Sampling tumor-draining lymph nodes for phenotypic and functional analysis of dendritic cells and T cells.
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.
Vuylsteke
Ronald J C L M
RJ
Department of Surgical Oncology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands.
van Leeuwen
Paul A M
PA
Meijer
Sybren
S
Wijnands
Pepijn G J T B
PG
Statius Muller
Markwin G
MG
Busch
Dirk H
DH
Scheper
Rik J
RJ
de Gruijl
Tanja D
TD
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*
D015789:HLA-A2 Antigen; D018719:Erb-b2 Receptor Tyrosine Kinases