Clinical & Experimental Metastasis

, Volume 25, Issue 3, pp 233–240

Immunomagnetic enrichment of disseminated tumor cells in bone marrow and blood of breast cancer patients by the Thomsen-Friedenreich-Antigen

Authors

    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
  • Julia Stellwagen
    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
  • Udo Jeschke
    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
  • Uwe Karsten
    • Max Delbrück Center for Molecular Medicine
  • Brigitte Rack
    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
  • Wolfgang Janni
    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
  • Julia Jückstock
    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
  • Augustinos Tulusan
    • Frauenklinik, Klinikum Bayreuth
  • Harald Sommer
    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
  • Klaus Friese
    • First Department of Obstetrics & GynecologyLudwig Maximilians University of Munich
Research Paper

DOI: 10.1007/s10585-007-9137-z

Cite this article as:
Schindlbeck, C., Stellwagen, J., Jeschke, U. et al. Clin Exp Metastasis (2008) 25: 233. doi:10.1007/s10585-007-9137-z

Abstract

Purpose The presence of disseminated tumor cells in the bone marrow (DTC-BM) of breast cancer patients has shown independent prognostic impact. Immunomagnetic enrichment of such cells is an approach to increase the number of detected cells with limited sample volume, especially for circulating tumor cells (CTCs) in blood. The Thomsen-Friedenreich (TF) antigen (CD 176) is a specific oncofetal carbohydrate epitope (Galβ1-3GalNAcα-O) expressed on the surface of various carcinomas. Own studies demonstrated a nearly complete TF expression on DTC-BM, indicating its suitability as marker for immunomagnetic enrichment. Methods BM samples of 65 and peripheral blood samples of 11 breast cancer patients were examined immunocytochemically by staining with the anti-Cytokeratin antibody A45-B/B3 before and after immunomagnetic enrichment. Enrichment was done by incubation with the primary antibody TF 2 (IgM), followed by secondary magnetically labelled rat-anti mouse IgM. Cytospin slides were screened manually by bright-field microscopy. Results 15/65 pts (23%) showed DTC-BM in primary screening with a median of 2/2 mio cells (range 1–10). By enrichment, a median of 23.3 mio cells (0.8–218) could be analysed, increasing positivity to 72% (47/65 pts) with a med. of 4 DTCs (1–105, P < .0001). Blood from 1/11 pts before and 5/11 (45%) after enrichment showed CTCs (med. 2, 1–20), at a med. of 12.4 mio (2.6–38.5) cells analysed. Comparing BM and blood of the same patients after enrichment, 5 were positive in both compartments, 4 showed DTC-BM without presence of CTCs. Conclusion The positive immunomagnetic enrichment technique with TF-antibodies enables to analyse larger sample volumes and increase tumor cell detection rate. This could allow monitoring and characterisation of CTCs as targets for therapies.

Keywords

BloodBone marrowBreast cancerDisseminated tumor cellsEnrichmentThomsen-Friedenreich-Antigen

Abbreviations

APAAP

Alkaline phosphatase anti-alkaline phosphatase

BM

Bone marrow

CK

Cytokeratin

CTC

Circulating tumor cells

DTC

Disseminated tumor cells

EpCam

Epithelial cell adhesion molecule

FCS

Fetal calf serum

HER2

Human epidermal growth factor receptor 2

MACS

Magnetic activated cell sorting

MNC

Mononuclear cells

PB

Peripheral blood

PBS

Phosphate buffered saline

PCR

Polymerase chain reaction

TF

Thomsen-Friedenreich

Copyright information

© Springer Science+Business Media B.V. 2008