Cytotoxic effector cells with antitumor activity can be amplified ex vivo from biopsies or blood of patients with renal cell carcinoma for cell therapy use

Abstract

Adoptive immunotherapy with antitumor effector cells is an attractive therapeutic approach in metastatic renal cell carcinoma (RCC). The aim of the work was to enhance in vitro activation of lymphocytes with optimal cytotoxic activity against tumor cells. We evaluated a procedure based on the use of dendritic cells (DCs) loaded with irradiated tumor cells (DC-Tu) to stimulate lymphocytes. Experimental conditions were established with cells from healthy donors and melanoma cell lines. Procedures were then applied to cells from RCC patients. A total of 30 tumor biopsies, 14 proximal lymph nodes, and 17 peripheral blood samples from 30 patients were used. When lymphocytes were stimulated in vitro with DC-Tu, they responded to tumor cells with an increased cytolytic activity for all the assays with donor cells (n=18). For RCC patients, DC-Tu stimulation improved the final cytotoxic activity in only half of the assays (16/31). When significantly enhanced (>10%, n=8), responder cells resulted in a final 43% cytotoxicity against autologous RCC cells. Mechanism of lysis was at least in part class I mediated. Effector cells have no lytic activity against normal renal cells. Percentage of cells with regulatory T-cell phenotype was not found to be enhanced in the DC-Tu stimulated lymphocytes. Individual differences were observed in the characteristics of DCs generated from RCC patients in contrast to that observed in donors and could explain why lymphocyte stimulation was not improved by DC-Tu in half of the RCC assays. T-cell spreading was suitable for a therapeutic use (>10(9) cells) irrespective of the procedure (with or without DC-Tu stimulation) or the tissular origin of lymphocytes from patients. Data show that precursors of selective antitumor effector cells are present in patients with RCC and can be amplified in vitro either with or without DC-Tu stimulation. One of these populations could be chosen for an adoptive transfer immunotherapy.

Interleukin-6 and vascular endothelial growth factor release by renal cell carcinoma cells impedes lymphocyte–dendritic cell cross-talk

“…Tumour biopsies were minced into small pieces and cultured for a short time (< four passages) in RPMI-1640 medium (Eurobio, Suresnes, France) containing 10% FCS. Tumour cells were then recovered and treated with either 75 grays irradiation (Tu irr) [22] or with 1 mM hydrogen peroxide (H2O2, Sigma, Saint Quentin Fallavier, France) for 12 h (Tu H2O2) [13]. H2O2 treatment could be combined with heat shock for 30 min at 44°C (Tu H2O2 + HS).…”

Dendritic cells incubated with irradiated tumor cells effectively stimulate T lymphocyte activation and induce enhanced expression of CD69, CD25 as well as production of IFNγ and IL4

“…The result was an increased number of IFNg producing cells, as well as the increased IFNa and IL15 concentrations in the supernatants [17]. However, the experience of various tumor immunologists is that the whole cell vaccines stimulate the immune system more efficiently than the oncolysates and that the tumor cell membrane integrity is important for the CTL activation by the tumor vaccines [10,[18][19][20]. In this line of work are the studies of Galea-Lauri et al and Parkhurst et al Galea-Lauri et al demonstrated that the wholly irradiated tumor cells competently provided the specific stimuli to DC and made them efficient for the activation of T cells [19].…”

Dendritic cells are defective in breast cancer patients: a potential role for polyamine in this immunodeficiency

“…Cytotoxic T lymphocytes (CTLs) directed against tumour cells can be amplified in vitro with the use of DCs pulsed with treated tumour cells (DC-Tu) [6]. When assays were done with cells from healthy donors, DC-Tu stimulation repeatedly increased the final cytolytic activity of T cells more than twofold.…”

“…When assays were done with cells from healthy donors, DC-Tu stimulation repeatedly increased the final cytolytic activity of T cells more than twofold. However, we observed that a similar procedure applied to cells from cancer patients enhanced the final cytotoxic activity against autologous tumour only in half of the assays [6]. We noticed in these experiments that the final yield and phenotype of blood-derived myeloid immature DCs was heterogeneous in cancer patients [6].…”

“…However, we observed that a similar procedure applied to cells from cancer patients enhanced the final cytotoxic activity against autologous tumour only in half of the assays [6]. We noticed in these experiments that the final yield and phenotype of blood-derived myeloid immature DCs was heterogeneous in cancer patients [6]. These findings could be related to a relationship between immune suppression instilled during tumour development, as previously described by Kusmartsev and Gabrilovich [7], and increased production of immature myeloid cells in patients with advanced cancers [8].…”