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Interferon (IFN) is a multifaceted immunomodulator that is effective against many diseases, including chronic hepatitis B and chronic hepatitis C infection. IFN defends against viral infection, but may also cause various side effects, such as ulcerative colitis (UC). Herein, we present a case of UC triggered by pegylated interferon alpha-2b (PEG-IFN-α-2b) therapy in a patient with concurrent chronic hepatitis B (CHB) infection. The diagnosis was based on typical clinical symptoms, colonoscopy findings, colonic mucosal biopsy, and histopathology. Accordingly, treatment with mesalazine was initiated without stopping PEG-IFN-α-2b. Fortunately, UC relieved gradually without compromising the effects of treatment. Simultaneously, we conducted a literature review of previously published case reports on the side effect of UC in patients with underlying chronic hepatitis. Various reactions have been reported, including induction, exacerbation, and no change. This is the first report of UC triggered by PEG-IFN-α-2b in a CHB patient. We recommend that physicians pay attention to the rare side effect of UC during administration of PEG-IFN-α-2b. Mesalazine can relieve UC with sustained use of PEG-IFN-α-2b.

Background Currently, there are no drugs that have been proven to be effective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Because of its broad antiviral activity, interferon (IFN) should be evaluated as a potential therapeutic agent for treatment of coronavirus disease 2019 (COVID-19), especially while COVID-19-specific therapies are still under development. Methods Confirmed COVID-19 patients hospitalized in the First Affiliated Hospital, School of Medicine, Zhejiang University in Hangzhou, China, from January 19 to February 19, 2020 were enrolled in a retrospective study. The patients were separated into an IFN group and a control group according to whether they received initial IFN-α2b inhalation treatment after admission. Propensity-score matching was used to balance the confounding factors. Results A total of 104 confirmed COVID-19 patients, 68 in the IFN group and 36 in the control group, were enrolled. Less hypertension (27.9% vs. 55.6%, P=0.006), dyspnea (8.8% vs. 25.0%, P=0.025), or diarrhea (4.4% vs. 19.4%, P=0.030) was observed in the IFN group. Lower levels of albumin and C-reactive protein and higher level of sodium were observed in the IFN group. Glucocorticoid dosage was lower in the IFN group (median, 40 vs. 80 mg/d, P=0.025). Compared to the control group, fewer patients in the IFN group were ventilated (13.2% vs. 33.3%, P=0.015) and admitted to intensive care unit (ICU) (16.2% vs. 44.4%, P=0.002). There were also fewer critical patients in the IFN group (7.4% vs. 25.0%, P=0.017) upon admission. Although complications during admission process were comparable between groups, the discharge rate (85.3% vs. 66.7%, P=0.027) was higher and the hospitalization time (16 vs. 21 d, P=0.015) was shorter in the IFN group. When other confounding factors were not considered, virus shedding time (10 vs. 13 d, P=0.014) was also shorter in the IFN group. However, when the influence of other factors was eliminated using propensity score matching, virus shedding time was not significantly shorter than that of the control group (12 vs. 15 d, P=0.206). Conclusions IFN-α2b spray inhalation did not shorten virus shedding time of SARS-CoV-2 in hospitalized patients.

Apolipoprotein L1 (APOL1) risk variants G1 and G2 are known to result in risk for kidney disease in patients of African ancestry. APOL1-associated nephropathy typically occurs in association with certain environmental factors or systemic diseases. As such, there has been increasing evidence of the role of interferon (IFN) pathways in the pathogenesis of APOL1-associated collapsing glomerulopathy in patients with human immunodeficiency virus (HIV) infection and systemic lupus erythematosus, 2 conditions that are associated with high IFN levels. Collapsing glomerulopathy has also been described in patients receiving exogenous IFN therapy administered for various medical conditions. We describe a patient with a genetic condition that results in an increased IFN state, stimulator of IFN genes (STING)-associated vasculopathy with onset in infancy (SAVI), who developed collapsing glomerulopathy during a flare of his disease. The patient was found to have APOL1 G1 and G2 risk variants. This case supports the role of IFN in inducing APOL1-associated collapsing glomerulopathy.

