Purification and characterization of an immunomodulatory endometrial protein, glycodelin.
Human glycodelin is synthesized by endometrial cells in the late secretory phase and early pregnancy under hormonal regulation. Whereas the precise physiological functions of glycodelin are unknown, its expression during embryonic nidation and its inhibition of T cell proliferation suggest an immunomodulatory role. We purified human glycodelin from first trimester human decidual cytosol by using a rapid two-step high-performance liquid chromatography method and investigated its effects on human monocyte migration. Human U937 cells were used as a model of monocyte chemotaxis in Boyden chamber migration assays. N-Formyl-Met-Leu-Phe and the beta-chemokine RANTES (regulated on activation normal T cell expressed and secreted) were used as monocyte chemoattractants. Purified glycodelin inhibited monocyte migration in a dose-dependent fashion (IC50 = 550 nm). Glycodelin activity was totally reversed by heat inactivation (95 degrees C x 15 min) and neutralized by pretreatment with specific anti-glycodelin antibodies. Deglycosylated glycodelin was equipotent to intact glycodelin in the monocyte migration assay. 125I-Glycodelin binding to whole U937 cells revealed a single, saturable site with a Kd = 48 +/- 21 nm by Scatchard analysis. Cross-linking studies indicated that glycodelin binds to a high molecular mass (approximately 250 kDa) protein complex at the monocyte cell surface. Our findings support the hypothesis that glycodelin reduces the local maternal inflammatory response toward the implantation of a semiallogeneic conceptus.
Vigne
J L
JL
Center for Reproductive Sciences, Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, California 94143-0556, USA.
Hornung
D
D
Mueller
M D
MD
Taylor
R N
RN
D000595:Amino Acid Sequence; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D018946:Chemokine CCL5 / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000502:physiology; D003656:Decidua / Q000737:chemistry / Q000276:immunology*; D004717:Endometrium / Q000737:chemistry / Q000276:immunology; D005260:Female; D000071816:Glycodelin; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D006031:Glycosylation; D006801:Humans; D007166:Immunosuppressive Agents / Q000737:chemistry / Q000302:isolation & purification / Q000493:pharmacokinetics / Q000494:pharmacology*; D007457:Iodine Radioisotopes; D008213:Lymphocyte Activation / Q000187:drug effects; D008969:Molecular Sequence Data; D009000:Monocytes / Q000187:drug effects / Q000502:physiology; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D011261:Pregnancy Trimester, First; D013601:T-Lymphocytes / Q000276:immunology
D018946:Chemokine CCL5; D000071816:Glycodelin; D006023:Glycoproteins; D007166:Immunosuppressive Agents; D007457:Iodine Radioisotopes; C045246:PAEP protein, human; D011257:Pregnancy Proteins