Suppression of allogeneic reactivity in vitro by the syncytiotrophoblast membrane glycocalyx of the human term placenta is carbohydrate dependent.
Immunosuppressive factors isolated from trophoblast are known to block both innate and major histocompatibility complex (MHC)-dependent cell-mediated immune responses in vitro and, in some cases, in vivo. We investigated the biochemical nature of these factors, which is presently unknown. Immunosuppressive activity, assessed by inhibition of two-way MLR, was extracted from term syncytiotrophoblast microvilli using 3 M KCl. The activity resisted both extensive pronase digestion and heating to 90 degrees C for 1 h, demonstrating that intact membrane proteins were not required. Although purified protein-linked oligosaccharides released by hydrazinolysis from the syncytiotrophoblast membrane were themselves inactive, they blocked the immunosuppressive activity of the KCl extract. After pronase digestion, the activity could be fractionated by TSK 55S gel filtration, followed by C18 reverse-phase chromatography. Sequential exoglycosidase digestion of hydrazine-released sugars of the active fraction demonstrated that it contained neutral N-linked oligomannose and hybrid oligosaccharides, which normally make up < 3% of the total syncytiotrophoblast-derived protein glycan library. These glycopeptides of the active fraction were associated with membrane phospholipid micelles. The possible mechanism by which incompletely processed N-linked oligosaccharides expressed by a variety of syncytiotrophoblast membrane glycoproteins may block allogeneic reactivity when presented as polyvalent sugar groups is discussed.
Arkwright
P D
PD
Department of Biochemistry, University of Oxford, UK.
Rademacher
T W
TW
Boutignon
F
F
Dwek
R A
RA
Redman
C W
CW
D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000276:immunology*; D002455:Cell Division; D005260:Female; D006023:Glycoproteins / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007108:Immune Tolerance / Q000502:physiology*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000378:metabolism; D008563:Membrane Lipids / Q000276:immunology; D008871:Microvilli / Q000276:immunology / Q000378:metabolism; D011134:Polysaccharides / Q000276:immunology* / Q000378:metabolism; D011247:Pregnancy; D011402:Pronase; D014327:Trophoblasts / Q000276:immunology* / Q000378:metabolism
D002241:Carbohydrates; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D008563:Membrane Lipids; D011134:Polysaccharides; D011402:Pronase