Sampling tumor-draining lymph nodes for phenotypic and functional analysis of dendritic cells and T cells

Am J Pathol. 2002 Jul;161(1):19-26. doi: 10.1016/S0002-9440(10)64152-1.


Immune responses against tumor antigens will initially occur in the first tumor-draining lymph node, the sentinel node (SN). Because of extensive diagnostic procedures, obtaining a piece of SN to isolate viable immune cells for functional analyses is often impossible. For this reason an alternative method to obtain viable cells from a lymph node (LN) was investigated, ie, scraping LNs with a surgical blade, and compared with dissociation of total LNs. Tumor-draining lymph nodes were retrieved from five oncological patients. The collected dendritic cells and T cells were phenotypically and functionally characterized by flow cytometry and antigen-specific interferon (IFN)-gamma release in an ELISPOT assay. Results were compared between the two isolation methods. Viabilities and phenotypic characteristics of the collected cells were entirely comparable for both methods. T-cell functionality was also comparable between both methods, with equal T-cell expansion factors and similar frequencies of cytotoxic T cells specifically recognizing the M1 matrix protein of Influenza haemophilus or the tumor antigen Her-2/neu. In conclusion, scraping LNs to obtain cells for analysis of immune functions in LNs is feasible and presents a good alternative to dissociation of LNs. Scraping may even be applied to small LNs that a pathologist will submit entirely for histological examination and may thus prove useful in the monitoring of immune responses in SNs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / metabolism
  • CD4-CD8 Ratio
  • CD4-Positive T-Lymphocytes / physiology*
  • CD8-Positive T-Lymphocytes / physiology*
  • Cell Survival / physiology
  • Dendritic Cells / physiology*
  • Female
  • HLA-A2 Antigen / metabolism
  • Humans
  • Lymph Nodes / pathology
  • Lymph Nodes / physiopathology*
  • Neoplasms / pathology
  • Neoplasms / physiopathology*
  • Phenotype
  • Receptor, ErbB-2 / metabolism
  • Specimen Handling / methods*


  • HLA-A2 Antigen
  • Receptor, ErbB-2