Engineered Cell-Based Therapeutics: Synthetic Biology Meets Immunology

Synthetic Biology has enabled new approaches to several medical applications including the development of immunotherapies based on bioengineered cells, and most notably the engineering of T-cells with tumor-targeting receptors, the Chimeric Antigen Receptor (CAR)-T cells. CAR-T-cells have successfully treated blood tumors such as large B-cell lymphoma and promise a new scenario of therapeutic interventions also for solid tumors. Learning the lesson from CAR-T cells, we can foster the reprogramming of T lymphocytes with enhanced survival and functional activity in depressing tumor microenvironment, or to challenge diseases such as infections, autoimmune and chronic inflammatory disorders. This review will focus on the most updated bioengineering approaches to increase control, and safety of T-cell activity and to immunomodulate the extracellular microenvironment to augment immune responses. We will also discuss on applications beyond cancer treatment with implications toward the understanding and cure of a broader range of diseases by means of mammalian cells engineering

Novel Immunoregulatory Functions of IL-18, an Accomplice of TGF-\u3b21

TGF-\u3b21 is a pleiotropic factor exerting a strong regulatory role in several cell types, including immune cells. In NK cells it profoundly alters the surface expression of crucial activating and chemokine receptors. To understand which soluble signals might better contrast these effects, we cultured human NK cells in the presence of TGF-\u3b21 and different innate and adaptive cytokines, generally referred as "immunostimulatory". These included IL-2, IL-15, IL-21, IL-27, and IL-18. Unexpectedly, IL-18 strengthened rather than contrasting important TGF-\u3b21-mediated functions. In particular, IL-18 further reduced the expression of CX\u2083CR1 and NKp30, leading to the virtual abrogation of the triggering capability of this activating receptor. Moreover, IL-18 further increased the expression of CXCR4. The IL-18-mediated additive effect on NKp30 and CXCR4 expression involved transcriptional regulation and activation of MEK/ERK and/or p38MAPK. A proteomic approach quantified both surface and intracellular proteins significantly modified in cytokine-treated NK cells, thus giving global information on the biological processes involving TGF-\u3b21 and IL-18. Our data support the concept that IL-18 may have a different behavior depending on the type of soluble factors characterizing the microenvironment. In a TGF-\u3b21 rich milieu such as tumors, it may contribute to the impairment of both NK cells recruitment and killing capability

TGF- f1-Mediated Regulation of the Human NK miRNome in the Tumor Microenvironment

Regolazione Mediata da TGF- f1 del miRNoma di Cellule NK Umane nel Microambiente Tumoral

Lamb Waves Propagation along 3C-SiC/AlN Membranes for Application in Temperature-Compensated, High-Sensitivity Gravimetric Sensors

The propagation of the fundamental quasi-symmetric Lamb mode S0 travelling along 3C-SiC/c-AlN composite plates is theoretically studied with respect to the AlN and SiC film thickness, the acoustic wave propagation direction and the electrical boundary conditions. The temperature effects on the phase velocity have been considered for four AlN/SiC-based electroacoustic coupling configurations, specifically addressing the design of temperature-compensated, enhanced-coupling, GHz-range electroacoustic devices. The gravimetric sensitivity and resolution of the four temperature-stable SiC/AlN composite structures are theoretically investigated with respect to both the AlN and SiC sensing surface. The SiC/AlN-based sensor performances are compared to those of surface acoustic waves and Lamb S0 mode mass sensors implemented on bulk conventional piezoelectric materials and on thin suspended membranes

A Bayesian bivariate hierarchical model with correlated parameters for the analysis of road crashes in Italian tunnels

An analysis of crashes occurring in 252 unidirectional Italian motorway tunnels over a 4-year monitoring period is provided to identify the main causes of crashes in tunnels. In this paper, we propose a full Bayesian bivariate Poisson lognormal hierarchical model with correlated parameters for the joint analysis of crashes of two levels of severity, namely severe (including fatality and injury accidents only) and non-severe (property damage only), providing better insight on the available data with respect to an analysis based on severe and non-severe independent univariate models. In particular, the proposed model shows that for both of severity levels the crash frequency increases with some parameters: the average annual daily traffic per lane, the tunnel length, and the percentage of trucks, while the presence of the sidewalk provides a reduction in severe accidents. Also the presence of the third lane induces a reduction in severe accidents. Moreover, a reduction in the crash frequency of the two crash-types over years is present. The correlation between the parameters might offer additional insights into how some combinations can affect safety in tunnels. The results are critically discussed by highlighting strength and weakness of the proposed methodology

