Interaction between Human NK Cells and Bone Marrow Stromal Cells Induces NK Cell Triggering: Role of NKp30 and NKG2D Receptors

Abstract In this study we have analyzed the interaction between in vitro cultured bone marrow stromal cells (BMSC) and NK cells. Ex vivo-isolated NK cells neoexpressed the activation Ag CD69 and released IFN-γ and TNF-α upon binding with BMSC. Production of these proinflammatory cytokines was dependent on ligation of ICAM1 expressed on BMSC and its receptor LFA1 on NK cells. Furthermore, the NKp30, among natural cytotoxicity receptors, appeared to be primarily involved in triggering NK cells upon interaction with BMSC. Unexpectedly, autologous IL-2-activated NK cells killed BMSC. Again, LFA1/ICAM1 interaction plays a key role in NK/BMSC interaction; this interaction is followed by a strong intracellular calcium increase in NK cells. More importantly, NKG2D/MHC-I-related stress-inducible molecule A and/or NKG2D/UL-16 binding protein 3 engagement is responsible for the delivery of a lethal hit. It appears that HLA-I molecules do not protect BMSC from NK cell-mediated injury. Thus, NK cells, activated upon binding with BMSC, may regulate BMSC survival

Mesenchymal Stem Cell-Based Immunomodulation in Allogeneic Heterotopic Heart-Lung Transplantation

Mesenchymal stem cells are able to differentiate in various cell lineages and they have shown immunomodulatory properties in vitro, altering the cytokine secretion profile of T helper, T effector and dendritic cells and stimulating natural killer cells towards an anti-inflammatory and tolerant phenotype. In vivo they prolong skin allograft survival and may decrease graft-versus-host disease after hematopoietic stem cell transplants. In this work we studied the effects of mesenchymal stem cell treatment in an allogeneic heterotopic heart-lung transplant model. The following experimental groups were formed: A) Control B) Immunosuppressive therapy (Cyclosporine A) C) Mesenchymal stem-cell intravenous infusion D) Mesenchymal stem-cell infusion plus immunosuppressive treatment. The infusion of mesenchymal stem cells improved the mean graft survival up to 14.5±3.7 days with respect to the control group (3±0.6 days). Treatment with Cyclosporine A plus mesenchymal stem cells (group D) produced a mean survival time of 18.25±4.9 days, and was not significantly different to the results for group B (21.75±3.5 days). Furthermore, in the immunosuppressive treatment and the mesenchymal stem cell treatment, histological analysis revealed a reduction in the grade of rejection in heart and lung grafts. This decrease was most significant in group D. In conclusion, mesenchymal stem cells alone or in combination with Cyclosporine A were able to prolong graft survival time. These data suggest that, in vivo, mesenchymal stem cells retain their ability, already shown in vitro, to suppress lymphocyte activation and proliferation

Studio pilota sul ruolo dell\u2019allattamento e dei fattori riproduttivi nel rischio dei carcinomi mammari Luminali nelle donne in premenopausa

Il ruolo dei fattori riproduttivi nel rischio di sviluppare un carcinoma alla mammella \ue8 ancora controverso e poco si sa su come questi influiscano sul rischio dei differenti sottotipi molecolari . La maggior parte degli studi pubblicati riguardano coorti di donne in pre e postmenopausa o solo donne in postmenopausa. Scopo del nostro studio \ue8 stato analizzare il ruolo dell\u2019allattamento e dei fattori riproduttivi nei tumori Luminali, il 75% di tutti i carcinomi della mammella, in un campione di donne giovani residenti nella provincia di Trieste, situata in Friuli-Venezia Giulia, una tra le regioni con i pi\uf9 alti tassi d\u2019incidenza di carcinoma mammario

The clinical effectiveness of an integrated multidisciplinary evidence-based program to prevent intraoperative pressure injuries in high-risk children undergoing long-duration surgical procedures: a quality improvement study

The prevention of hospital-acquired pressure injuries (HAPIs) in children undergoing long-duration surgical procedures is of critical importance due to the potential for catastrophic sequelae of these generally preventable injuries for the child and their family. Long-duration surgical procedures in children have the potential to result in high rates of HAPI due to physiological factors and the difficulty or impossibility of repositioning these patients intraoperatively. We developed and implemented a multi-modal, multi-disciplinary translational HAPI prevention quality improvement program at a large European Paediatric University Teaching Hospital. The intervention comprised the establishment of wound prevention teams, modified HAPI risk assessment tools, specific education, and the use of prophylactic dressings and fluidized positioners during long-duration surgical procedures. As part of the evaluation of the effectiveness of the program in reducing intraoperative HAPI, we conducted a prospective cohort study of 200 children undergoing long-duration surgical procedures and compared their outcomes with a matched historical cohort of 200 children who had undergone similar surgery the previous year. The findings demonstrated a reduction in HAPI in the intervention cohort of 80% (p < 0.01) compared to the comparator group when controlling for age, pathology, comorbidity, and surgical duration. We believe that the findings demonstrate that it is possible to significantly decrease HAPI incidence in these highly vulnerable children by using an evidence-based, multi-modal, multidisciplinary HAPI prevention strategy

Gestione delle pazienti con tumore fillode della mammella: esperienza triestina nel periodo 2006-2014

