title: Establishment of optimized ELISA system specific for HLA-G in body fluids.
abstract: Recently, human leukocyte antigen-G (HLA-G) has been a focus in the field of reproductive immunology, tumor progression and transplantation, because of its inhibitory function as ligand to the inhibitory receptors leukocyte immunoglobulin-like receptors (LILR) B1 and LILRB2. The HLA-G is expressed in distinct mRNA isoforms, one of which encodes a soluble HLA-G (sHLA-G) protein, detectable by sandwich ELISA. Therefore, sHLA-G ELISAs have been used as a noninvasive diagnosis system. While a number of sHLA-G-specific ELISAs have been described, our prior studies showed that data obtained by the conventional ELISA system detecting sHLA-G in body fluids was not consistent with the data obtained from immunoprecipitation (IP)/immunoblotting (IB). Therefore, we established an optimized ELISA system described in this report, which yields results consistent with IP/IB analysis. Using this system, we determined sHLA-G protein in amniotic fluids, and found that sHLA-G levels at preterm (∼36 weeks) were clearly higher than those at term (37-41 weeks). These data and supporting experiments showed that the ELISA system we established can be an useful tools for the detection of sHLA-G protein in body fluids than the conventional ELISA system.
authors: Department of Immunology, Nara Medical University, Kashihara, Japan.
authors: N
authors: N
authors: Ouji-Sageshima
authors: The Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
authors: D E
authors: DE
authors: Geraghty
authors: Department of Legal Medicine, Nara Medical University, Kashihara, Japan.
authors: A
authors: A
authors: Ishitani
authors: Department of Legal Medicine, Nara Medical University, Kashihara, Japan.
authors: K
authors: K
authors: Hatake
authors: Department of Immunology, Nara Medical University, Kashihara, Japan.
authors: T
authors: T
authors: Ito
mesh_terms: D000653:Amniotic Fluid / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001402:B-Lymphocytes; D015153:Blotting, Western; D002461:Cell Line, Transformed; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards*; D005260:Female; D005333:Fetus; D059951:HLA-G Antigens / Q000097:blood / Q000302:isolation & purification*; D006801:Humans; D047468:Immunoprecipitation; D007231:Infant, Newborn; D007234:Infant, Premature; D011247:Pregnancy; D020033:Protein Isoforms / Q000097:blood / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification; D012995:Solubility
chemical_list: D000911:Antibodies, Monoclonal; D059951:HLA-G Antigens; D020033:Protein Isoforms; D011994:Recombinant Proteins
keywords: ELISA; HLA-G; follicular fluids; placental lavage fluids; plasma