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(American Journal of Pathology. 2002;161:19-26.)
© 2002 American Society for Investigative Pathology

Technical Advances

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{dagger}, Markwin G. Statius Muller*, Dirk H. Busch{ddagger}, Rik J. Scheper{dagger} and Tanja D. de Gruijl§

From the Departments of Surgical Oncology,*Pathology,{dagger}and Medical Oncology,§Vrije Universiteit Medical Center, Amsterdam, The Netherlands; and the Institute of Medical Microbiology, Immunology, and Hygiene,{ddagger}Technical University Munich, Munich, Germany

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.




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