EPISPOT Assay: Detection of Viable DTCs/CTCs in Solid Tumor Patients

  • Catherine Alix-Panabières
Part of the Recent Results in Cancer Research book series (RECENTCANCER, volume 195)


The enumeration and characterization of circulating tumor cells (CTCs) in the peripheral blood and disseminated tumor cells (DTCs) in bone marrow may provide important prognostic information and might help to monitor efficacy of therapy. Since current assays cannot distinguish between apoptotic and viable DTCs/CTCs, it is now possible to apply a novel ELISPOT assay (designated ‘EPISPOT’) that detects proteins secreted/released/shed from single epithelial cancer cells. Cells are cultured for a short time on a membrane coated with antibodies that capture the secreted/released/shed proteins which are subsequently detected by secondary antibodies labeled with fluorochromes. In breast cancer, we measured the release of cytokeratin-19 (CK19) and mucin-1 (MUC1) and demonstrated that many patients harbored viable DTCs, even in patients with apparently localized tumors (stage M0: 54%). Preliminary clinical data showed that patients with DTC-releasing CK19 have an unfavorable outcome. We also studied CTCs or CK19-secreting cells in the peripheral blood of M1 breast cancer patients and showed that patients with CK19-SC had a worse clinical outcome. In prostate cancer, we used prostate-specific antigen (PSA) secretion as marker and found that a significant fraction of CTCs secreted fibroblast growth factor-2 (FGF2), a known stem cell growth factor. In conclusion, the EPISPOT assay offers a new opportunity to detect and characterize viable DTCs/CTCs in cancer patients and it can be extended to a multi-parameter analysis revealing a CTC/DTC protein fingerprint.


Overt Metastasis Large Blood Volume Specific Protein Marker Stem Cell Growth Factor Hepatocellular Cancer Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Alix-Panabieres C, Muller V, Pantel K (2007) Current status in human breast cancer micrometastasis. Curr Opin Oncol 19:558–563PubMedCrossRefGoogle Scholar
  2. 2.
    Alix-Panabieres C, Riethdorf S, Pantel K (2008) Circulating tumor cells and bone marrow micrometastasis. Clin Cancer Res 14:5013–5021PubMedCrossRefGoogle Scholar
  3. 3.
    Alix-Panabieres C et al (2005) Characterization and enumeration of cells secreting tumor markers in the peripheral blood of breast cancer patients. J Immunol Methods 299:177–188PubMedCrossRefGoogle Scholar
  4. 4.
    Alix-Panabieres C et al (2005) Detection of circulating prostate-specific antigen-secreting cells in prostate cancer patients. Clin Chem 51:1538–1541PubMedCrossRefGoogle Scholar
  5. 5.
    Pantel K, Alix-Panabieres C (2007) The clinical significance of circulating tumor cells. Nat Clin Pract Oncol 4:62–63PubMedCrossRefGoogle Scholar
  6. 6.
    Pantel K, Alix-Panabieres C, Riethdorf S (2009) Cancer micrometastases. Nat Rev Clin Oncol 6:339–351PubMedCrossRefGoogle Scholar
  7. 7.
    Pantel K (2010) Circulating tumour cells in cancer patients: challenges and perspectives. Trends Mol Med 16:398–406PubMedCrossRefGoogle Scholar
  8. 8.
    Czerkinski C, Nilsson LA, Nygren H, Ouchterlony O, Tarkowski A (1983) A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells. J Immunol Methods 65:109–121CrossRefGoogle Scholar
  9. 9.
    Solakoglu O et al (2002) Heterogeneous proliferative potential of occult metastatic cells in bone marrow of patients with solid epithelial tumors. Proc Natl Acad Sci U S A 99:2246–2251PubMedCrossRefGoogle Scholar
  10. 10.
    Alix-Panabieres C et al (2007) Detection and characterization of putative metastatic precursor cells in cancer patients. Clin Chem 53:537–539PubMedCrossRefGoogle Scholar
  11. 11.
    Ho SB et al (1993) Heterogeneity of mucin gene expression in normal and neoplastic tissues. Cancer Res 53:641–651PubMedGoogle Scholar
  12. 12.
    Mommers EC et al (1999) Aberrant expression of MUC1 mucin in ductal hyperplasia and ductal carcinoma In situ of the breast. Int J Cancer 84:466–469PubMedCrossRefGoogle Scholar
  13. 13.
    Parry S et al (2001) Identification of MUC1 proteolytic cleavage sites in vivo. Biochem Biophys Res Commun 283:715–720PubMedCrossRefGoogle Scholar
  14. 14.
    Chu PG, Weiss LM (2002) Keratin expression in human tissues and neoplasms. Histopathology 40:403–439PubMedCrossRefGoogle Scholar
  15. 15.
    Zhou X, Liao J, Hu L, Feng L, Omary MB (1999) Characterization of the major physiologic phosphorylation site of human keratin 19 and its role in filament organization. J Biol Chem 274:12861–12866PubMedCrossRefGoogle Scholar
  16. 16.
    Zach O, Lutz D (2006) Tumor cell detection in peripheral blood and bone marrow. Curr Opin Oncol 18:48–56PubMedCrossRefGoogle Scholar
  17. 17.
    Gudjonsson T et al (2002) Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. Genes Dev 16:693–706PubMedCrossRefGoogle Scholar
  18. 18.
    Alix-Panabieres C et al (2009) Full-length cytokeratin-19 is released by human tumor cells: a potential role in metastatic progression of breast cancer. Breast Cancer Res 11:R39PubMedCrossRefGoogle Scholar
  19. 19.
    Doyen J et al (2011) Circulating tumor cells in prostate cancer: A potential surrogate marker of survival. Crit Rev Oncol HematolGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  1. 1.Laboratory of Rare Human Circulating CellsSaint-Eloi Hospital, University Medical Centre, Institute of Research in Biotherapy, University Montpellier 1MontpellierFrance
  2. 2.Laboratory of Cell and Hormonal BiologyArnaud de Villeneuve Hospital, University Medical Centre, University Montpellier 1MontpellierFrance
  3. 3.Biostatistics and Public HealthUniversity Institute of Clinical Research UM1—EA2415—EpidemiologyMontpellierFrance

Personalised recommendations