A (1-->3)-beta-D-linked heptasaccharide is the unit ligand for glucan pattern recognition receptors on human monocytes.
Glucans are fungal cell wall polysaccharides which stimulate innate immune responses. We determined the minimum unit ligand that would bind to glucan receptors on human U937 cells using laminarin-derived pentaose, hexaose, and heptaose glucan polymers. When U937 membranes were pretreated with the oligosaccharides and passed over a glucan surface, only the heptasaccharide inhibited the interaction of glucan with membrane receptors at a K(d) of 31 microM (95% CI 20-48 microM) and 100% inhibition. However, the glucan heptasaccharide did not stimulate U937 monocyte NFkappaB signaling, nor did it increase survival in a murine model of polymicrobial sepsis. Laminarin, a larger and more complex glucan polymer (M(w) = 7700 g/mol), only partially inhibited binding (61 +/- 4%) at a K(d) of 2.6 microM (99% CI 1.7-4.2 microM) with characteristics of a single binding site. These results indicate that a heptasaccharide is the smallest unit ligand recognized by macrophage glucan receptors. The data also indicate the presence of at least two glucan-binding sites on U937 cells and that the binding sites on human monocyte/macrophages can discriminate between glucan polymers. The heptasaccharide and laminarin were receptor antagonists, but they were not receptor agonists with respect to activation of NFkappaB-dependent signaling pathways or protection against experimental sepsis.
Lowe
E
E
Department of Surgery, James H. Quillen College of Medicine, East Tennessee State University, Johnson City 37614-1708, USA.
Rice
P
P
Ha
T
T
Li
C
C
Kelley
J
J
Ensley
H
H
Lopez-Perez
J
J
Kalbfleisch
J
J
Lowman
D
D
Margl
P
P
Browder
W
W
Williams
D
D
D000818:Animals; D001665:Binding Sites; D004305:Dose-Response Relationship, Drug; D005936:Glucans / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008297:Male; D051379:Mice; D008812:Mice, Hairless; D008813:Mice, Inbred ICR; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D011134:Polysaccharides / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding; D011971:Receptors, Immunologic / Q000187:drug effects / Q000378:metabolism*; D018805:Sepsis / Q000276:immunology; D020298:U937 Cells
D005936:Glucans; D008024:Ligands; D016328:NF-kappa B; D011134:Polysaccharides; D011971:Receptors, Immunologic; C045268:beta-glucan receptor; C008247:laminaran