Evaluation of the efficacy of intrahepatic autologous stem cells (SC) CD133+ infusion in patients with cirrhosis and advanced hepatic insufficiency

Numerose evidenze sperimentali hanno dimostrato il contributo delle cellule staminali (SC) di derivazione midollare nei processi di rigenerazione epatica dopo danno tissutale. E’ cresciuto pertanto l’interesse sul loro potenziale impiego in pazienti con cirrosi. Questo studio si proponeva di valutare la fattibilità e la sicurezza della reinfusione intraepatica di cellule staminali midollari autologhe CD133+ in 12 pazienti con insufficienza epatica terminale. Previa mobilizzazione nel sangue periferico mediante somministrazione di granulocyte-colony stimulating factor (G-CSF) alla dose di 7,5 mcg/Kg/b.i.d. e raccolta per leucoaferesi (solo se la concentrazione di CD133 + SC era > 8/μL), le cellule CD133+ altamente purificate sono state reinfuse in arteria epatica a partire da 5x104/Kg fino a 1x106/kg. Nei tre giorni successivi è stato somministrato G-CSF per favorire l’espansione e l’attecchimento delle cellule. Durante la fase della mobilizzazione e quella della reinfusione sono stati eseguiti saggi biologici quali: caratterizzazione fenotipica delle SC circolanti, saggi clonogenici, valutazione della concentrazione sierica del Hepatocyte Growth Factor (HGF), Stromal-Derived Factor-1 (SDF-1) ed il Vascular-Endotelial Growth Factor (VEGF) e caratterizzazione fenotipica delle CD133+SC purificate. Fino ad oggi sono stati reinfusi 12 pazienti. Questi dati preliminari suggeriscono che è possibile mobilizzare e reinfondere un numero considerevole di SC autologhe CD133+ altamente purificate in pazienti con ESLD . Gli studi biologici mostrano che: il numero di progenitori ematopoietici ed endoteliali circolanti è aumentato dopo il trattamento con G–CSF; le SCs CD133+ altamente purificato esprimono marcatori emopoietici ed endoteliali; la concentrazione sierica di HGF, SDF-1, VEGF e la capacità clonogenica di progenitori emopoietici sono aumentati durante la mobilitazione e nelle fasi di reinfusione; il potenziale clonogenico dei progenitori endoteliali mostra espressione variabile.Bone marrow (BM) stem cells (SCs) have been shown to contribute to liver cell populations and this has sparked interest in the field of autologous SCs infusion as a possible treatment for cirrhosis. The aim of this study was to evaluate the feasibility and safety of intrahepatic reinfusion of increasing numbers of autologous BM-derived CD133+ SCs into hepatic artery of 12 patients with end-stage liver disease (ESLD). For this purpose, granulocyte-colony-stimulating factor (G-CSF) at 7.5 µg/Kg/b.i.d. was administered subcutaneously (sc) from day 1 until completing the peripheral blood stem cells (PBSCs) collection. PBSCs collection started on day 5 only if the CD133+SCs concentration was >8/µL. CliniMacs device was used for the positive selection of CD133+SCs from PB of mobilized standard-volume leukapheresis. After SCs mobilization, highly purified autologous G-CSF-mobilized CD133+SCs were reinfused through hepatic artery. CD133+CSs were administered according to body weight starting from 5x104/Kg and increased every 3 patients up to 1x106/Kg. G-CSF at 5µg/Kg/day was administered sc for 3 days after reinfusion of SCs for their expansion and to induce a selective proliferative advantage in vivo. Biological assays (circulating SCs phenotype, clonogenic assays, serum concentration of hepatocyte growth factor [HGF], stromal-derived factor-1 [SDF-1] and vascular-endotelial growth factor [VEGF]) were done during the mobilization and reinfusion phases together with the phenotypic characterization of the isolated CD133+SCs. Up to date, 12 patients have been reinfused. These preliminary data suggest that it is feasible to mobilize and reinfuse a substantial number of highly purified autologous CD133+ SCs in patients with ESLD. Biological studies show that: circulating hematopoietic and endothelial progenitors are increased after G-CSF treatment; highly purified CD133+CSs express hematopoietic and endothelial markers; serum concentration of HGF, SDF-1, VEGF and clonogenic capability of hematopoietic progenitors are increased during the mobilization and reinfusion phases; clonogenic potential of endothelial progenitors shows variable expression

Stem cells for end stage liver disease: How far have we got?

End stage liver disease (ESLD) is a health problem worldwide. Liver transplantation is currently the only effective therapy, but its many drawbacks include a shortage of donors, operative damage, risk of rejection and in some cases recidivism of the pre-transplant disease. These factors account for the recent growing interest in regenerative medicine. Experiments have sought to identify an optimal source of stem cells, sufficient to generate large amounts of hepatocytes to be used in bioartificial livers or injected in vivo to repair the diseased organ. This update aims to give non-stem cell specialists an overview of the results obtained to date in this fascinating field of biomedical research

Biotransformation of Waste Bile Acids: A New Possible Sustainable Approach to Anti-Fungal Molecules for Crop Plant Bioprotection?

