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Dendritic Cells Primed with Protein–Protein Fusion Adjuvant
To develop efficient T cell priming cancer vacciness, various recombinant fusion proteins have been developed by fusing a tumor antigen with a protein capable of stimulating or targeting dendritic cells (DC), the most important antigen-presenting cells for inducing CD8+ cytotoxic T lymphocytes (CTL) which can efficiently kill tumor cells expressing the tumor antigen. The DC-stimulating or DC-targeting proteins, including granulocyte/macrophage colony-stimulating factor (GM-CSF), anti-DEC-205 monoclonal antibodies, flagellin, and heat shock proteins (HSP), function as promising intermolecular adjuvants. Herein, we describe ...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Tumor Antigen-/Cytokine-Pulsed Dendritic Cells in Therapy Against Lymphoma
Adoptive cell therapy using dendritic cells (DCs) is a strategy to deliver tumor antigens in cancer immunotherapy. Co-delivery of antigens to DC with essential components like genes encoding cytokines, chemokines, and other molecules or stimulation with recombinant cytokines is a potential method for designing an effective tumor vaccines protocol. Here, we describe the stimulation of purified splenic- or bone marrow-derived DC with recombinant interleukin-15 (IL-15) in the presence of intact soluble antigen from metastatic lymphoma tumor cells in an experimental animal model. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Ex Vivo Loading of Autologous Dendritic Cells with Tumor Antigens
We describe the methods to obtain whole antigens from autologous tumor tissues in order to load DC generated ex vivo from patients with gastrointestinal cancer. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Antigen Trapping by Dendritic Cells for Antitumor Therapy
Dendritic cells (DC) are potent antigen-presenting cells (APC) that are capable of stimulating both naive CD4+ T helper cells and CD8+ cytotoxic T cells. Therefore, DC are being extensively evaluated as vehicles for antigen delivery in immunotherapies for the treatment of patients with cancer. Many techniques have been used to load DC with tumor-associated antigens (TAA), including pulsing with synthetic peptides that represent T cell epitopes. This strategy has been used in several human clinical vaccination trials; however, it is limited to patients who express the particular peptide MHC-restricting molecule. Alternative...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Pulsing Dendritic Cells with Whole Tumor Cell Lysates
One of the strategies employed in immunotherapy for cancer is to use of ex vivo-generated dendritic cells (DC) pulsed with tumor antigens. Several approaches have been used to obtain and load tumor antigens in DC. One such technique is to use whole tumor cell lysate from one or more tumor cell lines of the tumor type to be treated. The advantage of applying this method is that it provides a large spectrum of tumor antigens. However, some considerations must be taken into account to obtain a lysate with appropriate biological activity, such as cell line harvest and the method to lyse the cells. In this chapter, we describe ...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Generation of Multiple Peptide Cocktail-Pulsed Dendritic Cells as a Cancer vaccines
Cancer immunotherapy based on dendritic cell (DC) vaccination has promising alternatives for the treatment of cancer. A central tenet of DC-based cancer immunotherapy is the generation of antigen-specific cytotoxic T lymphocyte (CTL) response. Tumor-associated antigens (TAA) and DC play pivotal roles in this process. DCs are well known to be the most potent antigen-presenting cells and have the most powerful antigen-presenting capacity. DCs pulsed with various TAA have been shown to be effective in producing specific antitumor effects both in vitro and in vivo. Several types of tumor antigens have been applied in cancer tr...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Single-Step Antigen Loading and Maturation of Dendritic Cells Through mRNA Electroporation of a Tumor-Associated Antigen and a TriMix of Costimulatory Molecules
Dendritic cells (DC) are key players in several types of cancer vacciness. Large numbers of DC can easily be generated in closed systems from the monocyte fraction of the peripheral blood. They are the professional antigen-presenting cells, and electroporation of mRNA-encoding tumor antigens is a very efficient and a relatively simple way to load the DC with antigen. The co-electroporation of a tumor antigen of choice and the combination of 3 costimulatory molecules, including CD70, caTLR4, and CD40L (TriMix-DC), leads to fully potent antigen-presenting DC able to generate a broad immune response. (Source: Springer protoco...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Exploiting the CD1d-iNKT Cell Axis for Potentiation of DC-Based Cancer vacciness
