2006 Fiscal Year Final Research Report Summary
Study of anti-laminin-1 autoantibodies in endometriosis-associated infertility
| Project/Area Number |
17591738
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Obstetrics and gynecology
|
|---|
| Research Institution | Okayama University |
|---|
Principal Investigator |
INAGAKI Junko Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, research assistant, 大学院医歯薬学総合研究科, 助手 (90271056)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MATSUURA Eiji Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, associate professor, 大学院医歯薬学総合研究科, 助教授 (20181688)
NAKATSUKA Mikiya Graduate School of Medicine, professor, 医学部, 教授 (40273990)
|
|---|
| Project Period (FY) |
2005 – 2006
|
| Keywords | mouse laminin-1 / anti-laminin-1 antibodies / endometriosis-associated infertility / anti-carbohydrate antibodies / angiogcnesis |
|---|
| Research Abstract |
1. Anti-laminin-1 autoantibodies in infertile patients with endometriosis recognized N-linked oligosaccharide chains of mouse laminin-1. A percentage of patients expressed a natural antibody that recognizes the a-Gal (Galal- 3Ga1β1-4G1cNAc-R) sugar chain epitope. However, the removal of terminal sialic acid residues from mouse laminin-1 resulted in a complete loss of antibody binding by most patients' sera. To identify real autoantigens of these antibodies, lysates from ectopic and eutopic endometrium, and ovarian cyst, peritoneal fluids, and sera from infertile patients with endometriosis or healthy controls were examined by Western blot analysis. Autoantigens were not detected, therefore, we analyze the structure of the oligosaccharide chain epitope using HPLC/MS. Based on the determined epitope structure, we will predict the real autoantigens.
2. We studied the effect of a recently established anti-laminin-1 mouse monoclonal IgM (AK8), an affinity-purified anti-laminin-1 IgG from the
… More
serum of an infertile patient with endometriosis, and a polyclonal anti-laminin rabbit IgG on mouse embryo implantation in vitro. AK8 and the polyclonal anti-laminin rabbit IgG did not have an effect on blastocyst hatching, but strongly inhibited the blastocyst adhesion and outgrowth in about 100 % of the blastocysts. On the other hand, the affinity-purified IgG specific for the oligosaccharide chains of laminin-1 did not affect blastocyst hatching or adhesion, but inhibited the area of outgrowth in about 30 % of the blastocysts.
3. Sera from some infertile patients with endometriosis cross-reacted with the peptide moiety of laminin γl chain. Laminin-1 contains about 20 angiogenic peptide sequences. Among of them, angiogenic peptide sequence of γ1 chain (C16 peptide) binds to integrin α5β1 and αvβ3 of endothelial cells and most potently promotes angiogenesis. Angiogenesis is critical for the initiation and progression of endometriosis. We established an anti-C16 peptide monoclonal IgM (anti-C16 mAb) by immunizing mice with BSA-conjugated C16 peptide. Anti-C16 mAb inhibited in vitro tube formation of human umbilical endothelial cells (HUVECs) and in vivo angiogenesis in nude mice. We will investigate the inhibitory effect of anti-C16 mAb on angiogenesis in human endometrial fragments or endometriotic lesions in vitro. Less
|