Main NK cell receptors and their ligands: regulation by microRNAs

The NK cells functions are finely tuned by several kinds of inhibitory and activating receptors, whose pattern of expression characterizes different NK subpopulations and varies with the cell activation status. MicroRNAs have an important role in tightly regulating the expression of NK receptors and, analogously, the expression of their ligands in target cells. The relevance of the microRNA-mediated control is highlighted by the dysregulation of these pathways observed in cancer and virus-infected cells. Here we review our current knowledge of the microRNAs involved in the regulation of NK receptors, as well as that of the corresponding cellular ligands

miR-24-3p down-regulates the expression of the apoptotic factors FasL and BIM in human natural killer cells

MicroRNAs are involved in the regulation of different functions in immune and non-immune cells. Here we show that miR-24-3p functionally interacts with FASLG mRNA and down-regulates its expression. This interaction occurs in human natural killer cells (NK), leading to the modulation of FasL surface expression. Moreover, miR-24-3p also modulates the mRNA and protein expression of BIM in NK cells. Thus, it likely contributes to the control of both the extrinsic and intrinsic apoptotic pathways. In line with this hypothesis, inhibition of miR-24-3p improves both initiator caspase-8 and effector caspase-3 and -7 activities, increases cell apoptosis, and reduces cell viability. Our data suggest that miR-24-3p can act as a survival factor in NK cells, affecting the FasL-mediated killing of Fas expressing cells and the BIM-dependent cell death. More generally, miR-24-3p may condition the level of cell apoptosis, which increases at the contraction phase of the immune response when the clearance of various expanded effector cells is needed

MRI findings in patient with primary cutaneous nocardiosis: A case report

: Nocardia is a genus of gram-positive, filamentous, aerobic bacteria that belongs to the Actinomycetales order. With over 50 species, it is ubiquitous in dust, soil, decaying organic matter, and stagnant water. Inhalation of the pathogen often leads to pulmonary nocardiosis, while extrapulmonary nocardiosis can affect the central nervous system, skin, and subcutaneous tissues. Primary cutaneous nocardiosis occurs when the pathogen is introduced through a skin lesion or insect bite, for example, this report presents a case of primary cutaneous nocardiosis in a patient with Minimal Change Glomerulonephritis and iatrogenic immunosuppression. Magnetic resonance imaging revealed extensive involvement of the skin, subcutaneous tissue, and lower limb muscles

TGF-beta1 downregulates the expression of CX3CR1 by inducing miR-27a-5p in primary human NK cells

Activity of human natural killer (NK) cells against cancer cells is deeply suppressed by TGF-beta1, an immunomodulatory cytokine that is released and activated in the tumor microenvironment. Moreover, our previous data showed that TGF-beta1 modifies the chemokine receptor repertoire of NK cells. In particular, it decreases the expression of CX3CR1 that drives these effectors toward peripheral tissues, including tumor sites. To identify possible mechanisms mediating chemokine receptors modulation, we analyzed the microRNA profile of TGF-beta1-treated primary NK cells. The analysis pointed out miR-27a-5p as a possible modulator of CX3CR1. We demonstrated the functional interaction of miR-27a-5p with the 3' untranslated region (3'UTR) of CX3CR1 mRNA by two different experimental approaches: by the use of a luciferase assay based on a reporter construct containing the CX3CR1 3'UTR and by transfection of primary NK cells with a miR-27a-5p inhibitor. We also showed that the TGF-beta1-mediated increase of miR-27a-5p expression is a consequence of miR-23a-27a-24-2 cluster induction. Moreover, we demonstrated that miR-27a-5p downregulates the surface expression of CX3CR1. Finally, we showed that neuroblastoma cells induced in resting NK cells a downregulation of the CX3CR1 expression that was paralleled by a significant increase of miR-27a-5p expression. Therefore, the present study highlights miR-27a-5p as a pivotal TGF-beta1-induced regulator of CX3CR1 expression