The IFITM restriction factors play a role in cancer cell progression through undefined mechanisms. We investigate new protein–protein interactions for IFITM1/3 in the context of cancer that would shed some light on how IFITM1/3 attenuate the expression of targeted proteins such as HLA-B. SBP-tagged IFITM1 protein was used to identify an association of IFITM1 protein with the SRSF1 splicing factor and transporter of mRNA to the ribosome. Using in situ proximity ligation assays, we confirmed a predominant cytosolic protein–protein association for SRSF1 and IFITM1/3. Accordingly, IFITM1/3 interacted with HLA-B mRNA in response to IFNγ stimulation using RNA–protein proximity ligation assays. In addition, RT-qPCR assays in IFITM1/IFITM3 null cells and wt-SiHa cells indicated that HLA-B gene expression at the mRNA level does not account for lowered HLA-B protein synthesis in response to IFNγ. Complementary, shotgun RNA sequencing did not show major transcript differences between IFITM1/IFITM3 null cells and wt-SiHa cells. Furthermore, ribosome profiling using sucrose gradient sedimentation identified a reduction in 80S ribosomal fraction an IFITM1/IFITM3 null cells compared to wild type. It was partially reverted by IFITM1/3 complementation. Our data link IFITM1/3 proteins to HLA-B mRNA and SRSF1 and, all together, our results begin to elucidate how IFITM1/3 catalyze the synthesis of target proteins. IFITMs are widely studied for their role in inhibiting viruses, and multiple studies have associated IFITMs with cancer progression. Our study has identified new proteins associated with IFITMs which support their role in mediating protein expression; a pivotal function that is highly relevant for viral infection and cancer progression. Our results suggest that IFITM1/3 affect the expression of targeted proteins; among them, we identified HLA-B. Changes in HLA-B expression could impact the presentation and recognition of oncogenic antigens on the cell surface by cytotoxic T cells and, ultimately, limit tumor cell eradication. In addition, the role of IFITMs in mediating protein abundance is relevant, as it has the potential for regulating the expression of viral and oncogenic proteins.

Foot-and-mouth disease (FMD) is a highly contagious vesicular disease of cloven-hoofed animals that severely constrains international trade of livestock and animal products. Currently, disease control measures include broad surveillance, enforcement of sanitary policy, and use of an inactivated vaccine. While use of these measures has contributed to eliminating foot-and-mouth disease virus (FMDV) from a vast area of the world, the disease remains endemic in three continents, and outbreaks occasionally appear in previously declared FMD-free zones, causing economic and social devastation. Among others, a very fast rate of viral replication and the need for 7 days to achieve vaccine-induced protection are the main limitations in controlling the disease. New fast-acting antiviral strategies targeted to boost the innate immunity of the host to block viral replication are needed. Here we review the knowledge on the multiple strategies FMDV has evolved to block the host innate immunity, with particularly focus on the past and current research toward the development of interferon (IFN)-based biotherapeutics in relevant livestock species.

© 2019 Nogales, Aydillo, Ávila-Pérez, Escalera, Chiem, Cadagan, DeDiego, Li, García-Sastre and Martínez-Sobrido., Influenza viruses are important pathogens that affect multiple animal species, including humans. There are four types of influenza viruses: A, B, C, and D (IAV, IBV, ICV, and IDV, respectively). IAV and IBV are currently circulating in humans and are responsible of seasonal epidemics (IAV and IBV) and occasional pandemics (IAV). ICV is known to cause mild infections in humans and pigs, while the recently identified IDV primarily affect cattle and pigs. Influenza non-structural protein 1 (NS1) is a multifunctional protein encoded by the NS segment in all influenza types. The main function of NS1 is to counteract the host antiviral defense, including the production of interferon (IFN) and IFN-stimulated genes (ISGs), and therefore is considered an important viral pathogenic factor. Despite of homologous functions, the NS1 protein from the diverse influenza types share little amino acid sequence identity, suggesting possible differences in their mechanism(s) of action, interaction(s) with host factors, and contribution to viral replication and/or pathogenesis. In addition, although the NS1 protein of IAV, IBV and, to some extent ICV, have been previously studied, it is unclear if IDV NS1 has similar properties. Using an approach that allow us to express NS1 independently of the nuclear export protein from the viral NS segment, we have generated recombinant IAV expressing IAV, IBV, ICV, and IDV NS1 proteins. Although recombinant viruses expressing heterotypic (IBV, ICV, and IDV) NS1 proteins were able to replicate similarly in canine MDCK cells, their viral fitness was impaired in human A549 cells and they were highly attenuated in vivo. Our data suggest that despite the similarities to effectively counteract innate immune responses in vitro, the NS1 proteins of IBV, ICV, or IDV do not fully complement the functions of IAV NS1, resulting in deficient viral replication and pathogenesis in vivo., This research was partially funded by the New York Influenza Center of Excellence (NYICE), a member of the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Department of Health and Human Services, Centers of Excellence for Influenza Research and Surveillance (CEIRS) Contract No. HHSN272201400005C (NYICE); by the Department of Defense (DoD) Peer Reviewed Medical Research Program (PRMRP) grant W81XWH-18-1-0460 to LM-S; by the NIH R01AI141889 to FL; and by the Center for Research in Influenza Pathogenesis (CRIP, CEIRS Contract No. HHSN272201400008C to AG-S).

