title: Comparative cellular catabolism and retention of astatine-, bismuth-, and lead-radiolabeled internalizing monoclonal antibody.
abstract: Monoclonal antibodies (mAbs) labeled with alpha-emitting radionuclides such as (211)At, (212)Bi, (213)Bi, and (212)Pb (which decays by beta-emission to its alpha-emitting daughter, (212)Bi) are being evaluated for their potential applications for cancer therapy. The fate of these radionuclides after cells are targeted with mAbs is important in terms of dosimetry and tumor detection.
  
  
  METHODS
  In this study, we attached various radionuclides that result in alpha-emissions to T101, a rapidly internalizing anti-CD5 mAb. We then evaluated the catabolism and cellular retention and compared them with those of (125)I- and (111)In-labeled T101. T101 was labeled with (211)At, (125)I, (205,6)Bi, (111)In, and (203)Pb. CD5 antigen-positive cells, peripheral blood mononuclear cells (PBMNC), and MOLT-4 leukemia cells were used. The labeled T101 was incubated with the cells for 1 h at 4 degrees C for surface labeling. Unbound activity was removed and 1 mL medium added. The cells were then incubated at 37 degrees C for 0, 1, 2, 4, 8, and 24 h. The activity on the cell surface that internalized and the activity on the cell surface remaining in the supernatant were determined. The protein in the supernatant was further precipitated by methanol for determining protein-bound and non-protein-bound radioactivity. Sites of internal cellular localization of radioactivity were determined by Percoll gradient centrifugation.
  
  
  RESULTS
  All radiolabeled antibodies bound to the cells were internalized rapidly. After internalization, (205,6)Bi, (203)Pb, and (111)In radiolabels were retained in the cell, with little decrease of cell-associated radioactivity. However, (211)At and (125)I were released from cells rapidly ((211)At < (125)I) and most of the radioactivity in the supernatant was in a non-protein-bound form. Intracellular distribution of radioactivity revealed a transit of the radiolabel from the cell surface to the lysosome. The catabolism patterns of MOLT-4 cells and PBMNC were similar.
  
  
  CONCLUSIONS
  (211)At catabolism and release from cells were somewhat similar to that of (125)I, whereas (205,6)Bi and (203)Pb showed prolonged cell retention similar to that of (111)In. These catabolism differences may be important in the selection of alpha-radionuclides for radioimmunotherapy.
authors.affiliation: Department of Nuclear Medicine, Warren G. Magnuson Clinical Center, National Institutes of Health, 10 Center Dr., Bethesda, MD 20892-1180, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Yao
authors.forename: K
authors.initials: K
authors.lastname: Garmestani
authors.forename: K J
authors.initials: KJ
authors.lastname: Wong
authors.forename: L S
authors.initials: LS
authors.lastname: Park
authors.forename: E
authors.initials: E
authors.lastname: Dadachova
authors.forename: A
authors.initials: A
authors.lastname: Yordanov
authors.forename: T A
authors.initials: TA
authors.lastname: Waldmann
authors.forename: W C
authors.initials: WC
authors.lastname: Eckelman
authors.forename: C H
authors.initials: CH
authors.lastname: Paik
authors.forename: J A
authors.initials: JA
authors.lastname: Carrasquillo
mesh_terms: D000512:Alpha Particles; D000911:Antibodies, Monoclonal / Q000493:pharmacokinetics*; D001246:Astatine / Q000493:pharmacokinetics*; D001729:Bismuth / Q000493:pharmacokinetics*; D018956:CD5 Antigens / Q000276:immunology; D006801:Humans; D018796:Immunoconjugates / Q000493:pharmacokinetics*; D007205:Indium Radioisotopes / Q000493:pharmacokinetics; D007457:Iodine Radioisotopes / Q000493:pharmacokinetics; D007856:Lead Radioisotopes / Q000493:pharmacokinetics*; D011868:Radioisotopes / Q000493:pharmacokinetics*; D014407:Tumor Cells, Cultured / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D018956:CD5 Antigens; D018796:Immunoconjugates; D007205:Indium Radioisotopes; D007457:Iodine Radioisotopes; D007856:Lead Radioisotopes; D011868:Radioisotopes; D001729:Bismuth; D001246:Astatine