title: Immunocytochemistry for vancomycin using a monoclonal antibody that reveals accumulation of the drug in rat kidney and liver.
abstract: We prepared monoclonal antibodies against N-(γ-maleimidobutyryloxy)succinimide-conjugated vancomycin (VM). The monoclonal antibody was specific for conjugated or free VM. The monoclonal antibody enabled us to develop an immunocytochemical method for detecting the uptake of VM in the rat kidney and liver. Three hours after a single intravenous (i.v.) injection of VM at the therapeutic dose, the immunocytochemistry revealed that VM accumulated in large amounts in both the S1 and S2 segments and in much smaller amounts in the S3 segment of the proximal tubules as well as in the distal tubules and collecting ducts. The drug was detected in the cytoplasm, cytoplasmic irregular granules, nuclei, and microvilli of the proximal tubule cells. The distal tubules and collecting ducts contained scattered swollen cells in which both the nuclei and cytoplasm were heavily immunostained. Twenty-four hours after injection, most of the swollen cells returned back to normal size and had somewhat decreased immunostaining. Also, significant amounts of VM remained accumulated for as long as 8 days postadministration. In the liver, similar drug accumulation was observed in the Kupffer cells and the endothelial cells of the hepatic sinusoids but not in the hepatocytes, suggesting that vancomycin cannot be eliminated via the liver. Immunoelectron microscopic studies demonstrated that in the collecting ducts, uptake of VM occurred exclusively in the lysosomes and cytoplasm of the principal cells and scarcely in the intercalated cells. Furthermore, double fluorescence staining using rats simultaneously administered with VM and gentamicin strongly suggests that both drugs colocalized in lysosomes in the proximal tubule cells of kidneys.
authors.affiliation: Department of Applied Life Science, Faculty of Biotechnology and Life Science, Sojo University, Ikeda, Kumamoto, Japan. fujiwara@life.sojo-u.ac.jp
authors.forename: Kunio
authors.initials: K
authors.lastname: Fujiwara
authors.forename: Yohei
authors.initials: Y
authors.lastname: Yoshizaki
authors.forename: Masashi
authors.initials: M
authors.lastname: Shin
authors.forename: Tsubasa
authors.initials: T
authors.lastname: Miyazaki
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Saita
authors.forename: Shuichi
authors.initials: S
authors.lastname: Nagata
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000493:pharmacokinetics*; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007150:Immunohistochemistry; D007275:Injections, Intravenous; D007685:Kidney Tubules, Collecting / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D007728:Kupffer Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008247:Lysosomes / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D051381:Rats; D017208:Rats, Wistar; D013388:Succinimides / Q000737:chemistry*; D014640:Vancomycin / Q000737:chemistry / Q000493:pharmacokinetics*
chemical_list: D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D013388:Succinimides; D014640:Vancomycin; C032138:N-(gamma-maleimidobutyryloxy)succinimide