A sandwich ELISA for detection of soluble GPI-80, a glycosylphosphatidyl-inositol (GPI)-anchored protein on human leukocytes involved in regulation of neutrophil adherence and migration--its release from activated neutrophils and presence in synovial fluid of rheumatoid arthritis patients.
GPI-80, mainly distributed on human neutrophils and monocytes, is a novel glycosylphosphatidylinositol (GPI)-anchored protein regulating neutrophil adherence and migration. We prepared a polyclonal antibody specific to GPI-80 by immunizing rabbits with the purified GPI-80 and established a sandwich ELISA for detecting soluble GPI-80. We found that soluble GPI-80 was released from fMLP activated neutrophils and was present at high concentrations in synovial fluids but not sera of rheumatoid arthritis patients, suggesting that GPI-80 may play a role in inflammatory diseases. The detection of soluble GPI-80 may help us to understand its physiological and pathological functions.
Huang
J
J
Department of Immunology and Parasitology, Yamagata University School of Medicine, Yamagata, Japan.
Takeda
Y
Y
Watanabe
T
T
Sendo
F
F
D000581:Amidohydrolases; D000818:Animals; D000911:Antibodies, Monoclonal; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000378:metabolism*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D002465:Cell Movement; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D058851:GPI-Linked Proteins; D006801:Humans; D006867:Hydrolases; D007106:Immune Sera; D008213:Lymphocyte Activation; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D013582:Synovial Fluid / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D058851:GPI-Linked Proteins; D007106:Immune Sera; D006867:Hydrolases; D000581:Amidohydrolases; C121771:VNN2 protein, human