In Vitro Modeling of Tumor-Immune System Interaction.

Immunotherapy has emerged during the past two decades as an innovative and successful form of cancer treatment. However, frequently, mechanisms of actions are still unclear, predictive markers are insufficiently characterized, and preclinical assays for innovative treatments are poorly reliable. In this context, the analysis of tumor/immune system interaction plays key roles, but may be unreliably mirrored by in vivo experimental models and standard bidimensional culture systems. Tridimensional cultures of tumor cells have been developed to bridge the gap between in vitro and in vivo systems. Interestingly, defined aspects of the interaction of cells from adaptive and innate immune systems and tumor cells may also be mirrored by 3D cultures. Here we review in vitro models of cancer/immune cell interaction and we propose that updated technologies might help develop innovative treatments, identify biologicals of potential clinical relevance, and select patients eligible for immunotherapy treatments

High myeloperoxidase positive cell infiltration in colorectal cancer is an independent favorable prognostic factor

BACKGROUND Colorectal cancer (CRC) infiltration by adaptive immune system cells correlates with favorable prognosis. The role of the innate immune system is still debated. Here we addressed the prognostic impact of CRC infiltration by neutrophil granulocytes (NG). METHODS A TMA including healthy mucosa and clinically annotated CRC specimens (n = 1491) was stained with MPO and CD15 specific antibodies. MPO+ and CD15+ positive immune cells were counted by three independent observers. Phenotypic profiles of CRC infiltrating MPO+ and CD15+ cells were validated by flow cytometry on cell suspensions derived from enzymatically digested surgical specimens. Survival analysis was performed by splitting randomized data in training and validation subsets. RESULTS MPO+ and CD15+ cell infiltration were significantly correlated (p<0.0001; r = 0.76). However, only high density of MPO+ cell infiltration was associated with significantly improved survival in training (P = 0.038) and validation (P = 0.002) sets. In multivariate analysis including T and N stage, vascular invasion, tumor border configuration and microsatellite instability status, MPO+ cell infiltration proved an independent prognostic marker overall (P = 0.004; HR = 0.65; CI:±0.15) and in both training (P = 0.048) and validation (P = 0.036) sets. Flow-cytometry analysis of CRC cell suspensions derived from clinical specimens showed that while MPO+ cells were largely CD15+/CD66b+, sizeable percentages of CD15+ and CD66b+ cells were MPO-. CONCLUSIONS High density MPO+ cell infiltration is a novel independent favorable prognostic factor in CRC

Immunobiology of IL-17A in human colorectal cancer

During the last decade tumor infiltration by immune cells has been recognized as a key factor determining clinical outcome. Whereas the presence within tumor tissues of defined lymphocytic populations, including cytotoxic CD8+ T cells and IFN-gamma-producing T-helper 1 cells has been univocally recognized to predict favorable prognosis, the clinical relevance and the pathophysiological role of IL-17-producing cells remain unclear. In some tumor types, including ovarian, prostate and lung cancer, a positive association between tumor infiltration by IL-17+ cells and prolonged patient survival has been reported. In contrast, in colorectal cancer (CRC), IL-17 expression has been shown to predict unfavorable clinical outcome and to weaken the beneficial effect of tumor infiltration by CD8+ T cells. However, restricted numbers of patients were analyzed and no functional data concerning the IL-17 source and the potential mechanisms underlying its negative effect were provided by these studies. In the enclosed work we have investigated prognostic significance, phenotype, and functional features of tumor-infiltrating IL-17-producing cells in human CRC. Upon analysis of a tissue micro-array (TMA) including 1400 cases of primary CRC, we found that tumor infiltration by IL-17+ cells was significantly associated with lower T (tumor border) and N (lymph nodes involvement) stage, but in contrast to previous findings, did not per se impact on overall patients survival. Interestingly, numbers of IL-17+ cells strongly correlated with those of CD8+ and CD16+ myeloperoxidase (MPO)+ neutrophils, which were predictive of better clinical outcome in the same patient cohort. Phenotypic analysis revealed that the majority of tumor infiltrating IL-17+ cells consisted of polyfunctional T helper 17 (Th17), producing, in addition to IL-17 a spectrum of pro-inflammatory cytokines and chemokines such as TNF-alpha??IL-21, IL-22, and GM-CSF, and, IFN-gamma? and IL-8. Interestingly, tumor-derived Th17 cells induced IL-8-dependent neutrophil migration and enhanced MPO release. Furthermore, tumor-derived Th17 cells favored the indirect recruitment of CD8+ T cells, by triggering chemokine release from tumor-associated endothelial cells. More surprisingly, CD8+ T cells were also directly recruited by Th17 cells in a CCL20 dependent manner. Importantly, the direct effect of Th17 proved sufficient to drive CD8+ T cells into an engineered CRC tissue-like structure. Our data suggest that CRC infiltrating Th17 cells can favor the recruitment of clinical relevant effector cells into the tumor site, therefore contributing to a more favorable clinical outcome. Altogether our findings unravel a positive role possibly played by tumor infiltrating polyfunctional Th17 cells in CRC and underline their pleiotropic effects beyond IL-17 production

Improving Cross-Program Collaboration with the Australian Red Cross Victorian Emergency Services

The Victorian Red Cross delivers services across four program areas: Emergency Services, Migrant Support, Community Program, and the Community Mobilisation team. The Victorian Red Cross is seeking to strengthen coordination across programs to better support the people they aid–this project aims to complement those efforts. To accomplish this, the team: developed a framework for analyzing coordination, applied the framework to describe Red Cross’ current system, and utilized the framework to make recommendations to address opportunities that emerge in the data

