Three-day dendritic cells for vaccine development: Antigen uptake, processing and presentation

<p>Abstract</p> <p>Background</p> <p>Antigen-loaded dendritic cells (DC) are capable of priming naïve T cells and therefore represent an attractive adjuvant for vaccine development in anti-tumor immunotherapy. Numerous protocols have been described to date using different maturation cocktails and time periods for the induction of mature DC (mDC) <it>in vitro</it>. For clinical application, the use of mDC that can be generated in only three days saves on the costs of cytokines needed for large scale vaccine cell production and provides a method to produce cells within a standard work-week schedule in a GMP facility.</p> <p>Methods</p> <p>In this study, we addressed the properties of antigen uptake, processing and presentation by monocyte-derived DC prepared in three days (3d mDC) compared with conventional DC prepared in seven days (7d mDC), which represent the most common form of DC used for vaccines to date.</p> <p>Results</p> <p>Although they showed a reduced capacity for spontaneous antigen uptake, 3d mDC displayed higher capacity for stimulation of T cells after loading with an extended synthetic peptide that requires processing for MHC binding, indicating they were more efficient at antigen processing than 7d DC. We found, however, that 3d DC were less efficient at expressing protein after introduction of <it>in vitro </it>transcribed (<it>ivt</it>)RNA by electroporation, based on published procedures. This deficit was overcome by altering electroporation parameters, which led to improved protein expression and capacity for T cell stimulation using low amounts of <it>ivt</it>RNA.</p> <p>Conclusions</p> <p>This new procedure allows 3d mDC to replace 7d mDC for use in DC-based vaccines that utilize long peptides, proteins or <it>ivt</it>RNA as sources of specific antigen.</p

Family-Based Model Checking with mCRL2

\u3cp\u3eFamily-based model checking targets the simultaneous verfication of multiple system variants, a technique to handle feature-based variability that is intrinsic to software product lines (SPLs). We present an approach for family-based verification based on the feature μ-calculus μL\u3csub\u3ef\u3c/sub\u3e, which combines modalities with feature expressions. This logic is interpreted over featured transition systems, a well-accepted model of SPLs, which allows one to reason over the collective behavior of a number of variants (a family of products). Via an embedding into the modal μ-calculus with data, underpinned by the general-purpose mCRL2 toolset, off-the-shelf tool support for μLf becomes readily available. We illustrate the feasibility of our approach on an SPL benchmark model and show the runtime improvement that family-based model checking with mCRL2 offers with respect to model checking the benchmark product-by-product.\u3c/p\u3

T-Cells Expressing a Highly Potent PRAME-Specific T-Cell Receptor in Combination with a Chimeric PD1-41BB Co-Stimulatory Receptor Show a Favorable Preclinical Safety Profile and Strong Anti-Tumor Reactivity

The hostile tumor microenvironment (TME) is a major challenge for the treatment of solid tumors with T-cell receptor (TCR)-modified T-cells (TCR-Ts), as it negatively influences T-cell efficacy, fitness, and persistence. These negative influences are caused, among others, by the inhibitory checkpoint PD-1/PD-L1 axis. The Preferentially Expressed Antigen in Melanoma (PRAME) is a highly relevant cancer/testis antigen for TCR-T immunotherapy due to broad expression in multiple solid cancer indications. A TCR with high specificity and sensitivity for PRAME was isolated from non-tolerized T-cell repertoires and introduced into T-cells alongside a chimeric PD1-41BB receptor, consisting of the natural extracellular domain of PD-1 and the intracellular signaling domain of 4-1BB, turning an inhibitory pathway into a T-cell co-stimulatory pathway. The addition of PD1-41BB to CD8+ T-cells expressing the transgenic PRAME-TCR enhanced IFN-γ secretion, improved cytotoxic capacity, and prevented exhaustion upon repetitive re-challenge with tumor cells in vitro without altering the in vitro safety profile. Furthermore, a single dose of TCR-Ts co-expressing PD1-41BB was sufficient to clear a hard-to-treat melanoma xenograft in a mouse model, whereas TCR-Ts without PD1-41BB could not eradicate the PD-L1-positive tumors. This cutting-edge strategy supports development efforts to provide more effective TCR-T immunotherapies for the treatment of solid tumors

Design, Objects and Memory: A Sustainability-Oriented Project Itinerary

Game kasino Concerns related to sustainability are recurrent in Design. Questioning project practices, reassessing principles which are intrinsic to the design process, promoting the dissemination of sustainability-oriented actions, and contributing with a new awareness-raising process about consumption are attitudes that might help in the inclusion of environmental and social requirements in design practices. The relationship between sustainability and esthetic, symbolic and affective aspects is under explored in project elaboration. The project process that emphasizes subjective and material characteristics of objects with a view towards the appreciation of affective memories, through the analysis of users’ real-life experiences, might allow the designer to rethink time, tradition, aesthetics and perception to comprehend characteristics at play that are emotionally sustainable. This paper presents and discusses the ongoing research, whose main objective is to design a sustainability-oriented project itinerary, through affections and memory embedded into the relationship between people and objects

Design Processes and Anticipation

Introducing the cultural and historical background of the relationship between design and anticipation is the aim of this chapter. It first provides an overview of the topic describing the various phases and connotations of this connection in design evolution. Then it discusses the creation of a culture of anticipation in contemporary European, and in particular Italian, design as it has been documented in historical surveys and other mediating channels such as exhibitions. In these works, the narration of the future-focused dimension of the project \u2013 in the form of utopias, fictions, visions, prototypes, and studies \u2013 represents one of the constant interpretations underlying design affirmation and development. In order to outline a complete overview, the third paragraph introduces the approaches of other industrialized countries such the Americas, where the activity of design reflects the contextual factors resulting from its historical origins, not simply from the productive point of view but also as the declaration of the specific identity of each nation. During the first half of the Twentieth century, the perception and planning of the future on the continent were constructed under the influence of the events which gave rise to a space incorporating design as a projectual discipline. The idea of territorial development through foreign references allowed for the evolution of diverse bases for distinct initiatives. This paragraph explains how, from that point in time to the present day, the practice of design in America has become legitimized, with a vision oriented toward the search for innovation opportunities and encompassing both the approach and management of environmental complexity and its accompanying variables. Finally, the last paragraph provides a specific focus into the contemporary debate on the relationship between design and anticipation and outlines a conclusive reflection about: how design processes become today relevant in organizations and develop into ideal spaces for multidisciplinary collaboration in fields with high degrees of uncertainty; how they allow the construction of innovational competencies within these organizations; and, finally, how the label advanced design (the design culture most oriented toward the future) appeared as an anticipatory activity