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

Ronald J.C.L.M. Vuylsteke, Paul A.M. Van Leeuwen*, Sybren Meijer, Pepijn G.J.T.B. Wijnands, Markwin G.Statius Muller, Dirk H. Busch, Rik J. Scheper, Tanja D. De Gruijl

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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. Tumordraining 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)-γ 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.

Original languageEnglish
Article number64152
Pages (from-to)19-26
Number of pages8
JournalAmerican Journal of Pathology
Volume161
Issue number1
DOIs
Publication statusPublished - 1 Jan 2002

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