adding a selectin ligand (Sialyl Lewis X) to their surface using biotinyl- N-hydroxy-succinimide–streptavidin bridges, and (2) protecting the adhesion proteins from trypsinization-induced damage using a thermosensitive polymer for cell culture and a nonenzymatic cell dissociation solution (CDS) for harvest. Despite increasing adhesion of ADHLSCs to E-selectin during an adhesion test in vitro performed under shear stress, the addition of Sialyl Lewis X did not increase adhesion to endothelial cells under the same conditions. Cultivating cells on a thermosensitive polymer and harvesting them with CDS increased their adhesion to endothelial cells under noninflammatory conditions, compared to the use of trypsin. However, we were not able to demonstrate any improvement in cell adhesion to the endothelium following culture on polymer and harvest with CDS, suggesting that alternative methods of improving engraftment still need to be evaluated.

Recognition of consensus CHO structure in ligands for selectins by novel antibody against sialyl Lewis X

1995 ◽  
Vol 269 (4) ◽  
pp. H1282-H1287 ◽  
Author(s):  
T. Tamatani ◽  
M. Suematsu ◽  
K. Tezuka ◽  
N. Hanzawa ◽  
T. Tsuji ◽  
...  
Keyword(s):  
Sialyl Lewis X ◽  
Consensus Structure ◽  
Carbohydrate Structure ◽  
Lewis X ◽  
High Endothelial Venules ◽  
Cos Cells ◽  
Cell Lysate ◽  
Sialyl Lewis

The selectins (L, E, and P) play an important role in the earliest events of the inflammatory response, leading to the “rolling” phenomenon. All selectins react with sialyl Lewis X (SLex) in vitro, possibly suggesting that their ligands have a consensus structure. 2H5 is a monoclonal antibody against SLex that blocks L-selectin-mediated adhesion. 2H5 inhibited adhesion of HL-60 cells to P- and E-selectin-producing COS cells in vitro and immunoprecipitated a P-selectin glycoprotein ligand-1-like glycoprotein from HL-60 cell lysate, suggesting that it recognizes a functional consensus structure on the ligands for all selectins. 2H5 reacted not only with human but also with rat and mouse neutrophils. 2H5 is the first antibody against SLex that recognizes neutrophils of nonhuman mammals. The carbohydrate structure recognized by 2H5 was present not only on high endothelial venules of rat lymphoid organs but also on the endothelial cells of nonlymphoid organs. Furthermore, administration of the antibody markedly inhibited L- and P-selectin-mediated neutrophil rolling and adhesion in rat mesenteric venules in vivo. These results provide evidence for the presence of a consensus carbohydrate structure on the ligands for all selectins. The consensus structure thus has the potential to serve as a therapeutic target.

scholarly journals The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide

1992 ◽  
Vol 117 (4) ◽  
pp. 895-902 ◽  
Author(s):  
C Foxall ◽  
SR Watson ◽  
D Dowbenko ◽  
C Fennie ◽  
LA Lasky ◽  
...  
Keyword(s):  
Adhesion Molecule ◽  
Leukocyte Adhesion ◽  
Structural Features ◽  
Sialyl Lewis X ◽  
Lewis X ◽  
Leukocyte Adhesion Molecule ◽  
Lectin Domain ◽  
Calcium Dependent ◽  
Carbohydrate Epitope ◽  
Sialyl Lewis

The selectins (lectin-EGF-complement binding-cell adhesion molecules [LEC-CAMs]) are a family of mammalian receptors implicated in the initial interactions between leukocytes and vascular endothelia, leading to lymphocyte homing, platelet binding, and neutrophil extravasation. The three known selectins, L-selectin (leukocyte adhesion molecule-1 [LECAM-1]), E-selectin (endothelial-leukocyte adhesion molecule-1 [ELAM-1]), and P-selectin (GMP-140) share structural features that include a calcium-dependent lectin domain. The sialyl Lewis(x) carbohydrate epitope has been reported as a ligand for both E- and P-selectins. Although L-selectin has been demonstrated to bind to carbohydrates, structural features of potential mammalian carbohydrate ligand(s) have not been well defined. Using an ELISA developed with a sialyl Lewis(x)-containing glycolipid and an E-selectin-IgG chimera, we have demonstrated the direct binding of the L-selectin-IgG chimera to sialyl Lewis(x). This recognition was calcium dependent, and could be blocked by Mel-14 antibody but not by other antibodies. Recognition was confirmed by the ability of cells expressing the native L-selectin to adhere to immobilized sialyl Lewis(x). These data suggest that the sialyl Lewis(x) oligosaccharide may form the basis of a recognition domain common to all three selectins.

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