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) represents one of the most important viruses affecting the swine population with a huge impact on pig production in many countries. Moreover, the recent emergence of some highly pathogenic isolates (HP-PRRSV-1) has caused more severe economic losses. The effects of PRRSV on the immune system is poorly understood, indeed, the infection and persistence of the virus is related to a complex interaction with the immune system that determines a dysregulation on the mechanisms and the secretion of important cytokines of the host. The virus shows a restricted tropism for cells from the monocyte-macrophage-lineage, the main in vivo target cells is represented by porcine alveolar macrophages (PAM). The aim of the present thesis is to analysing the response of the macrophages population to the infection of different PRRSV isolates, both in vitro and in vivo. In the first in vitro study, we used as a model, monocytes derived macrophages (MDMs) pre-treated with different cytokines as IFNγ, IL-4 and IFNβ. Nine PRRSV-1 isolates were analyzed: two Italian strains, five Eastern European strains, and Lena and Lelystad as reference-strains for HP and low pathogenic PRRSV, respectively. The different strains were able to infect MDMs, with the best efficiency in unpolarized MØ and IL-4 treated MDMs, while the IFN-treatment determined an antiviral state, this evidence was most pronounced with IFNβ. In general, the high pathogenicity isolates infected a higher percentage of MDMs and replicated to higher titers. Regarding the cytokines measurement, IFNα and IL-10 were not detected in the supernatant of infected MDMs; moreover, the Italian PR40 strain was the only one that induced a significant release of TNF-α and IL1-β. The second study was performed by using as a model the polarized-porcine alveolar macrophages (PAMs) pre-treated with IFNγ, IL-4 and IFNβ, in order to compare two PRRSV-1.1 Italian strains, the normal pathogenic PR11 strain and the HP-PRRSV PR40. Interesting, we did not see difference between the two isolates, and any difference in the infectivity among Mock, IL-4 and IFNγ treatments, that evidence could found explanation in a re-polarization from the M1 to M2 phenotype after the infection. While, the infection of both viruses was completely blocked by IFNβ-treatment, highlighting the antiviral state induced by Interferon. Moreover, both strains did not induce the production of the cytokines tested: IFNα, IL-10 and TNF- α. The in vivo study consisted in two different experiments, the aim of the first one was to analyse the impact of two Italian PRRSV-1 subtype 1 strains (PR11 and PR40), with different in vivo pathogenicity, on the macrophages population of the thymus, as target cell for both PRRSV replication and response of the host immune system. The second experiment aimed at evaluating the effect of a heterologous vaccination on macrophages population of the thymus after a challenge infection with a HP-PRRSV. In order to define the macrophages population, immunohistochemistry for the N protein of the virus, TUNEL labelling and immunolabeling for CD163, CD172a, CD107a and BA4D5 was performed. The thymus of the animals infected with PRRSV strains, PR11 and PR40 dead at 10-14 dpi showed more severe lesions and the higher number of macrophages in all the compartments of the thymus gland compared with the others groups. This last result suggests a strong and early inflammatory process, without any difference between the normal and highly pathogenic strain. Riassunto Il virus della sindrome respiratoria e riproduttiva suina (PRRSV) è considerato uno dei più importanti virus del suino, con un grave impatto economico sulla produzione suinicola mondiale. Recentemente, sono emersi nuovi