La diagnosi e la gestione dei tumori fillodi della mammella \ue8 complessa a causa del basso tasso di incidenza e dell\u2019imprevedibilit\ue0 del comportamento di questo tipo di neoplasie (meno dell\u20191% tra tutti i tumori della mammella [1]). L\u2019obiettivo di questo studio \ue8 analizzare i casi di tumori filloidi diagnosticati a Trieste nel periodo 2006-2014 al fine di contestualizzare il comportamento particolarmente aggressivo di un tumore fillode maligno insorto in una paziente con pregressi fillodi benigni

Therapeutic strategy for ductal carcinoma in situ patients according to Van Nuys Prognostic Index

Aim: Evaluation of therapeutic strategy for Ductal Carcinoma In Situ (DCIS) patients at our Brest Cancer Centre and analysis of our pattern of treatment with respect to Van Nuys Prognostic Index (VNPI) cathegories. Matherial of Study: our study population is the result of a selction of 85 DCIS patients classified according to VNPI risk of local recurrence (LR). A comparison was made between treatment affectiverly performed and therapy suggested by VNPI protocols. RESULTS: Out of 53 DCIS women (62%) at low local recurrence risk, 5 patients underwent Breast Conserving Surgery (BCS) alone, 7 were treated with mastectomy and 41 underwent BCS followed by radiotherapy (RT). Out of 31 patients (37%) belonging to VNPI intermediate risk group, 25 cases recived BCS+RT and 6 cases received mastectomy. Only one patient (1%) belonged to VNPI high risk group and underwent mastectomy. DISCUSSION: Only 31 patients (36,5%) had their definitive treatment according to recommended VNPI criteria, but none of the other 54 cases (63,5%) was undertreated. Performing mastectomy instead of BCS or adding adjuvant radiotherapy at BCS alone were not considered overtreatment because the therapeutic strategy was the result of a multidisciplinary discussion. CONCLUSION: As DCIS is a heterogeneus desease the one-size-fits-all approach to treatment seems inappropriate. The VNPI was developed in order to help treatment choices, but therapeutic strategies can\u2019t be based only on local recurrence risk and need a multidisciplinary approach

SHOTGUN PROTEOMICS OF HUMAN PANCREATIC ISLETS: EFFECT OF CYTOKINES EXPOSURE

Background: The effect of cytokines on β-cell function and survival was studied in β-cells as INS1-E and a different role was suggested for INF-γ and IL-1 which act on activation of cellular defense mechanism and cell functions, respectively. In addition, pro-inflammatory cytokines also showed to induce β-cell apoptosis and endoplasmic reticulum stress. In order to investigate the effect of cytokines on human islets, the complete proteome before and after exposure to cytokines was analyzed using label free shot-gun proteomics. Methods: Aliquots of 40 µg of human islets protein extracts (n=3), with and without cytokines treatment, were loaded onto 12% acrylamide resolving gel. After separation, gel pieces (13 pieces for lane) were excised from the gel and the proteins were identified by Shotgun methodology after an in-gel trypsin digestion. Mass spectrometry data were acquired according to the novel label free quantitation workflow developed by Bruker Daltonik. Results: Around 3000 proteins were identified and out of 307 differentially expressed proteins, 184 resulted increased (among these chemokines, oxidative stress related proteins and immunoproteasome proteins) and 123 reduced (i.e cathepsines, antioxidant proteins, Krebs enzymes) after treatment with cytokines. Ingenuity pathways analysis highlighted the activation of upstream regulators such as STAT1 and 2, NFKb, JAK1, and inhibition of MAPK1, atypical chemokine receptor 2, transcription intermediary factor 1-alpha and small ubiquitin-related modifier 3. Finally, the treatment with cytokines induced a significant decrease (-28±10%) of insulin secretion with respect to control, together with an increase of apoptotic beta cells and a reduction of volume density of grain of insulin. Conclusions: Overall our results show how the detrimental effects of cytokines treatment in human pancreatic islets could be associated to proteome changes. Further studies are necessary to clarify the correlation between these deregulation and type 1 diabetes features. Keywords: cytokines, human pancreatic islets, type-1 diabetes

Shotgun proteomics of human pancreatic islets: effect of cytokines exposure

The effect of cytokines on β-cell function and survival was studied in β-cells as INS1-E and a different role was suggested for INF-γ and IL-1 (Rondas et al, J Proteome Res, 2013). In order to investigate the effect of cytokines on human islets, the complete proteome before and after exposure to cytokines was analyzed using label free shot-gun proteomics. Aliquots of 40 µg of human islets protein extracts, with and without cytokines treatment, were loaded onto 12% acrylamide resolving gel. After separation, gel pieces were excised from the gel and the proteins were identified by shot-gun after in-gel trypsin digestion. Around 3000 proteins were identified and out of 245 differentially expressed proteins, 158 resulted increased and 87 reduced after treatment with cytokines. Overall these proteins appertain mainly to metabolism, defense, repair and immunoresponse, protein synthesis and degradation. By IPA analysis the network and the involved canonical pathways were suggested. Moreover, 154 predicted upstream regulators (118 activated and 36 inhibited) were prompted. In addition to known regulator factors such as STAT1, NFKb, IRF1, MAPK1, new ones as HMGB1, GPSA and SIRT1 were found. Finally, the cytokines induced a significant decrease of insulin secretion respect to control, together with an increase of apoptotic beta cells and a reduction of volume density of grain of insulin. Overall our results show how the detrimental effects of cytokines treatment in human pancreatic islets could be associated to proteome changes. Further studies are necessary to clarify the correlation between these deregulation and type 1 diabetes features