Invariant natural killer T cells (iNKT) and dendritic cells (DC) play a central role in tumor immunity through downstream activation of immune effector cells by pro-inflammatory cytokines. Evidence is accumulating that the CD1d-iNKT cell axis can be effectively used to potentiate DC-based cancer vacciness. Here, we provide a detailed methodology for the generation of (CD1d-expressing) monocyte-derived DC (moDC) and their subsequent loading with the iNKT cell agonist α-galactosylceramide (α-GalCer) or their direct ligation by agonistic anti-CD1d monoclonal antibodies. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Intratumoral Injection of BCG-CWS-Pretreated Dendritic Cells Following Tumor Cryoablation
Intratumoral administration of dendritic cells (DC) following cryoablation of tumor is one of the personalized cancer immunotherapies which is able to induce immune responses to multiple endogenous tumor antigens, including shared and unique antigens. Here we describe protocols of cryoablation of tumors, generation of cultured DC, pretreatment of DC with a Toll-like receptor (TLR)-stimulating purified component of Bacillus Calmette-Guerin cell wall fraction (BCG-CWS) and highly immunogenic keyhole limpet hemocyanin (KLH) antigen, and combined use of tumor cryoablation and intratumoral administration of BCG-CWS-pretreated D...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Fast Monocyte-Derived Dendritic Cell-Based Immunotherapy
Recent reports have described a new strategy for differentiation and maturation of monocyte (Mo)-derived dendritic cells (DC) within only 48–72 h of in vitro culture (fast-DC). Mature fast-DC are as effective as mature standard-DC (generated in 7–10 days of in vitro culture) in priming and propagation of antigen-specific T-cell responses. The use of fast-DC not only reduces labor and supply cost, as well as workload and time, but also increases the DC yield from Mo, which may facilitate DC-based immunotherapy for cancer patients. Detailed protocols for generation, pulsing with different antigen sources, and tra...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Genetic Modification of Dendritic Cells with RNAi
Gene silencing with RNAi is an invaluable technique in cell biology to knock down the target gene expression. Dendritic cells (DC) are the most effective antigen-presenting cells (APC), and the efficacy of antigen presentation is tightly controlled by the stimulatory as well as inhibitory mechanisms. In recent studies, RNAi technology has been employed to silence the expression of the intrinsic inhibitors of antigen presentation in DC, improving the efficacy of DC vacciness against tumor antigens in pre-clinical studies. Here, we describe the technique of using siRNA oligonucleotides, adenovirus expressing shRNA (Ad-shRNA)...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Dendritic Cells Transfected with Adenoviral Vectors as vacciness
Dendritic cells (DCs) are critical to the initiation of a T-cell response. They constitute the most potent antigen-presenting cell (APC) endowed with the unique capacity to stimulate an antigen-specific T-cell responses by naïve T cells. Adenoviruses (Ad) have high transduction efficiency for many cell types including cells of hematopoietic origin independent of their mitotic status, and replication-defective Ad have demonstrated a safety profile clinically. Further, Ad vectors provide a high level of transgene expression, and Ad-transduced DCs can effectively present antigenic proteins. In this chapter, we outline a ...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news

Erratum
(Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - December 5, 2013 Category: Cancer & Oncology Source Type: news

Markers for Anti-cytotoxic T-lymphocyte Antigen 4 (CTLA-4) Therapy in Melanoma
Therapeutic strategies that block Cytotoxic T lymphocyte antigen-4 (CTLA-4) enhance antitumor immunity and prolong the lives of patients with metastatic melanoma. However, only a subset of patients benefit, and responses are often delayed due to heterogeneous response kinetics. Ongoing monitoring of the immunologic effects of therapy and correlating these immunologic changes with patient outcomes continue to be important goals to better identify possible mechanisms of clinical activity of these agents. This chapter introduces the major areas of investigation in monitoring patients treated with CTLA-4 blockade and provides ...
Source: Springer protocols feed by Cancer Research - November 27, 2013 Category: Cancer & Oncology Source Type: news

Immunologic Monitoring of Cancer Vaccine Trials Using the ELISPOT Assay
We present the Interferon (IFN)-γ-producing lymphocyte assay, but the platform is easily adjusted to several cell types and several secreted molecules. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 27, 2013 Category: Cancer & Oncology Source Type: news