isolati del virus, caratterizzati da maggiore patogenicità e mortalità, e quindi definiti ceppi ad alta patogenicità (HP-PRRSV). Gli effetti dell’infezione da PRRSV sul sistema immunitario del suino sono ancora poco conosciuti, in quanto l’infezione e la persistenza del virus sono associate ad una complessa interazione con l’ospite, che determina una dis-regolazione dei meccanismi immunitari. Il virus presenta un tropismo selettivo per le cellule della linea monocitaria, e il principale target in vivo è rappresentato dai macrofagi alveolari. Lo scopo di questa tesi è stato quello di analizzare come viene influenzata la popolazione macrofagica in seguito all’ infezione con differenti isolati di PRRSV, sia in vitro che in vivo. Nel primo studio in vitro, il modello cellulare utilizzato è rappresentato dai “macrofagi derivanti da monociti” (monocytes derived macrophages (MDMs)), stimolati con diverse citochine quali IFNγ, IL-4 and IFNβ. Sono stati utilizzati nove diversi isolati: due Italiani, due dell’Est Europa, mentre i ceppi Lena e Lelystad sono stati utilizzati come isolati di riferimento per i ceppi ad alta e bassa patogenicità, rispettivamente. Tutti i virus utilizzati sono stati in grado di infettare il modello; una maggiore efficienza è stata riscontrata in MDMs non trattati e in quelli trattati con IL-4. Il trattamento con IFN ha determinato invece uno stato antivirale, particolarmente evidente con IFNβ. In generale, i ceppi ad alta patogenicità hanno presentato una maggiore capacità infettiva e hanno replicato a un titolo maggiore. Riguardo la produzione di citochine, IFNα and IL-10 non sono state rilevate nel surnatante delle cellule infette, mentre, HP-PRRSV PR40 è stato l’unico isolato ad indurre un significativo rilascio di TNF-α and IL1-β. Il secondo studio in vitro, è stato invece condotto utilizzando come modello cellulare i macrofagi alveolari trattati con IFNγ, IL-4 o IFNβ, per valutare la risposta all’infezione con due ceppi Italiani, un ceppo a normale patogenicità PR11 e un ceppo ad alta patogenicità PR40. Inaspettatamente, non è stata rilevata alcuna differenza tra i due isolati, indipendentemente dal trattamento dei macrofagi. Questo risultato potrebbe trovare spiegazione in una ri-polarizzazione dei macrofagi dal fenotipo M1 al fenotipo M2 determinato dall’infezione. al contrario, l’infezione di entrambi gli isolati è stata completamente bloccata dal pre-trattamento con IFNβ, sottolineando lo stato antivirale indotto dall’intereferone. Inoltre, entrambi gli isolati non hanno indotto la produzione di IFNα, IL-10 and TNF-α. Lo studio in vivo si è articolato in due diversi esperimenti, il primo ha avuto lo scopo di analizzare l’effetto indotto dall’infezione con due isolati italiani (PR11 e PR40) a diversa patogenicità, sulla popolazione macrofagica del timo, target per la replicazione del virus e per valutare la risposta immunitaria. Il secondo esperimento ha invece valutato l’efficacia di un vaccino commerciale in seguito al challenge con un ceppo ad alta patogenicita (HP-PRRSV PR40). Per valutare la risposta macrofagica all’infezione, è stata condotta un’analisi in immunoistochimica valutando la presenza del virus, TUNEL per valutare la morta cellulare e, la modulazione di diversi marker specifici per i macrofagi: CD163, CD172a, CD107a e BA4D5. Il timo degli animali infetti con entrambi i virus, PR11 e PR40 morti prima della conclusione dell’esperimento, tra i dieci e i quattordici giorni post-infezione (10-14 dpi), mostrava gravi lesioni, una maggiore presenza del virus a livello cellulare, morte cellulare ed un aumento del numero di macrofagi in tutti i compartimenti. Questi risultati suggeriscono un forte e precoce processo infiammatorio, senza alcuna particolare differenza tra gli isolati utilizzati.

Herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) have co-evolved with humans for thousands of years and are present at a high prevalence in the population worldwide. HSV infections are responsible for several illnesses including skin and mucosal lesions, blindness and even life-threatening encephalitis in both, immunocompetent and immunocompromised individuals of all ages. Therefore, diseases caused by HSVs represent significant public health burdens. Similar to other herpesviruses, HSV-1 and HSV-2 produce lifelong infections in the host by establishing latency in neurons and sporadically reactivating from these cells, eliciting recurrences that are accompanied by viral shedding in both, symptomatic and asymptomatic individuals. The ability of HSVs to persist and recur in otherwise healthy individuals is likely given by the numerous virulence factors that these viruses have evolved to evade host antiviral responses. Here, we review and discuss molecular mechanisms used by HSVs to evade early innate antiviral responses, which are the first lines of defense against these viruses. A comprehensive understanding of how HSVs evade host early antiviral responses could contribute to the development of novel therapies and vaccines to counteract these viruses.

// Chean Ring Leong 1, 4, * , Kenji Funami 1, * , Hiroyuki Oshiumi 1, 5 , Deng Mengao 1 , Hiromi Takaki 1 , Misako Matsumoto 1 , Hussein H. Aly 2 , Koichi Watashi 2 , Kazuaki Chayama 3 , Tsukasa Seya 1 1 Department of Microbiology and Immunology, Graduate School of Medicine, Hokkaido University, Sapporo, 060-8638 Japan 2 Department of Virology II, National Institute of Infectious Diseases, Tokyo, 162-8640 Japan 3 Department of Gastroenterology and Metabolism, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, 734-8553 Japan 4 Present address: Section of Bioengineering Technology, Universiti Kuala Lumpur (UNIKL) MICET, Kuala Lumpur, 78000 Malaysia 5 Present address: Department of Immunology, Graduate School of Medical Science, Kumamoto University, Kumamoto, 860-8556 Japan * These authors have contributed equally to this work Correspondence to: Tsukasa Seya, email: seya-tu@pop.med.hokudai.ac.jp Keywords: hepatitis B virus (HBV), interferon (IFN), interferon-stimulated genes (ISGs), ISG20, gene therapy Received: June 15, 2016 Accepted: August 15, 2016 Published: September 08, 2016 ABSTRACT Hepatitis B virus (HBV) barely induces host interferon (IFN)-stimulated genes (ISGs), which allows efficient HBV replication in the immortalized mouse hepatocytes as per human hepatocytes. Here we found that transfection of Isg20 plasmid robustly inhibits the HBV replication in HBV-infected hepatocytes irrespective of IRF3 or IFN promoter activation. Transfection of Isg20 is thus effective to eradicate HBV in the infected hepatocytes. Transfection of HBV genome or e-stem of HBV pgRNA (active pgRNA moiety) failed to induce Isg20 in the hepatocytes, while control polyI:C (a viral dsRNA analogue mimic) activated MAVS pathway leading to production of type I IFN and then ISGsg20 via the IFN-α/β receptor (IFNAR). Consistently, addition of IFN-α induced Isg20 and partially suppressed HBV replication in hepatocytes. Chasing HBV RNA, DNA and proteins by blotting indicated that ISG20 expression decreased HBV RNA and replicative DNA in HBV-transfected cells, which resulted in low HBs antigen production and virus titer. The exonuclease domains of ISG20 mainly participated in HBV-RNA decay. In vivo hydrodynamic injection, ISG20 was crucial for suppressing HBV replication without degrading host RNA in the liver. Taken together, ISG20 acts as an innate anti-HBV effector that selectively degrades HBV RNA and blocks replication of infectious HBV particles. ISG20 would be a critical effector for ameliorating chronic HBV infection in the IFN therapy.

