In vivo near-infrared fluorescence imaging of carcinoembryonic antigen-expressing tumor cells in mice

Radiology. 2008 Jun;247(3):779-87. doi: 10.1148/radiol.2472070123. Epub 2008 Apr 15.

Abstract

Purpose: To prospectively depict carcinoembryonic antigen (CEA)-expressing tumors in mice with a high-affinity probe consisting of a near-infrared (NIR) fluorochrome and the clinically used anti-CEA antibody fragment arcitumomab.

Materials and methods: This study was approved by the regional animal committee. By coupling a NIR fluorescent (NIRF) cyanine dye (DY-676) to a specific antibody fragment directed against CEA (arcitumomab) and a nonspecific IgG Fab fragment, a bio-optical high-affinity fluorescent probe (anti-CEA-DY-676) and a low-affinity fluorescent probe (FabIgG-DY-676) were designed. The dye-to-protein ratios were determined, and both probes were tested for NIRF imaging in vitro on CEA-expressing LS-174T human colonic adenocarcinoma cells and CEA-nonexpressing A-375 human melanoma cells by using a bio-optical NIR small-animal imager. In vivo data of xenografted LS-174T and A-375 tumors in mice (n = 10) were recorded and statistically analyzed (Student t test).

Results: The dye-to-protein ratios were determined as 3.0-3.5 for both probes. In vitro experiments revealed the specific binding of the anti-CEA-DY-676 probe on CEA-expressing cells as compared with CEA-nonexpressing cells; the FabIgG-DY-676 probe showed a markedly lower binding affinity to cells. In vivo LS-174T tumors xenografted in all mice could be significantly distinguished from A-375 tumors with application of the anti-CEA-DY-676 but not with that of the FabIgG-DY-676 at different times (2-24 hours, P < .005) after intravenous injection of the probes. Semiquantitative analysis revealed maximal fluorescence signals of anti-CEA-DY-676 to CEA-expressing tumors about 8 hours after injection.

Conclusion: Findings of this study indicate the potential use of the high-affinity probe anti-CEA-DY-676 for specific NIRF imaging in in vivo tumor diagnosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / metabolism*
  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Carbocyanines / chemistry*
  • Carcinoembryonic Antigen / metabolism*
  • Colonic Neoplasms / metabolism*
  • Female
  • Immunoglobulin Fab Fragments / chemistry*
  • Mice
  • Mice, Inbred BALB C
  • Polymerase Chain Reaction
  • Prospective Studies
  • Spectrometry, Fluorescence
  • Spectrophotometry, Infrared
  • Whole Body Imaging

Substances

  • Antibodies, Monoclonal
  • Carbocyanines
  • Carcinoembryonic Antigen
  • Immunoglobulin Fab Fragments