New therapies, markers and therapeutic targets in HCV chronic infection, and HCV extrahepatic manifestations

More than 180 millions of subjects in the world are infected by Hepatitis C Virus (HCV), and about 20% of them with HCV chronic infection progress to cirrhosis. Furthermore, numerous HCV extrahepatic manifestations have been reported in up to 74% of patients, as mixed cryoglobulinemia, lymphomas, rheumatic disorders, autoimmune thyroiditis, hypothyroidism, papillary thyroid cancer, and type 2 diabetes. Advances in understanding the HCV life cycle, and the inflammatory processes (involving a complex network of cytokines and chemokines) associated with HCV chronic infection, have led to substantial advancements in therapy. The combination of ribavirin and PEGylated interferon-gamma was the standard of therapy for HCV chronically infected patients in the last decades. However, interferon has limited effectiveness and is associated with severe adverse effects. Recently, direct-acting antivirals (DAAs) that act as inhibitors of N5SA, or polymerase, or protease have been shown to result in shorter duration of therapy, better efficacy and tolerance, with respect to ribavirin and PEGylated interferon-gamma. Circulating CXCL10 levels, and the interleukin(IL)-28B gene polymorphisms, are associated with the success of the therapy both with DAAs or ribavirin and PEGylated interferon-alpha. New DAAs targeting the HCV at various molecular levels have been developed to eradicate HCV. Moving to interferon-free therapies should offer new treatments for resistant HCV genotypes, and for ineligible patients or patients failing to respond to prior therapies. Many efforts have been made to understand the factors that are involved with clearance of HCV to personalize the therapy for each patient, with the aim to reduce side effects, increasing the sustained virologic response rate, and to prevent the progression of the disease

Integration of the viral genome into the host cell genome: a double-edged sword

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Varicella-zoster virus infection: natural history, clinical manifestations, immunity and current and future vaccination strategies

Varicella-zoster virus (VZV) is the etiologic agent of varicella (chicken pox), a childhood exanthematic disease that develops as a result of primary infection, and zoster (shingles), caused by reactivation of the virus persisting in a latent form in the dorsal sensory ganglia. Although varicella is generally a mild self-limiting illness, in immunocompromised subjects and adults it can have a serious clinical course that can lead to permanent damage of the central nervous system. In these and in most zoster cases, treatment with anti-herpetic drugs and/or immunotherapy is necessary. Because it is highly contagious, varicella is one of the most common exanthematic diseases. It is preventable by vaccination with an attenuated vaccine administered around the first year of age, and with a boost vaccination in school age. This article briefly describes the natural history and pathophysiology of VZV infection and its current epidemiology and provides an overview of current and future vaccine options to protect against varicella and/or zoster

Evaluation of T cell immunity against human cytomegalovirus: Impact on patient management and risk assessment of vertical transmission

Cytomegalovirus (CMV) is one of the most common infectious agents, infecting the general population at an early age without causing morbidity most of the time. However, on particular occasions, it may represent a serious risk, as active infection is associated with rejection and disease after solid organ transplantation or fetal transmission during pregnancy. Several methods for CMV diagnosis are available on the market, but because infection is so common, careful selection is needed to discriminate primary infection from reactivation. This review focuses on methods based on CMV-specific T cell reactivity to help monitor the consequences of CMV infection/reactivation in specific categories of patients. This review makes an attempt at discussing the pros and cons of the methods available

How Current Direct-Acting Antiviral and Novel Cell Culture Systems for HCV are Shaping Therapy and Molecular Diagnosis of Chronic HCV Infection

