The era of bioengineering: how will this affect the next generation of cancer immunotherapy?

Immunotherapy consists of activating the patient's immune system to fight cancer and has the great potential of preventing future relapses thanks to immunological memory. A great variety of strategies have emerged to harness the immune system against tumors, from the administration of immunomodulatory agents that activate immune cells, to therapeutic vaccines or infusion of previously activated cancer-specific T cells. However, despite great recent progress many difficulties still remain, which prevent the widespread use of immunotherapy. Some of these limitations include: systemic toxicity, weak immune cellular responses or persistence over time and most ultimately costly and time-consuming procedures. Synthetic and natural biomaterials hold great potential to address these hurdles providing biocompatible systems capable of targeted local delivery, co-delivery, and controlled and/or sustained release. In this review we discuss some of the bioengineered solutions and approaches developed so far and how biomaterials can be further implemented to help and shape the future of cancer immunotherapy. The bioengineering strategies here presented constitute a powerful toolkit to develop safe and successful novel cancer immunotherapies

A cancer vaccine with dendritic cells differentiated with GM-CSF and IFNα and pulsed with a squaric acid treated cell lysate improves T cell priming and tumor growth control in a mouse model.

<i> <b>Introduction:</b> </i> Ovarian cancer is one of the most lethal gynecologic cancers. Relapses after remission are common, hence novel strategies are urgently needed. Our group has previously developed a vaccination approach based on dendritic cells pulsed with HOCl-oxidized tumor lysates. Here we investigate the improvement of this vaccine strategy using squaric acid treatment of cancer cells during tumor lysate preparation and by differentiating dendritic cells in the presence of GM-CSF and IFNα. <i> <b>Methods:</b> </i> Induction of cell death by squaric acid treatment was assessed with propidium iodide (PI) and Annexin V in ID8 tumor cells. High mobility group box 1 (HMGB1) immunogenic status was analyzed using a western blot-based method, as previously described. For immunological tests, ID8 cells expressing ovalbumin (ova-ID8) were treated with squaric acid before cell lysis. DCs prepared with the canonical GM-CSF and IL-4 differentiation cocktail or IFNα and GM-CSF were pulsed with tumor cell lysates and further matured in the presence of IFNγ and LPS (4-DCs and α-DCs respectively). DCs were then used in co-culture assays with ova-specific T cells and IFNγ and IL-4 secretion measured by ELISA. DC phenotypes were characterized by FACS. Finally, DCs were tested in an ovarian cancer mouse model measuring body weight and animal survival. <i> <b>Results:</b> </i> Squaric acid treatment of mouse ovarian cancer cells induced tumor cell death as well as preserve HMGB1, a crucial Damage-associated molecular pattern (DAMP) signal, in its active reduced form. Squaric acid treatment of ID8-ova cells increased IFNγ and decreased IL-4 production from ova-specific T cells in co-culture experiments, promoting a more immunogenic cytokine secretion pattern. DCs differentiated in the presence of IFNα induced a considerable decrease in IL-4 production compared to canonical 4-DCs, without affecting IFNγ release. DC phenotyping demonstrated a more mature and immunogenic phenotype for IFNα-differentiated DCs. Vaccination in tumor-bearing mice showed that IFNα-differentiated DCs pulsed with squaric acid-treated lysates were the most potent at delaying tumor growth, improving animal survival. <i> <b>Conclusion:</b> </i> We identified squaric acid as a novel immunogenic treatment of tumor cells for cancer vaccines particularly efficient in prolonging animal survival when used in combination with IFNα-differentiated DCs. These promising results support future efforts for the clinical translation of this approach

Caratteristiche geomorfologiche dell’Isola di Pianosa.

Pianosa Island belongs to the Tuscany Archipelago; it has an extension of 10 km2, 18 km of coastal development and a max altitude of 29 m a.s.l. The island is made up of neogenic and quaternary sediments; pelitic Miocene sediments outcrops at the base of the cliff in the south-western coast, while the bio-calcarenitic Pliocene sediments cover most part of the island. Quaternary deposits are established by biodetrital shore sediments of Tyrrhenian age and by Late Pleistocene colluvial deposits. The island is characterized by cliffs in the western and southern sector, by plain surfaces and by a small beach in the east side. Cliffs have been distinguished in 3 different types. In the first type is always present a developing wave notch. When the coast is not characterized by cliff we can found erosion plains (at maximum level of approximately 4 m) formed during the Tyrrhenian high stand (Isotopic stage 5e). These erosion planes can be suspended and sub-horizontal (when the layers of the pliocenic substrate dip towards inland), or gently tilted towards sea (when the layers of the pliocenic substrate dip towards sea). On these various erosion planes are always present rock pools characterized by different shape and dimension, besides rare fossiliferous shore deposits (Panchina). In these sediments have been recognized, between others: Strombus bubonius, Patella ferruginea and Conus testudinarius, dating Tyrrhenian age. On the island we can found three different order of erosion planes at 2-4 m, 8 m and 25 m a.s.l. Karst process produced erosion epigean micro-landforms, such us dissolutional flutes, dissolutional groves and dissolutional pans (kamenitza), macro-landforms represented by E-W karst oval basin depressions, enlarged hundred meters and few meters deep. The same process left isolated rock promontory residual of the tyrrhenian cliffs coast lines. Much diffused are hypogean karst shape such as caves and conduits that, sometimes, are opened on the cliff and are visible only by the sea. In some of these caves, characterized by speleothemes, we discovered, at a quote of 7 m a.s.l., marine deposits and litodomes holes probably of Tyrrhenian age. Frequently is the founding of breccias breeded by collapse of caves and conduits. Surface karst landforms are widening covered by high agricoltural activity developed in the last two centuries due to the presence of the agricoltural penal colony

Abnormalities of structure and function in the neuromuscular junction of dystrophic (mdx) mice

pubblicato su :J. Musc. Cell Mot. 21(8):791-833 (2000

Determination of high-energy phosphate compounds and inorganic phosphate by reversed-phase high-performance liquid chromatography: evaluation of myocardial metabolic status in aerobically perfused and hypoxic mouse heart.

The present paper describes a simple HPLC method designed for measuring high-energy phosphate (HEP) compounds in a single run and inorganic phosphate (Pi) in an other short run under the same HPLC conditions. Inorganic phosphate was estimated by using thymidine phosphorylase (EC 2.4.2.4) which catalyzes a reaction involving inorganic phosphate to produce 2-deoxyribose 1-phosphate and thymine. The thymine/Pi stoichiometry was 1. The method provides a reproducible instrument for evaluating myocardial high-energy metabolism under physiological and pathological conditions