Phytopathogenic fungi are among the main causes of productivity losses in agriculture. To date, synthetic chemical pesticides, such as hydroxyanilides, anilinopyrimidines and azole derivatives, represent the main treatment tools for crop plant defence. However, the large and uncontrolled use of these substances has evidenced several side effects, namely the resistance to treatments, environmental damage and human health risks. The general trend is to replace chemicals with natural molecules in order to reduce these side effects. Moreover, the valorisation of agri-food industry by-products through biotransformation processes represents a sustainable alternative to chemical synthesis in several sectors. This research is aimed at comparing the anti-phytopathogenic activity of waste bovine and porcine bile with secosteroids obtained by biotransformation of bile acids with Rhodococcus strains. The ultimate goal is to apply these natural products on food crops affected by phytopathogenic fungi

Serum hepatitis B core-related antigen is an effective tool to categorize patients with HBeAg-negative chronic hepatitis B

The discrimination between active chronic hepatitis B (CHB) and the clinically quiescent infection (CIB) is not always easy, as a significant portion of patients falls in a “grey” zone. Hepatitis B core-related antigen (HBcrAg) is a now quantifiable serological marker with potential applications in diagnosis and therapy monitoring. The aim of the present study was to evaluate the HBcrAg serum levels in HBeAg-negative HBV infection, and its ability in identifying the clinical profile, in comparison with HBsAg serum levels. HBcrAg was retrospectively assessed on serum samples from a population of treatment-naive HBeAg-negative patients by ChemiLuminescent Enzyme Immunoassay (CLEIA). HBsAg and HBV-DNA data were collected. Serological data were associated to clinical profile, defined in the subsequent follow-up of at least 1 year. In the overall population of 160 HBeAg-negative patients, HBcrAg results weakly correlated with qHBsAg levels (Spearman r = 0.471, P < 0.0001) and correlated closely with HBV-DNA (Spearman r = 0.746, P < 0.0001). HBcrAg levels were significantly higher in 85 CHB patients relative to 75 CIB carriers. A value of 2.5 logU/mL produced the optimal cut-off to identify CIB patients, with diagnostic accuracy comparable to HBsAg levels. In long-term clinical evaluation, a single measurement of HBcrAg at the established cut-off was optimally consistent with clinical outcome. Conversely, the HBsAg cut-off performed well in the true quiescent phase and less in more difficult-to-categorize patients. In conclusion, single-point use of HBcrAg serum levels provides an accurate identification of CIB and represents a useful tool for patient classification

In vitro effect of thymosin-alpha1 and interferon-alpha on Th1 and Th2 cytokine synthesis in patients with eAg-negative chronic hepatitis B

none11mixedLoggi E; Gramenzi A; Margotti M; Cursaro C; Galli S; Vitale G; Grandini E; Scuteri A; Vukotic R; Andreone P; Bernardi M.Loggi E; Gramenzi A; Margotti M; Cursaro C; Galli S; Vitale G; Grandini E; Scuteri A; Vukotic R; Andreone P; Bernardi M

Immunological modifications during treatment with thymosin alpha 1 plus antiviral therapy in chronic hepatitis C

The current standard therapy for the treatment of chronic hepatitis C virus (HCV) is the combination of peginterferon and ribavirin, although many patients fail to clear the virus and their retreatment options are still unsatisfactory. Thymosin alpha1 (Talpha1) is an immunomodulating agent that has been proposed as complementary therapy for chronic HCV, especially in the setting of difficult-to-treat patients. The aim of this study was to evaluate, in patients nonresponsive to previous Peg-based therapy, the effect of standard antiviral therapy with or without Talpha1 on peripheral lymphocyte subsets. Twenty-four patients, 12 receiving Talpha1 and 12 standard therapy, were enrolled. Peripheral subpopulations were analyzed by flow cytometry. Although the addition of Talpha1 did not seem to significantly modify the T-lymphocyte subpopulations, as comparable behaviors were observed in the CD4 and CD8 longitudinal evaluation, Talpha1 produced an earlier increase of natural killer cells. An accurate selection of HCV patients who can benefit from immunomodulation is neede

Deregulation of microRNA expression in peripheral blood mononuclear cells from patients with HCV-related malignancies

Background and aim: Hepatocellular carcinoma is one of the major causes of death due to cancer worldwide, and its association with hepatitis C virus infection has been definitively established. Hepatitis C virus is also involved in the pathogenesis of non-Hodgkin\u2019s lymphoma. This is the only virus infecting humans that is able to induce two different malignancies. We analyzed the expression levels of a panel of microRNA in peripheral blood mononuclear cells of patients with hepatitis C virus-related malignancies in order to find a disease-associated deregulation and identify specific biomarkers. Methods: We tested peripheral blood mononuclear cells isolated from patients with hepatocellular carcinoma, non-Hodgkin\u2019s lymphoma, hepatitis C virus without malignancies and healthy subjects for a panel of microRNA selected on the basis of previous studies. MicroRNA expression was evaluated by real-time PCR. Results: Our results showed an upregulation of miRNA-21 and downregulation of miRNA-26b in hepatocellular carcinoma and non-Hodgkin\u2019s lymphoma patients compared to controls (p\ua0<\ua00.001). Deregulation of miRNA-16 and miRNA-155 was limited to lymphoma patients. Conclusions: This study shows that some microRNAs are differently expressed in peripheral blood mononuclear cells from hepatitis C virus patients who develop hepatocellular carcinoma or lymphoma, while others share a common behavior. Thus, analysis of the expression of microRNAs could be a noninvasive marker of hepatitis C virus-related carcinogenesis. This analysis could be a suitable tool for identifying the existence of a malignancy and also discriminating between these two hepatitis C virus-related cancers