Springer protocols feed by Cancer Research 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Using LacO Arrays to Monitor DNA Double-Strand Break Dynamics in Live Schizosaccharomyces pombe Cells
LacO arrays, when combined with LacI-GFP, have been a valuable tool for studying nuclear architecture and chromatin dynamics. Here, we outline an experimental approach to employ the LacO/LacI-GFP system in S. pombe to assess DNA double-strand break (DSB) dynamics and the contribution of chromatin state to DSB repair. Previously, integration of long, highly repetitive LacO arrays in S. pombe has been a challenge. To address this problem, we have developed a novel approach, based on the principles used for homologous recombination-based genome engineering in higher eukaryotes, to integrate long, repetitive LacO arrays with t...
Source: Springer protocols feed by Cancer Research - July 17, 2014 Category: Cancer & Oncology Source Type: news
Arginine-Grafted Biodegradable Polymer: A Versatile Transfection Reagent for both DNA and siRNA
Effective delivery of DNA or siRNA into primary cells demands an efficient delivery system. However, the significant differences in physical and molecular characteristics of the two molecules generally necessitate distinct delivery systems or considerable differences in carrier formulation protocols for effective transfection. Arginine-grafted bioreducible poly (disulfide amine) (ABP) is a redox-sensitive, bioreducible, positively charged polymer which complexes with siRNA and DNA via charge interactions to form nanoplexes. ABP effectively mediates cytoplasmic delivery of both DNA and siRNA into multiple cell types, includ...
Source: Springer protocols feed by Cancer Research - July 17, 2014 Category: Cancer & Oncology Source Type: news
Heat Shock Proteins Purified from Autologous Tumors Using Antibody-Based Affinity Chromatography
Heat shock proteins (HSP) isolated from autologous tumors have become a promising tool for active-specific anticancer immunotherapy due to their properties as carriers of antigenic peptides on one hand and as immunostimulatory adjuvants on the other. Rapid and efficient isolation of HSP-peptide complexes from a patient’s tumor is fundamental for their clinical application. Herein, we describe the purification of the HSP Gp96 and Hsc70/Hsp70 from human autologous tumor sources by one-step antibody-based affinity chromatography. Recombinant anti-Gp96 and anti-Hsp70 single-chain Fv antibodies are covalently coupled to a...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Cloning Variable Region Genes of Clonal Lymphoma Immunoglobulin for Generating Patient-Specific Idiotype DNA vaccines
Available therapies for lymphoplasmacytic lymphoma (LPL) provide no survival advantage if started before signs or symptoms of end-organ damage develop; hence, current recommendations are to follow a program of observation while patients are in the asymptomatic phase of disease. We hypothesize that using idiotypic determinants of a B-cell lymphoma’s surface immunoglobulin as a tumor-specific marker, we can develop patient-specific chemokine-idiotype fusion DNA vacciness that induce an immune response against LPL. By activating the host immune system against the tumor antigen, we postulate that disease control of asymp...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
The Direct Display of Costimulatory Proteins on Tumor Cells as a Means of Vaccination for Cancer Immunotherapy
Therapeutic vacciness against cancer are at their prime, owing to our comprehensive understanding of immune effector responses generated against tumor and the mechanisms employed by the progressing tumor to evade the immune system. The immune system is primed by tumor-associated antigens (TAA) that are perceived as foreign. Therefore, the identification of TAA led to the development of subunit vaccines formulations comprising defined TAA as stand-alone vacciness or in combination with immune adjuvants. Inasmuch as cancer cells express a diverse set of TAA, novel immunomodulatory approaches that not only use tumor cells as ...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Tumor Cell Transformation Using Antisense Oligonucleotide
Major histocompatibility complex (MHC) Class II-positive, invariant chain (Ii)-suppressed tumor cells induce both T helper and cytotoxic T lymphocytes’ responses. Genetically controlled immunotherapy could be utilized for prophylactic vaccination of tumor-free individuals who are at high risk of developing tumor and can be therapeutic for treating established tumors that are nonresponsive to existing therapies. In this chapter, we provide practical methods to create a potent in vivo tumor cell vaccines by inducing MHC Class II and Ii using MHC Class II transactivator (CIITA) or interferon-gamma (IFN-γ) and subs...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Autologous Tumor Cells Engineered to Express Bacterial Antigens
Cancer immunotherapies are emerging as promising treatment modalities in the management of the disease. As a result, cancer vacciness are considered to be immensely crucial in preventing recurrence, a well-known nemesis in cancer patients because they have the potential to activate memory antitumor immunity. Due to poor antigenicity and self-tolerance, most tumor antigens require interventional vaccines therapies to provide an adequate “danger” signal to the immune system in order to activate a robust, clinically meaningful antitumor immunity. It has been postulated that this requirement may be achieved by prov...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Loading of Acute Myeloid Leukemia Cells with Poly(I:C) by Electroporation