FinTech Project B23 - Enhancing Investment Management - Data Science

In the field of alternative investment management, big data presents challenges and opportunities, necessitating a shift in how firms approach information management. Our team collaborated with an alternative investment firm to reduce the amount of time analysts review potential and historical investment deal data. To achieve this goal, we executed a two-part project: the first part involved constructing a Power BI dashboard of key metrics for their Residential Mortgage-Backed Securities desk using Azure Data Lake, Python, Power BI, and PostgreSQL; the second part utilized the company’s proprietary Chat-GPT system and prompt engineering to develop a deal appraisal model using Python, Databricks, Azure Cognitive Search/AI Document Intelligence, and Amazon Textract

Restauro della cappellina Strozzi in palazzo Pazzi-Quaratesi a Firenze. Analisi dello stato di conservazione dei dipinti e studio delle metodologie e dei materiali impiegati nei precedenti interventi di restauro.

Le decorazioni della piccola cappella risalgono all'inizio del Seicento. Sebbene ora siano decorati solo la volta a botte e la cornice, alcuni piccoli frammenti di colore rinvenuti fanno pensare che un tempo fossero dipinte anche le pareti. Le decorazioni sono state oggetto di restauri, alcuni dei quali piuttosto recenti: in quella sede la pulitura è stata probabilmente troppo aggressiva e ha contribuito sia alla perdita di saturazione dei dipinti e di coesione delle dorature sia alla rimozione delle parti più delicate eseguite “a secco”. Successivamente, per sopperire all'aspetto deteriorato, sono stati eseguiti significativi interventi di ritocco pittorico e ridipintura sulle zone abrase, in particolare sulle fasce decorative della volta e sulla cornice. Anche quest'ultima è stata oggetto di ripetute operazioni di imbiancatura e scopritura: in alcune parti sono infatti presenti significativi segni di azione meccanica e alcuni residui di scialbo. Partendo da questa condizione conservativa, l'intervento di restauro è stato finalizzato al recupero di idonei parametri conservativi ed estetic

Ectosomes released by platelets induce differentiation of CD4+T cells into T regulatory cells

Accumulating evidence suggests an immune-modulatory role for platelets (PLT) and PLT-derived microvesicles. In particular, ectosomes, i.e. vesicles budding from PLT surface, have been shown to exert immunosuppressive activities on phagocytes. Here we investigated the effects mediated by PLT-derived ectosomes (PLT-Ecto) on CD4+ T cells. Exposure of activated CD4+ T cells to PLT-Ecto decreased their release of IFNgamma, TNFalpha and IL-6, and increased the production of TGF-beta1. Concomitantly, PLT-Ecto-exposed CD4+ T cells displayed increased frequencies of CD25high Foxp3+ cells. These phenomena were dose-dependent and PLT-Ecto specific, since they were not observed in the presence of polymorphonuclear- and erythrocyte-derived ectosomes. Analysis of specific T cell subsets revealed that PLT-Ecto induced differentiation of naive T cells into Foxp3+ cells, but had no effect on pre-differentiated Foxp3+ regulatory T cells (Tregs). Importantly, PLT-Ecto-induced Foxp3+ cells were as effective as peripheral blood Tregs in suppressing CD8+ T cell proliferation. PLT-Ecto-mediated effects were partly dependent on PLT-derived TGF-beta1, as they were to some extent inhibited by PLT-Ecto pretreatment with TGF-beta1-neutralising antibodies. Interestingly, ectosome-derived TGF-beta1 levels correlated with Foxp3+ T cell frequencies in blood of healthy donors. In conclusion, PLT-Ecto induce differentiation of CD4+ T cells towards functional Tregs. This may represent a mechanism by which PLT-Ecto enhance peripheral tolerance

Ectosomes released by platelets induce differentiation of CD4+ T cells into T regulatory cells

Accumulating evidence suggests an immune-modulatory role for platelets (PLT) and PLT-derived microvesicles. In particular, ectosomes, i.e. vesicles budding from PLT surface, have been shown to exert immunosuppressive activities on phagocytes. Here we investigated the effects mediated by PLT-derived ectosomes (PLT-Ecto) on CD4+ T cells. Exposure of activated CD4+ T cells to PLT-Ecto decreased their release of IFNgamma, TNFalpha and IL-6, and increased the production of TGF-beta1. Concomitantly, PLT-Ecto-exposed CD4+ T cells displayed increased frequencies of CD25high Foxp3+ cells. These phenomena were dose-dependent and PLT-Ecto specific, since they were not observed in the presence of polymorphonuclear- and erythrocyte-derived ectosomes. Analysis of specific T cell subsets revealed that PLT-Ecto induced differentiation of naive T cells into Foxp3+ cells, but had no effect on pre-differentiated Foxp3+ regulatory T cells (Tregs). Importantly, PLT-Ecto-induced Foxp3+ cells were as effective as peripheral blood Tregs in suppressing CD8+ T cell proliferation. PLT-Ecto-mediated effects were partly dependent on PLT-derived TGF-beta1, as they were to some extent inhibited by PLT-Ecto pretreatment with TGF-beta1-neutralising antibodies. Interestingly, ectosome-derived TGF-beta1 levels correlated with Foxp3+ T cell frequencies in blood of healthy donors. In conclusion, PLT-Ecto induce differentiation of CD4+ T cells towards functional Tregs. This may represent a mechanism by which PLT-Ecto enhance peripheral tolerance