scholarly journals Identification of Sialyl-Lewis(x)-Interacting Protein on Human Spermatozoa

2021 ◽  
Vol 9 ◽  
Author(s):  
Ying Wang ◽  
Weie Zhao ◽  
Si Mei ◽  
Panyu Chen ◽  
Tsz-Ying Leung ◽  
...  
Keyword(s):  
Acrosome Reaction ◽  
Fertilization Rate ◽  
Human Spermatozoa ◽  
Chromosome 1 ◽  
Sialyl Lewis X ◽  
Spectrometric Analysis ◽  
Lewis X ◽  
Interacting Protein ◽  
Sialyl Lewis

Capacitated spermatozoa initiate fertilization by binding to the zona pellucida (ZP). Defective spermatozoa-ZP binding causes infertility. The sialyl-Lewis(x) (SLeX) sequence is the most abundant terminal sequence on the glycans of human ZP glycoproteins involving in spermatozoa-ZP binding. This study aimed to identify and characterize the SLeX-binding proteins on human spermatozoa. By using affinity chromatography followed by mass spectrometric analysis, chromosome 1 open reading frame 56 (C1orf56) was identified to be a SLeX-binding protein of capacitated spermatozoa. The acrosomal region of spermatozoa possessed C1orf56 immunoreactive signals with intensities that increased after capacitation indicating translocation of C1orf56 to the cell surface during capacitation. Treatment with antibody against C1orf56 inhibited spermatozoa-ZP binding and ZP-induced acrosome reaction. Purified C1orf56 from capacitated spermatozoa bound to human ZP. A pilot clinical study was conducted and found no association between the percentage of capacitated spermatozoa with C1orf56 expression and in vitro fertilization (IVF) rate in assisted reproduction treatment. However, the percentage of C1orf56 positive spermatozoa in the acrosome-reacted population was significantly (P < 0.05) lower in cycles with a fertilization rate < 60% when compared to those with a higher fertilization rate, suggesting that C1orf56 may have functions after ZP-binding and acrosome reaction. A larger clinical trial is needed to determine the possible use of sperm C1orf56 content for the prediction of fertilization potential of sperm samples.

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 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 Effect of sialyl Lewis X-glycoliposomes on the inhibition of E-selectin-mediated tumour cell adhesion in vitro

2004 ◽  
Vol 1660 (1-2) ◽  
pp. 31-40 ◽  
Author(s):  
Reinhard Zeisig ◽  
Renate Stahn ◽  
Katrin Wenzel ◽  
Diana Behrens ◽  
Iduna Fichtner
Keyword(s):  
Cell Adhesion ◽  
Tumour Cell ◽  
Sialyl Lewis X ◽  
Lewis X ◽  
Sialyl Lewis

scholarly journals De novo expression of endothelial sialyl Lewis(a) and sialyl Lewis(x) during cardiac transplant rejection: superior capacity of a tetravalent sialyl Lewis(x) oligosaccharide in inhibiting L-selectin-dependent lymphocyte adhesion.

1995 ◽  
Vol 182 (4) ◽  
pp. 1133-1141 ◽  
Author(s):  
J P Turunen ◽  
M L Majuri ◽  
A Seppo ◽  
S Tiisala ◽  
T Paavonen ◽  
...  
Keyword(s):  
De Novo ◽  
Transplant Rejection ◽  
Sialyl Lewis X ◽  
Cardiac Transplant ◽  
Lewis X ◽  
Lymphocyte Adhesion ◽  
Sialyl Lewis ◽  
Sialyl Lewis A ◽  
Lewis A

Acute organ transplant rejection is characterized by a heavy lymphocyte infiltration. We have previously shown that alterations in the graft endothelium lead to increased lymphocyte traffic into the graft. Here, we demonstrate that lymphocytes adhere to the endothelium of rejecting cardiac transplants, but not to the endothelium of syngeneic grafts or normal hearts analyzed with the in vitro Stamper-Woodruff binding assay. Concomitant with the enhanced lymphocyte adhesion, the cardiac endothelium begins to de novo express sialyl Lewis(a) and sialyl Lewis(x) (sLea and sLex) epitopes, which have been shown to be sequences of L-selectin counterreceptors. The endothelium of allografts, but not that of syngeneic grafts or normal controls, also reacted with the L-selectin-immunoglobulin G fusion protein, giving further proof of inducible L-selectin counterreceptors. The lymphocyte adhesion to endothelium could be significantly decreased either by treating the lymphocytes with anti-L-selectin antibody HRL-1, or by treating the tissue sections with sialidase or anti-sLea or anti-sLex monoclonal antibodies. Finally, we synthetized enzymatically several members of the sLex family oligosaccharides and analyzed their ability to block lymphocyte adhesion to cardiac endothelium. The monovalent sLex (a tetramer), divalent sLex (a decamer), and tetravalent sLex (a 22-mer) could all significantly reduce lymphocyte binding, but the inhibition by the tetravalent sLex-construct was clearly superior to other members of the sLex family. The crucial control oligosaccharides, sialyl lactosamines lacking fucose but being otherwise similar to the members of sLex family, had no effect on lymphocyte binding.

scholarly journals N-glycans of core2 β(1,6)-N-acetylglucosaminyltransferase-I (C2GnT-I) but not those of α(1,3)-fucosyltransferase-VII (FucT-VII) are required for the synthesis of functional P-selectin glycoprotein ligand-1 (PSGL-1): effects on P-, L- and E-selectin binding

2005 ◽  
Vol 391 (3) ◽  
pp. 491-502 ◽  
Author(s):  
Maëlle Prorok-Hamon ◽  
Frédéric Notel ◽  
Sylvie Mathieu ◽  
Claire Langlet ◽  
Minoru Fukuda ◽  
...  
Keyword(s):  
Drug Targets ◽  
Sialyl Lewis X ◽  
Lewis X ◽  
Sialyl Lewis ◽  
Selectin Ligands ◽  
Glycosylation Sites ◽  
Glycan Chain ◽  
Golgi Compartment ◽  
Selectin Binding

C2GnT-I [core2 β(1,6)-N-acetyglucosaminyltransferase-I] and FucT-VII [α(1,3)-fucosyltransferase-VII] are the key enzymes for the biosynthesis of sialyl-Lewis x determinants on selectin ligands and therefore they represent good drug targets for the treatment of inflammatory disorders and other pathologies involving selectins. In the present study, we examined the importance of N-glycosylation for the ability of C2GnT-I and FucT-VII to generate functional selectin ligands, particularly the PSGL-1 (P-selectin glycoprotein ligand-1). We found that (i) both enzymes have their two N-glycosylation sites occupied, (ii) for C2GnT-I, the N-glycan chain linked to Asn-95 significantly contributes to the synthesis of functional PSGL-1 and is required to localize the enzyme to the cis/medial-Golgi compartment, (iii) all N-glycosylation-deficient proteins of FucT-VII displayr a dramatic impairment of their in vitro enzymatic activities, but retain their ability to fucosylate the core2-modified PSGL-I and to generate P- and L-selectin binding, and (iv) the glycomutants of FucT-VII fail to synthesize sialyl-Lewis x or to generate E-selectin binding unless core2-modified PSGL-1 is present. All combined, our results show a differential functional impact of N-glycosylation on C2GnT-1 and FucT-VII and disclose that a strongly reduced FucT-VII activity retains the ability to fucosylate PSGL-1 on the core2-based binding site(s) for the three selectins.

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.

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