We have entered a new era of hepatitis C virus (HCV) therapy in which elimination of infection and disease is a real possibility. HCV cell culture models were instrumental for identification of therapeutic targets, testing candidate drugs, and profiling of therapeutic strategies. Here we describe current and novel methods of cell culture systems for HCV that are allowing investigation of HCV life cycle and virus-host interaction required for replication and propagation. The development of protocols to grow infectious virus in culture and generate hepatocyte cell lines from specific individuals hold great promise to investigate the mechanisms exploited by the virus to spread the infection and the host factors critical for HCV replication and propagation, or resistance to infection. Since host factors are presumably conserved and equally interacting with different HCV isolates and genotypes, the development of drugs targeting host factors essential for virus replication holds great promises in further increasing treatment efficacy. Refocusing of therapeutic goals also impacted in vitro diagnosis. The primary goal of anti-HCV therapy is to achieve a sustained virologic response (SVR) defined as " undetectable" HCV RNA genome in the serum or plasma at 12 to 24 weeks following the end of treatment. Use of direct antiviral agents has substantially changed the threshold of the viral load used to define SVR and led to a reassessment, as discussed herein, of result interpretation and requirements of clinically-approved, quantitative molecular assays

Tweaking Mesenchymal Stem/Progenitor Cell Immunomodulatory Properties with Viral Vectors Delivering Cytokines

Mesenchymal Stem Cells (MSCs) can be found in various body sites. Their main role is to differentiate into cartilage, bone, muscle, and fat cells to allow tissue maintenance and repair. During inflammation, MSCs exhibit important immunomodulatory properties that are not constitutive, but require activation, upon which they may exert immunosuppressive functions. MSCs are defined as "sensors of inflammation" since they modulate their ability of interfering with the immune system both in vitro and in vivo upon interaction with different factors. MSCs may influence immune responses through different mechanisms, such as direct cell-to-cell contact, release of soluble factors, and through the induction of anergy and apoptosis. Human MSCs are defined as plastic-adherent cells expressing specific surface molecules. Lack of MHC class II antigens makes them appealing as allogeneic tools for the therapy of both autoimmune diseases and cancer. MSC therapeutic potential could be highly enhanced by the expression of exogenous cytokines provided by transduction with viral vectors. In this review, we attempt to summarize the results of a great number of in vitro and in vivo studies aimed at improving the ability of MSCs as immunomodulators in the therapy of autoimmune, degenerative diseases and cancer. We will also compare results obtained with different vectors to deliver heterologous genes to these cells

A novel method for producing target cells and assessing cytotoxic T lymphocyte activity in outbred hosts

<p>Abstract</p> <p>Background</p> <p>Cytotoxic T lymphocytes play a crucial role in the immunological control of microbial infections and in the design of vaccines and immunotherapies. Measurement of cytotoxic T lymphocyte activity requires that the test antigen is presented by target cells having the same or compatible class I major hystocompatibility complex antigens as the effector cells. Conventional assays use target cells labeled with ⁵¹chromium and infer cytotoxic T lymphocyte activity by measuring the isotope released by the target cells lysed following incubation with antigen-specific cytotoxic T lymphocytes. This assay is sensitive but needs manipulation and disposal of hazardous radioactive reagents and provides a bulk estimate of the reporter released, which may be influenced by spontaneous release of the label and other poorly controllable variables. Here we describe a novel method for producing target in outbred hosts and assessing cytotoxic T lymphocyte activity by flow cytometry.</p> <p>Results</p> <p>The method consists of culturing skin fibroblasts, immortalizing them with a replication defective clone of simian virus 40, and finally transducing them with a bicistronic vector encoding the target antigen and the reporter green fluorescent protein. When used in a flow cytometry-based assay, the target cells obtained with this method proved valuable for assessing the viral envelope protein specific cytotoxic T lymphocyte activity in domestic cats acutely or chronically infected with feline immunodeficiency virus, a lentivirus similar to human immunodeficiency virus and used as animal model for AIDS studies.</p> <p>Conclusion</p> <p>Given the versatility of the bicistronic vector used, its ability to deliver multiple and large transgenes in target cells, and its extremely wide cell specificity when pseudotyped with the vesicular stomatitis virus envelope protein, the method is potentially exploitable in many animal species.</p

Small Anellovirus in Hepatitis C Patients and Healthy Controls

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