In this chapter, we describe the technique of electroporation as an efficient method to load primary leukemic cells with the double-stranded RNA (dsRNA) analogue, polyriboinosinic polyribocytidylic acid (poly(I:C)), and detail on the delicate freezing and thawing procedure of primary leukemic cells. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Cytokine Activation of Natural Killer Cells
We describe in this chapter the method to activate NK cells with cytokines and to ascertain their degree of activation. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Monitoring the Frequency and Function of Regulatory T Cells and Summary of the Approaches Currently Used to Inhibit Regulatory T Cells in Cancer Patients
Regulatory T cells (Treg) are a subset of T lymphocytes that in humans represent less than the 10 % of circulating CD4+ T cells. Treg are specialized in the inhibition of the immune responses and play a crucial role in the maintenance of immunological tolerance. Several lines of evidence clearly documented the role of Treg in restraining antitumor immune responses. For this reason, antitumor immunotherapy approaches have been recently associated with drug treatments aimed at depleting Treg or blocking their functions. A summary of the currently used in vivo approaches to limit Treg expansion in cancer patients is here prov...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Genetic Modification of Cytotoxic T Lymphocytes to Express Cytokine Receptors
Adoptive transfer of tumor-infiltrating lymphocytes (TIL) or antigen-specific cytotoxic T lymphocytes (CTL) is safe and can be effective in cancer patients. Achievement of clinical responses in these patients is associated with the in vivo expansion and persistence of the transferred T lymphocytes. For this reason, recombinant human interleukin-2 (IL-2) is frequently used to support the in vivo survival of T lymphocytes infused into patients. However, IL-2 also causes important side effects. Thus, alternative strategies are highly demanded to limit cytokine-related off-target effects and to redirect the responsiveness of s...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Genetic Modification of Mouse Effector and Helper T Lymphocytes Expressing a Chimeric Antigen Receptor
Genetic modification of primary mouse T cells with chimeric antigen receptors (CAR) has emerged as an important tool for optimizing adoptive T cell immunotherapy strategies. However, limitations in current protocols for generating highly pure and sufficient numbers of enriched effector and helper CAR+ T cell subsets remain problematic. Here, we describe a new retroviral transduction protocol for successfully generating transduced CD8+ and CD4+ T lymphocytes for in vitro and in vivo characterization. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Modification of T Lymphocytes to Express Tumor Antigens
Human T cells can be genetically modified to express tumor-associated antigens (TAA) for the induction of tumor-specific immunity, suggesting that T cells may be alternative candidates of effective antigen-presenting cells (TAPC) and may be useful in vivo as cellular cancer vacciness. The effective induction of TAA-specific T cell immune responses requires activation of T cells by CD3/CD28 antibodies and the presence of proinflammatory cytokines such as interleukin-7 (IL-7) and interleukin-12 (IL-12). Here, we describe the technique of preparing activated human TAPC pulsed with TAA peptides for the induction of tumor antig...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Electroporation of Dendritic Cells with Autologous Total RNA from Tumor Material
Dendritic cells (DC) are unique antigen-presenting cells that initiate and orchestrate adaptive immunity. Theoretically, cancer cells that express tumor-specific antigens can be destroyed by cytotoxic T cells. However, inherent antitumor responses are often not efficient, since tumor cells can mask their antigens and do not activate DC, an event required for the development of tumor antigen-specific cytotoxic T cell responses. Over a decade ago, the ex vivo preparation of autologous tumor antigen-loaded monocyte-derived DC vacciness as a novel potent anticancer strategy was launched. Phase I and II trials have been perform...
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
Antigen-Specific mRNA Transfection of Autologous Dendritic Cells
Several reports have described the use of tumor-extracted RNA as source of tumor antigen for the preparation of vacciness based on dendritic cells (DC) and its potential use for antigen-specific or polyvalent tumor vaccination. Upon transfection, RNA is transcribed into proteins that enter the cytoplasmic degradation pathway and can be presented by DC through class I major histocompatibility complex (MHC)-I, thus inducing specific T cell cytotoxic responses. In this chapter, we present a protocol to transfect murine dendritic cells with tumor mRNA by means of electroporation. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - April 7, 2014 Category: Cancer & Oncology Source Type: news