Expression of adenosine deaminase acting on RNA1 (ADAR1) is driven by alternative promoters. Promoter PA, activated by interferon (IFN), produces transcripts that encode the inducible p150 ADAR1 protein, whereas PB specifies the constitutively expressed p110 protein. We show using Stat1−/−, Stat2−/− and IRF9−/− MEFs that induction of ADAR1 p150 occurs by STAT2- and IRF9-dependent signaling that is enhanced by, but not obligatorily dependent upon, STAT1. Chromatin immunoprecipitation analysis demonstrated STAT2 at the PA promoter in IFN-treated Stat1−/− cells, whereas IFN-treated wild-type cells showed both STAT1 and STAT2 bound at PA. By contrast, with human 2fTGH cells and mutants U3A or U6A, ADAR1 induction by IFN was dependent upon both STAT1 and STAT2. These results suggest that transcriptional activation of Adar1 by IFN occurs in the absence of STAT1 by a non-canonical STAT2-dependent pathway in mouse but not human cells.

Measles virus (MV) deficient in C protein (C(ko)) expression efficiently induces both stress granules (SG) and interferon (IFNβ), whereas isogenic wild-type (WT) and V mutant (V(ko)) viruses do not. We therefore examined the effect of IFNβ pretreatment on SG formation, and the roles played by the IFN-inducible double-stranded (ds) RNA-dependent protein kinase (PKR) and dsRNA adenosine deaminase (ADAR1). SG formation in ADAR1-sufficient cells infected with WT or V(ko) mutant virus was enhanced by IFN treatment and was PKR-dependent. SG formation in C(ko) virus-infected cells was already high without IFN treatment and was not further enhanced by IFN. IFN treatment alone, in the absence of infection, induced SG formation in ADAR1-deficient but not ADAR1-sufficient cells. Type I IFN-induced enhancement in SG formation occurred by a canonical IFN signaling response dependent upon STAT1 and STAT2. These results further establish ADAR1 as a suppressor of the interferon and SG innate immune responses.

Immune responses in the ovary are tightly regulated to provide protection for the developing germ cells, which are very sensitive to inflammatory responses. This characteristic immune response is often used by viral pathogens to evade the immune system, replicate and be transmitted to other specimens through the ovary. Taking into account that in teleost fish, the innate immune system is considered crucial to the outcome of viral infections and the interferon (IFN) system is considered as the first line of defence against viruses, we have studied the IFN response in rainbow trout (Oncorhynchus mykiss) ovary using two viruses with different replicative capacity in this organ, namely viral haemorrhagic septicaemia virus (VHSV) and infectious pancreatic necrosis virus (IPNV). Both VHSV and IPNV are shed from the ovary, but while VHSV actively replicates at this site, IPNV remains silent. In this context, we have determined the levels of expression of IFNs and the IFN-induced Mx genes in the ovary upon in vivo and in vitro infections with VHSV and IPNV, and compared to the effects provoked by the viral mimic poly IC in vivo. We have demonstrated that while VHSV strongly up-regulates all the IFN genes studied, IPNV in vivo exposure either has no effect or even provokes strong suppression of IFN gene expression. These differences are not observed in vitro, even though IPNV does not replicate actively in this case either. Finally, to better understand the role that the production of type I IFN plays in the ovary, we have studied the effects of two type I recombinant rainbow trout IFNs (rtIFN1 and rtIFN2) to modulate both the expression of immune genes and to establish an antiviral state in the ovary. Interestingly, the ovary was able to respond to both rtIFN1 and 2, despite the fact that the IFN1 gene was not expressed here. Moreover, rtIFN1 and rtIFN2 not only modulated the expression of genes related to the IFN response, but also modulated inflammatory genes and significantly suppressed VHSV replication. © 2010 Elsevier Ltd.

Although interferon (IFN) alpha and beta are currently recognized as the most effective agents for treating patients with chronic hepatitis B and C, they are well known to cause various adverse effects. To reduce the dose of IFN necessary for treatment, we tried enhancing the effects of IFN in the liver by ultrasound exposure. Percutaneous insonation in mouse liver following IFN-beta injection with the ultrasound power level used at clinical diagnosis enhanced the IFN-beta-induced increase of 2',5'-oligoadenylate synthetase (2-5AS) levels in the liver. This enhancement of the 2-5AS level was dependent on the duration as well as on the timing of insonation after the IFN-beta injection. In contrast, liver insonation did not enhance 2-5AS levels in the lung or spleen, and moreover, it did not alter tissue distribution of injected IFN. Thus, the combination of IFN-beta administration and subsequent liver insonation appears to be a promising method for enhancing the antiviral activity of IFN specifically in the liver, enabling a reduction in the dose necessary for treatment.

