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Sialyl Lewis X moiety on rat polymorphonuclear leukocytes responsible for binding to rat E-selectin
E Misugi1, N Kawamura, N Imanishi
1Research Center, Sumitomo Pharmaceuticals Co., Ltd., Osaka, Japan.
Abstract:
We investigated the presence of the carbohydrate ligand for E-selectin on the cell surface of rat polymorphonuclear leukocytes (PMN). Rat PMN, isolated from peripheral blood, adhered to recombinant rat E-selectin-coated microplates. The adhesion was inhibited either by an anti-rat E-selectin monoclonal antibody (MAb), a sialyl Lewis X (SLex) oligosaccharide or neuraminidase digestion. FACS analysis revealed the expression of SLex as well as other adhesion molecules such as L-selectin, CD11a, CD11b and CD18 on the cell surface of rat PMN. The binding of an anti-SLex MAb KM93 to rat PMN was inhibited competitively by a SLex but not by a Lewis X (Lex). The reactivities of two anti-SLex MAbs, KM93 and CSLEX-1, or an anti-Lex MAb BC90/45 to rat PMN differed from those to human PMN. These results suggest that rat PMN contain the SLex-moiety, which binds to rat E-selectin and is different from that of human PMN.
Insights
Rat polymorphonuclear leukocytes (PMN) express sialyl Lewis X (SLex) on their surface, which binds to E-selectin. This interaction is crucial for cell adhesion and differs from human PMN.
Area of Science:
- Immunology
- Cell Biology
- Glycobiology
Background:
- E-selectin is a cell adhesion molecule involved in inflammatory responses.
- Polymorphonuclear leukocytes (PMN) play a key role in innate immunity and inflammation.
- The carbohydrate ligand for E-selectin, sialyl Lewis X (SLex), mediates cell adhesion.
Purpose of the Study:
- To investigate the presence and characteristics of the E-selectin ligand on rat PMN.
- To determine if rat PMN express SLex and its role in E-selectin-mediated adhesion.
- To compare the E-selectin ligand on rat PMN with that of human PMN.
Main Methods:
- Isolation of rat peripheral blood polymorphonuclear leukocytes (PMN).
- E-selectin-mediated PMN adhesion assays using coated microplates.
- Inhibition studies using anti-E-selectin monoclonal antibody (MAb), sialyl Lewis X (SLex) oligosaccharide, and neuraminidase.
- Flow cytometry (FACS) analysis for cell surface molecule expression.
- Competitive binding assays with anti-SLex MAbs.
Main Results:
- Rat PMN adhered to E-selectin-coated plates, and this adhesion was inhibited by anti-E-selectin MAb, SLex, and neuraminidase.
- FACS analysis confirmed SLex expression on rat PMN, along with other adhesion molecules (L-selectin, CD11a, CD11b, CD18).
- Binding of anti-SLex MAbs to rat PMN showed differences compared to human PMN, indicating distinct SLex structures.
Conclusions:
- Rat PMN possess the sialyl Lewis X (SLex) moiety that binds to rat E-selectin.
- The SLex-moiety on rat PMN involved in E-selectin binding is structurally distinct from that found on human PMN.
- These findings highlight species-specific differences in E-selectin ligand expression and function.