Distribution of anticancer antibiotic daunomycin in the rat heart and kidney revealed by immunocytochemistry using monoclonal antibodies.
Two monoclonal antibodies (ADM-1-11 and 79-31 mAbs) were raised against daunomycin (DM) conjugated to bovine serum albumin via the cross-linker N-(gamma-maleimidobutyryloxy)succinimide. The monoclonal antibodies (mAbs) specifically detected DM as well as its analogs doxorubicin and epirubicin, but did not react with other anticancer antibiotics, including pepleomycin, mitomycin C, and actinomycin D. The mAbs reacted strongly with glutaraldehyde-conjugated DM in an enzyme linked immunosorbent assay (ELISA) used as a model system for immunocytochemistry as well as in appropriately pretreated sections of tissues from animals injected with DM. No staining occurred in tissues from uninjected animals. In order to perform DM ICC a number of tissue treatment conditions critical to the detection of low molecular weight substances were employed. Uptake of DM was studied in rats after a single i.v. or i.p. administration of the drug. In the heart, accumulation of DM occurred in nuclei and in the cytoplasm. In the kidney, DM immunoreactivity accumulated in all segments of the nephron except for the proximal tubules. Since the proximal tubules are known to be where a variety of transport systems including P-glycoprotein (Pgp) and organic anion-transporting polypeptides (OATPs) in drug interactions occur, the absence of DM accumulation in these segments may reflect a transport phenomenon depending upon such transporters. The availability of methods to study sites of accumulation of DM offers possibilities for understanding toxic side effects of this drug on the heart and kidney. Moreover, the immunocytochemical methodology developed may prove useful for the localization of other low molecular weight drugs that can be fixed in situ by glutaraldehyde.
Fujiwara
Kunio
K
Department of Applied Life Science, Faculty of Biotechnology and Life Science, Sojo University, Ikeda, 4-22-1, Kumamoto 860-0082, Japan. fujiwara@life.sojo-u.ac.jp
Shin
Masashi
M
Hougaard
David M
DM
Larsson
Lars-Inge
LI
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000493:pharmacokinetics; D000911:Antibodies, Monoclonal; D003630:Daunorubicin / Q000493:pharmacokinetics*; D007150:Immunohistochemistry; D007668:Kidney / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D014018:Tissue Distribution
D000903:Antibiotics, Antineoplastic; D000911:Antibodies, Monoclonal; D003630:Daunorubicin