Generation of tumor-specific cytotoxic T-lymphocytes from the peripheral blood of colorectal cancer patients for adoptive T-cell transfer

Abstract

This study designs a strategy for an adoptive cellular therapy (ACT) protocol based on the ex-vivo selection of autologous peripheral blood-derived CD8-enriched T-cells, stimulated with dendritic cells (DCs) that had been pulsed with apoptotic tumor cells to generate cytotoxic T lymphocytes (CTLs) with anti-tumor activity. Seventy-eight colorectal cancer (CRC) patients were enrolled in this study. Tumor tissues and peripheral blood (PB) were obtained at surgery. Tissues were mechanically dissociated and cultured to obtain a primary tumor cell line from each patient. DCs were derived from peripheral blood mononuclear cells (PBMCs) using magnetic positive selection of CD14+ monocytes. Anti-tumor CTLs were elicited in co-/micro-cultures using DCs as antigen-presenting cells, autologous apoptotic tumor cells as a source of antigens, and CD8+ T lymphocytes as effectors. Interferon-γ (IFN-γ) secretion was assessed by ELISpot assays to evaluate the activation of the CTLs against the autologous tumor cells. Primary tumor cell lines were obtained from 20 of 78 patients (25.6%). DCs were generated from 26 patients, and of them, corresponding tumor cell lines were derived from six patients. ELISpot results showed that significant IFN-γ secretion was detected after different numbers of stimulations for two patients, whereas weak secretion was observed for three patients. Despite difficulties due to contamination of several primary tumor cell lines with gut intestinal flora, the results suggest that the generation of tumor-specific CTLs is feasible from patients with CRC, and could be useful for supporting an ACT approach in CRC.

TCR sequencing analysis of cancer tissues and tumor draining lymph nodes in colorectal cancer patients

“…38 The biggest challenging part of this type of therapy is an identification of antigen (neoantigen and/or shared antigen)-specific T cells. TILs or lymphocytes from PBMCs have been used to induce and identify antigen-specific T cells, 36,[39][40][41] and we reported a method to identify antigen-specific TCRs using PBMCs in as short as two weeks, 36 Since our results suggest that TDLNs with metastatic cancer cells seem to be enriched with tumor-reactive T cells, TDLNs with cancer cells may also be a good source of T lymphocytes to induce antigen-specific T cells in our protocol. In this study, we found that metastasis-positive TDLNs tended to reveal similar TCR repertoire patterns each other (Figure 4, Supplementary Figure 1).…”

Delineation of an immunosuppressive gradient in hepatocellular carcinoma using high-dimensional proteomic and transcriptomic analyses

“…5C) with irradiated autologous tumor samples (as a source of tumor antigens) and feeder cells, as previously described (22). After 48-h exposure to the irradiated tumor sample, the ratio of PD-1 versus T-bet expression on T EM cells from the PBMCs was enhanced compared with day 0, and this ratio was maintained until day 6 (Fig.…”

Disappearance of bone metastases in chemotherapy‑resistant gastric cancer treated with antigen peptide‑pulsed dendritic cell‑activated cytotoxic T lymphocyte immunotherapy: A case report

“…As an immune-based approach, adoptive therapy has become an increasingly attractive modality for cancer therapy due to collectively demonstrating a decreased risk of cross-resistance compared with conventional therapies, high specificity and long-term immune protection ( 15 ). Recent success of adoptive T cell therapy using ex vivo expanded autologous tumor-reactive T cells has increased optimism that this modality may form a specific therapy for patients with advanced-stage disease, including those who are refractory to standard therapies ( 16 , 17 ). Previously, autologous immune cells intravenously administered to patients with GC have resulted in improved survival rates compared with controls, as reported by Zhang et al ( 18 ).…”