title: Sampling tumor-draining lymph nodes for phenotypic and functional analysis of dendritic cells and T cells.
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. 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.
authors.affiliation: Department of Surgical Oncology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands.
authors.forename: Ronald J C L M
authors.initials: RJ
authors.lastname: Vuylsteke
authors.forename: Paul A M
authors.initials: PA
authors.lastname: van Leeuwen
authors.forename: Sybren
authors.initials: S
authors.lastname: Meijer
authors.forename: Pepijn G J T B
authors.initials: PG
authors.lastname: Wijnands
authors.forename: Markwin G
authors.initials: MG
authors.lastname: Statius Muller
authors.forename: Dirk H
authors.initials: DH
authors.lastname: Busch
authors.forename: Rik J
authors.initials: RJ
authors.lastname: Scheper
authors.forename: Tanja D
authors.initials: TD
authors.lastname: de Gruijl
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism; D016516:CD4-CD8 Ratio; D015496:CD4-Positive T-Lymphocytes / Q000502:physiology*; D018414:CD8-Positive T-Lymphocytes / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D003713:Dendritic Cells / Q000502:physiology*; D005260:Female; D015789:HLA-A2 Antigen / Q000378:metabolism; D006801:Humans; D008198:Lymph Nodes / Q000473:pathology / Q000503:physiopathology*; D009369:Neoplasms / Q000473:pathology / Q000503:physiopathology*; D010641:Phenotype; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D013048:Specimen Handling / Q000379:methods*
chemical_list: D015789:HLA-A2 Antigen; D018719:Erb-b2 Receptor Tyrosine Kinases