“…To analyze the effect of RIG-I stimulation on formation and function of tumor-EVs, we used the human melanoma cell lines D04mel and Ma-Mel-86c 28,29 . In line with RIG-I as type I Interferon (IFN)-dependent gene, baseline expression of RIG-I in all used cell lines was strongly increased by type I IFN (Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Figure 1.RIG-I stimulation triggers the release of extracellular vesicles (EVs). (A) Melanoma cells D04mel 28 or Ma-Mel-86c 29 were analyzed for mRNA expression of RIG-I by quantitative real-time PCR in the presence or absence of type I Interferon (IFN) stimulation. Data are normalized to β-Actin.…”
Activation of the innate immune receptor retinoic acid-inducible gene I (RIG-I) by its specific ligand 5′-triphosphate-RNA (3pRNA) triggers antitumor immunity predominantly via NK cell activation and direct apoptosis induction in tumor cells. However, how NK cells are mobilized to attack the tumor cells remains elusive. Here, we show that RIG-I activation induced the secretion of extracellular vesicles (EVs) from melanoma cells, which by themselves revealed antitumor activity in vitro and in vivo. RIG-I-induced EVs from melanoma cells exhibited an increased expression of the NKp30-ligand (BAG6, BAT3) on their surface triggering NK cell-mediated lysis of melanoma cells via activation of the cytotoxicity NK cell-receptor NKp30. Moreover, systemic administration of RIG-I-induced melanoma-EVs showed a potent antitumor activity in a melanoma mouse model in vivo. In conclusion, our data establish a new RIG-I-dependent pathway leading to NK cell-mediated tumor cell killing.
“…To analyze the effect of RIG-I stimulation on formation and function of tumor-EVs, we used the human melanoma cell lines D04mel and Ma-Mel-86c 28,29 . In line with RIG-I as type I Interferon (IFN)-dependent gene, baseline expression of RIG-I in all used cell lines was strongly increased by type I IFN (Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Figure 1.RIG-I stimulation triggers the release of extracellular vesicles (EVs). (A) Melanoma cells D04mel 28 or Ma-Mel-86c 29 were analyzed for mRNA expression of RIG-I by quantitative real-time PCR in the presence or absence of type I Interferon (IFN) stimulation. Data are normalized to β-Actin.…”
Activation of the innate immune receptor retinoic acid-inducible gene I (RIG-I) by its specific ligand 5′-triphosphate-RNA (3pRNA) triggers antitumor immunity predominantly via NK cell activation and direct apoptosis induction in tumor cells. However, how NK cells are mobilized to attack the tumor cells remains elusive. Here, we show that RIG-I activation induced the secretion of extracellular vesicles (EVs) from melanoma cells, which by themselves revealed antitumor activity in vitro and in vivo. RIG-I-induced EVs from melanoma cells exhibited an increased expression of the NKp30-ligand (BAG6, BAT3) on their surface triggering NK cell-mediated lysis of melanoma cells via activation of the cytotoxicity NK cell-receptor NKp30. Moreover, systemic administration of RIG-I-induced melanoma-EVs showed a potent antitumor activity in a melanoma mouse model in vivo. In conclusion, our data establish a new RIG-I-dependent pathway leading to NK cell-mediated tumor cell killing.
“…Melanoma cell lines (denoted A02-M and D14-M) were established from metastases from trial participants A02 and D14 [14], [15]. In brief, cells from mechanically disaggregated tumours were cultured in complete medium to establish cell lines, which were confirmed to be of melanoma origin by expression profiling [15], and authenticated via short tandem repeat profiling according to the manufacturer's instructions (AmpF‘STR Profiler Plus ID kit; Applied Biosystems, Foster City, CA, USA).…”
While cloned T cells are valuable tools for the exploration of immune responses against viruses and tumours, current cloning methods do not allow inferences to be made about the function and phenotype of a clone's in vivo precursor, nor can precise cloning efficiencies be calculated. Additionally, there is currently no general method for cloning antigen-specific effector T cells directly from peripheral blood mononuclear cells, without the need for prior expansion in vitro. Here we describe an efficient method for cloning effector T cells ex vivo. Functional T cells are detected using optimised interferon gamma capture following stimulation with viral or tumour cell-derived antigen. In combination with multiple phenotypic markers, single effector T cells are sorted using a flow cytometer directly into multi-well plates, and cloned using standard, non antigen-specific expansion methods. We provide examples of this novel technology to generate antigen-reactive clones from healthy donors using Epstein-Barr virus and cytomegalovirus as representative viral antigen sources, and from two melanoma patients using autologous melanoma cells. Cloning efficiency, clonality, and retention/loss of function are described. Ex vivo effector cell cloning provides a rapid and effective method of deriving antigen-specific T cells clones with traceable in vivo precursor function and phenotype.
“…In addition, dendritic cells have been directly fused with autologous breast cancer tumor cells, which allows for the presentation of multiple tumor antigens (55). Although dendritic cell-based vaccines have had minimal side effects and have induced measurable T-cell immunity, few durable clinical responses have been reported (55–57). However, a dendritic cell-based vaccine was shown to confer a modest survival benefit in hormone-refractory prostate cancer (58).…”
SUMMARY
The goal of cancer vaccines and immunotherapies is to train the immune system to recognize cancer cells and destroy them. Immune responses play a dynamic role in the development of cancers, from immunosurveillance to immune escape; from in situ immune dysregulation to metastatic spread. The systematic identification and targeting of molecules involved in the immune response has led to a wide variety of potential immunotherapeutic targets for the treatment of breast cancer. Extraordinary advances in molecular immunology have led to a detailed understanding of tumor antigens, antigen presentation, innate immunity, cytokine and chemokine pathways, and immunoregulation. Many of these vaccine therapies are already in clinical development. It is the rational and rapid translation of these scientific discoveries into effective therapies for patients with breast cancer that poses the greatest challenge, and opportunity, to realize the potential of tumor vaccine therapy for breast cancer.
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