A dual immunotherapy approach employing small-molecule inhibitors of apoptosis (IAP) protein antagonists in combination with innate immune stimuli has proven to be highly synergistic and effective in animal tumor models. This strategy overcomes many of the limitations of either single agent therapy and our results suggest that the combination could be easily and effectively translated to the clinic.

Background The aims of this study were to evaluate the effects of nutritional supplementation with branched-chain amino acids (BCAA) with zinc component (Aminofeel®) on adherence to and outcome of therapy in patients treated with interferon (IFN) for chronic hepatitis C and cirrhosis and to determine whether to recommend the supplement. Methods In this retrospective study, 51 patients who received IFN therapy were investigated among 203 consecutive patients who visited our hospital and were advised regarding the potential benefit of taking Aminofeel®. Each patient was free to choose whether to purchase and take Aminofeel®. Results Twenty four patients (group 1-A) took Aminofeel® during standard IFN therapy and 13 (group 1-B) did not. Low-dose, long-term IFN (maintenance) therapy, mainly peglated (Peg)-IFN alpha 2a, was administered to 14 patients who were difficult to treat, because of no effect or harmful side effects with standard IFN therapy, and who had advanced liver fibrosis. Among the 14, 11 patients (group 2-A) took Aminofeel® and 3 (group 2-B) did not. The prevalence of obesity was significantly higher (P=0.04) in group 1-A than in group 1-B. The rate of adherence to IFN therapy was higher in group 1-A (83.3%) than in group 1-B (53.8%, P=0.05). There were no significant differences between the two groups in the rates of sustained virological response (SVR) to IFN therapy. According to multivariate analysis, two factors, SVR and intake of Aminofeel®, were associated with successful adherence to IFN therapy. The adjusted odds ratios for these two factors were 13.25 and 12.59, respectively, and each was statistically significant. The SVR rate of maintenance IFN therapy was in 18.2% group 2-A and 0% in group 2-B. Conclusion Our data show that BCAA intake is useful for adherence to and effect of IFN therapy for patients with chronic hepatitis C. Nutritional supplementation with BCAA seems to be useful for HCV-infected patients receiving IFN therapy because it is impossible to introduce standard treatment for all patients among Japan's aging population.

Gesamtdissertation, Die Schuppenflechte (Psoriasis vulgaris) zeichnet sich unter anderem durch eine Aktivierung des Interferonsystems aus. Es existieren Hinweise, dass bei Patienten mit Psoriasis eine gesteigerte zelluläre Sensitivität gegenüber der Wirkung von Interferon besteht. Um diese Hypothese zu überprüfen, ist ein geeigneter Bioassay erforderlich, mit Hilfe dessen die Sensitivität gegenüber Interferon quantitativ erfasst werden kann. Die Etablierung eines solchen Assays ist das Thema der vorliegenden Arbeit. Die Sekretion des Interferon- induzierten Proteins 10 (IP-10) wurde als Maß der Interferon-Stimulation verwendet. Wir konnten feststellen, dass die gemessene individuelle IFN Sensitivität eines Probanden unabhängig vom Anteil der CD14+ in den PBMC, dem Serum IP10 Spiegel und der maximalen IP10 Sekretion ist. Allerdings variieren die EC50 Werte der IFN induzierten IP10 Sekretion aufgrund einer schwankenden Präaktivierung der PBMC in vivo intra-individuell stark. Es konnte jedoch gezeigt werden, dass die relative Sensitivität gegenüber Typ1 und Typ2 IFN sowohl intra als auch interindividuell nur geringen intra-individuellen Schwankungen unterliegt, mithin einen stabilen individuell stabilen Interferon-Sensitivitätsindex darstellt. Diese Vermutung muss allerdings durch Messung weiterer Probanden überprüft werden. Anschließend kann der IP10 ELISA eingesetzt werden, um die IFN Sensitivität der PBMC von Psoriasispatienten mit denen von Normalpersonen zu vergleichen. Schließlich wäre eine Veränderung der IFN Sensitivität im Therapieverlauf denkbar, und mithilfe des hier entwickelten IP10 ELISA zu testen., Psoriasis has been found to be characterized by the activation of the interferon system. There is a significant indication that psoriasis patients experience increased cellular sensitivity to interferons. To validate this hypothesis we have sought to establish a bioassay to quantify patient sensitivity to interferon. We defined the interferon-inducible protein 10kDa (1P-10) to be our read-out protein. Our results showed that the interferon sensitivity of healthy probands is unaffected by the CD14+ fraction of the PBMC, the level of 1P-10 in the serum and the maximum 1P-10 secretion. However, the intraindividual variation of the IFN-induced EC50 of the 1P-10 secretion alters strongly due to a variable pre-activation of the PBMC in vivo. We were able to show that the relative sensitivity to Typ1 and Typ2 IFN varies less in an intraindividual as well as an interindividual comparison and therefore it could be used as an individually stable hallmark of the IFN sensitivity. This assumption needs to be verified in larger trials, after which the 1P-10 ELISA should be used to compare psoriasis patients with healthy probands. Finally, a possible change of the sensitivity to IFN during a therapy could be investigated with this assay.

