This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Méhes, G. Articles by Ambros, P. F. Search for Related Content PubMed PubMed Citation Articles by Méhes, G. Articles by Ambros, P. F.

Combined Automatic Immunological and Molecular Cytogenetic Analysis Allows Exact Identification and Quantification of Tumor Cells in the Bone Marrow

Children’s Cancer Research Institute, St. Anna Kinderspital, A-1090 Vienna, Austria

Purpose: To improve the detection of disseminated tumor cells in bone marrow (BM) and peripheral blood samples of solid tumor patients, a novel computer-assisted scanning system for automatic search, image analysis, and repositioning of these cells was developed. This system allows precise identification and quantification of tumor cells by sequential immunological and molecular cytogenetic analysis. In this study, we attempt to demonstrate the practical use of this approach by analyzing BM samples from neuroblastoma patients.

Experimental Design: The disialo-ganglioside (GD2) molecule was used as the immunological target. The GD2 molecule was described as being specific for neuroblastoma cells, although false positive reactions had been suspected. To verify or disprove the neoplastic nature of the immunologically positive cells, sequential fluorescence in situ hybridization was performed on these cells to search for those genetic aberrations found in the corresponding primary tumors. A total of 115 samples from 40 newly diagnosed patients were evaluated for the presence of GD2⁺ cells in the BM.

Results: GD2 positivity was detected in 95.2% of stage 4 patients, in 100% of stage 4s patients, and in 38.5% of patients with localized/regional disease. In stage 4 and 4s BM samples, the GD2⁺ cells were unequivocally identified as tumor cells based on the molecular cytogenetic aberrations found by fluorescence in situ hybridization. However, in BM samples from patients with localized/regional disease, all GD2⁺ cells were concluded to represent false positivity due to the absence of genetic aberrations.

Conclusions: Automatic search and sequential molecular cytogenetic analysis of the immunologically positive cells provide precise information on both the number and cytogenetic profile of disseminated tumor cells.

This article has been cited by other articles:

				J. Stutterheim, A. Gerritsen, L. Zappeij-Kannegieter, B. Yalcin, R. Dee, M. M. van Noesel, F. Berthold, R. Versteeg, H. N. Caron, C. E. van der Schoot, et al. Detecting Minimal Residual Disease in Neuroblastoma: The Superiority of a Panel of Real-Time Quantitative PCR Markers Clin. Chem., July 1, 2009; 55(7): 1316 - 1326. [Abstract] [Full Text] [PDF]

				J. Stutterheim, A. Gerritsen, L. Zappeij-Kannegieter, I. Kleijn, R. Dee, L. Hooft, M. M. van Noesel, M. Bierings, F. Berthold, R. Versteeg, et al. PHOX2B Is a Novel and Specific Marker for Minimal Residual Disease Testing in Neuroblastoma J. Clin. Oncol., November 20, 2008; 26(33): 5443 - 5449. [Abstract] [Full Text] [PDF]

				C. Alix-Panabieres, S. Riethdorf, and K. Pantel Circulating Tumor Cells and Bone Marrow Micrometastasis Clin. Cancer Res., August 15, 2008; 14(16): 5013 - 5021. [Abstract] [Full Text] [PDF]

				K. Swerts, P. F. Ambros, C. Brouzes, J. M. F. Navarro, N. Gross, D. Rampling, R. Schumacher-Kuckelkorn, A. R. Sementa, R. Ladenstein, and K. Beiske Standardization of the Immunocytochemical Detection of Neuroblastoma Cells in Bone Marrow J. Histochem. Cytochem., December 1, 2005; 53(12): 1433 - 1440. [Abstract] [Full Text] [PDF]

				S. Meng, D. Tripathy, E. P. Frenkel, S. Shete, E. Z. Naftalis, J. F. Huth, P. D. Beitsch, M. Leitch, S. Hoover, D. Euhus, et al. Circulating Tumor Cells in Patients with Breast Cancer Dormancy Clin. Cancer Res., December 15, 2004; 10(24): 8152 - 8162. [Abstract] [Full Text] [PDF]

				S.-K. Kraeft, A. Ladanyi, K. Galiger, A. Herlitz, A. C. Sher, D. E. Bergsrud, G. Even, S. Brunelle, L. Harris, R. Salgia, et al. Reliable and Sensitive Identification of Occult Tumor Cells Using the Improved Rare Event Imaging System Clin. Cancer Res., May 1, 2004; 10(9): 3020 - 3028. [Abstract] [Full Text] [PDF]

				M M Reid Minimal metastatic disease J. Clin. Pathol., January 1, 2004; 57(1): 21 - 21. [Full Text] [PDF]

				K. Pantel, V. Muller, M. Auer, N. Nusser, N. Harbeck, and S. Braun Detection and Clinical Implications of Early Systemic Tumor Cell Dissemination in Breast Cancer Clin. Cancer Res., December 15, 2003; 9(17): 6326 - 6334. [Abstract] [Full Text] [PDF]

				G. Mehes, A. Luegmayr, R. Kornmuller, I. M. Ambros, R. Ladenstein, H. Gadner, and P. F. Ambros Detection of Disseminated Tumor Cells in Neuroblastoma: 3 Log Improvement in Sensitivity by Automatic Immunofluorescence plus FISH (AIPF) Analysis Compared with Classical Bone Marrow Cytology Am. J. Pathol., August 1, 2003; 163(2): 393 - 399. [Abstract] [Full Text] [PDF]

				J. I. Martin-Subero, I. Chudoba, L. Harder, S. Gesk, W. Grote, F. J. Novo, M. J. Calasanz, and R. Siebert Multicolor-FICTION: Expanding the Possibilities of Combined Morphologic, Immunophenotypic, and Genetic Single Cell Analyses Am. J. Pathol., August 1, 2002; 161(2): 413 - 420. [Abstract] [Full Text] [PDF]