Immunohistochemical detection of an AGE, a ligand for macrophage receptor, in peritoneum of CAPD patients.
BACKGROUND
In patients on long-term continuous ambulatory peritoneal dialysis (CAPD), ultrafiltration (UF) capacity of peritoneal membrane may be impaired due to accumulation of advanced glycation end products (AGEs). This study aimed to elucidate the characteristics of a novel anti-AGE antibody, ODI-GLC19, and to demonstrate AGE accumulation in the peritoneum of CAPD patients using the antibody.


METHODS
A monoclonal anti-AGE antibody (ODI-GLC19) was prepared by immunizing female balb/c mice using D-glucose-modified keyhole limpet hemocyanin. The characteristics of ODI-GLC19 were determined by enzyme-linked immunosorbent assay and receptor binding inhibition assay. Immunohistochemistry using ODI-GLC19 was performed to detect AGE in peritoneal tissues obtained from patients with nonrenal disease, and CAPD patients with normal and low UF.


RESULTS
ODI-GLC19 reacted with glycolaldehyde-modified BSA (GA-BSA) and glucose-modified BSA (GLC-BSA), but not with imidazolone or N epsilon-(carboxymethyl)lysine. GA-BSA and GLC-BSA strongly bound to cultured macrophages. Time-dependent recognition of newly formed GA-BSA by ODI-GLC19 was similar to that by macrophages. The binding of GA-BSA to macrophages was inhibited by ODI-GLC19 in a dose-dependent manner. Immunohistochemical studies revealed that ODI-GLC19-positive AGE was exclusively detected in peritoneal cells including macrophages, and its staining intensity was more prominent in the peritoneum of CAPD patients, especially with low UF, than in patients with nonrenal disease.


CONCLUSIONS
A novel monoclonal anti-AGE antibody, ODI-GLC19, recognizes a ligand for an AGE receptor on macrophages. Incorporation of AGE into peritoneal cells including macrophages may be involved in progressive peritoneal dysfunction in CAPD patients.
Nakamura
Sakurako
S
Department of Clinical Preventive Medicine, Nagoya University Hospital, Nagoya, Japan.
Tobita
Kazuki
K
Tachikawa
Tetsuya
T
Akamatsu
Suguru
S
Ohno
Yoshifumi
Y
Kan
Ayumi
A
Katsuragawa
Misaki
M
Yasumura
Koichi
K
Miyazaki
Shigeru
S
Sakai
Shinji
S
Morita
Takashi
T
Hirashawa
Yoshihei
Y
Niwa
Toshimitsu
T
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D017127:Glycation End Products, Advanced / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D006433:Hemocyanins / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D007457:Iodine Radioisotopes; D007676:Kidney Failure, Chronic / Q000378:metabolism* / Q000628:therapy; D008024:Ligands; D017737:Macrophages, Peritoneal / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D010531:Peritoneal Dialysis, Continuous Ambulatory; D010537:Peritoneum / Q000166:cytology / Q000378:metabolism; D011869:Radioligand Assay; D011956:Receptors, Cell Surface / Q000378:metabolism
D000911:Antibodies, Monoclonal; D017127:Glycation End Products, Advanced; D007457:Iodine Radioisotopes; D008024:Ligands; D011956:Receptors, Cell Surface; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin