Sentinel node lymphocytes: tumour reactive lymphocytes identified intraoperatively for the use in immunotherapy of colon cancer

Abstract

The sentinel node is the first lymph node to receive lymphatic drainage from a tumour and is usually the first site of metastases. Today, the sentinel node is used for tumour staging. Here, we focus on its immunological role and investigate lymphocytic function in sentinel nodes, identified intraoperatively by peritumoural dye injection, from 15 patients with colon cancer. Tumour infiltrating lymphocytes, sentinel and nonsentinel lymph node cells and peripheral blood leukocytes were studied by flow cytometry, proliferation assays and interferon-g secretion after activation with autologous tumour homogenate. Whereas tumour-infiltrating lymphocytes were nonresponsive in the proliferation assays, lymphocytes from sentinel nodes proliferated dose dependently and secreted interferon-g upon stimulation with tumour homogenate. The responses were of varying magnitude and tended to be weaker in metastatic sentinel nodes. Sentinel node lymphocytes represents an enriched source of tumour reactive lymphocytes, and may be useful in future trials of adoptive immunotherapy.

CpG Methylation of the IFNG Gene as a Mechanism to Induce Immunosupression in Tumor-Infiltrating Lymphocytes

“…5C) and not committed to the Th1 phenotype. Interestingly, when we investigate the functionality of TILs from patients with colon cancer they are frequently anergic and unable to produce IFN-␥ upon activation with autologous tumor extract (16). Furthermore we show in this study that the limited IFN-␥ expression in TILs is methylation dependent, since treatment with 5-Azacytidine is able to restore IFN-␥ production in these cells (Fig.…”

“…We have previously shown that tumor-infiltrating lymphocytes are poor producers of IFN-␥ when stimulated with autologous tumor extract or ConA (16). The observation that TILs are hypermethylated at the IFN-␥ (Fig.…”

“…Tumor reactive lymphocytes encounter their Ags in the tumor draining lymph node (16). The limited IFN-␥ production (16) and hypermethylation ( (Fig.…”

“…Tumors from six patients with colon cancer were cut and suspended using a loose fit glass homogenizer to obtain single cell suspensions. During surgery, tumor draining lymph nodes were identified as described previously (16). Tumor-infiltrating CD4 ϩ T lymphocytes and CD4 ϩ cells from tumor draining lymph node were extracted using a CD4 recognizing magnetically labeled Ab and columns according to manufacturer's description (Miltenyi Biotec).…”

Multiplex B Cell Characterization in Blood, Lymph Nodes, and Tumors from Patients with Malignancies

“…Yet, it remains to be demonstrated whether they play this role in humans with solid tumors. We have previously described that a tumor-specific Th1 response is present in sentinel LNs in colon and bladder cancer (42,43), and that sentinel node-acquired CD4 + T cells are promising for use in adoptive immunotherapies (1). In this study, we emulated sentinel node-resident total APCs for ex vivo T cell activation and expansion.…”