Natural killer (NK) cells are innate immune effectors that produce various immunoregulatory cytokines. Recent studies have shown that NK cells are involved in the initiation of autoimmunity. In this study, we determined abnormalities of NK cells in systemic sclerosis (SSc), an autoimmune connective tissue disease, by assessing the frequency and absolute number, activation marker expression, cytokine production, and killing activity. The frequency and absolute number of NK cells increased in diffuse cutaneous SSc (dcSSc), whereas they were normal in limited cutaneous SSc (lcSSc). NK cells from both dcSSc and lcSSc patients exhibited activated phenotypes characterized by up-regulated CD16 and CD69 expression and downregulated CD62L expression. Interferon (IFN)-gamma production by non-stimulated NK cells from both dcSSc and lcSSc patients was increased compared to the normal control, whereas on stimulation, a reduced amount of IFN-gamma was produced. Interleukin (IL)-5 and IL-10 production by non-stimulated NK cells and IL-6 production by stimulated NK cells were augmented in dcSSc patients, but not in lcSSc patients. Despite the augmented cytokine production by non-stimulated NK cells, natural cytotoxicity activity and granzyme B secretion was reduced in NK cells from dcSSc and lcSSc patients. These results suggested that altered NK cell function contributes to immunological abnormalities in SSc.

Viral myocarditis is an important human disease, and reovirus-induced myocarditis in mice provides an excellent model to study direct viral damage to the heart. Previously, we showed that reovirus induction of and sensitivity to interferon-beta (IFN-beta) is an important determinant of viral pathogenicity in the heart and that the transcription factor interferon regulatory factor-3 (IRF-3) is required for reovirus induction of IFN-beta in primary cardiac myocyte cultures. Given several lines of evidence suggesting a possible distinctive environment for IRFs in the heart, we have now focused on IRF-1. Previous studies demonstrated that viruses, double-stranded-RNA (dsRNA), and IFN-alpha/beta can each induce IRF-1 and that IRF-1 plays a role in dsRNA, but perhaps not viral, induction of IFN-alpha/beta. Importantly, none of these studies used a virus with a dsRNA genome (such as reovirus), none of them used a highly differentiated nonlymphoid cell type, and none of them addressed whether viral induction of IRF-1 is direct or is mediated through viral induction of IFN-beta. Indeed, as recently as this year it has been assumed that viral induction of IRF-1 is direct. Here, we found that reovirus induced IRF-1 in primary cardiac myocyte cultures, but that IRF-1 was not required for reovirus induction of IFN-beta. Surprisingly, we found that reovirus failed to induce IRF-1 in the absence of the IFN-alpha/beta response. This provides the first evidence that viruses may not induce IRF-1 directly. Finally, nonmyocarditic reovirus strains induced more cardiac lesions in mice deficient for IRF-1 than they did in wildtype mice, directly demonstrating a protective role for IRF-1. Together, the results indicate that while IRF-1 is downstream of the IFN-beta response, it plays an important protective role against viral myocarditis.