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Here, we review an overall effectiveness of interferon-based preparations and interferon biosynthesis inducers for treatment of feline leukemia, as well as development of methodological approaches to improve efficacy of interferon therapy. Feline leukemia is a systemic hematopoietic malignancy caused by a single-stranded RNA retrovirus called Feline Leukemia Virus (FeLV) that leads to lethal outcome within about 3 years after the onset. FelV is widely distributed in population of domestic cats worldwide, being often detected in the blood of wild cats, including those of rare and endangered species. In some regions, FeLV prevalence may be high not only among domestic cats, but among wild as well. Currently, there are several commercially available vaccines to protect cats from FeLV infection (e.g., inactivated whole-virion vaccines such as Nobivac adjuvanted Feline 2-FeLV, two-adjuvant subunit vaccine FeLV-derived protein antigens as well as non-adjuvanted vector DNA vaccine). However, none of such vaccines provides durable protection. In addition, vaccination of cats against FeLV is often associated with development of diverse inflammatory, allergic and shock complications, and highly serious side effects such as developing vaccine-associated sarcoma at the injection site that some researchers connect with use of adjuvants like aluminum salts etc. We briefly describe FeLV virus, pathogenetic parametersassociated with FeLV infection as well as current technologies for preventing and treating feline leukemia. A historic background and current state of interferon therapy for FeLV infection as well as associated neoplastic processes in domestic cats and some wild species are evaluated. Possible interventions aimed at improving efficiency of interferon therapy of feline leukemia based on using new recombinant interferon preparations of various types and subtypes, as well as interferon inducers are discussed. In conclusion, it is noted that another interesting and potentially highly promising option in defining strategy of biotherapy associated with modulating IFN system in FeLV-infected animals might be to use of synthetic inducers triggering endogenous IFN production.

Type 1 interferon (IFN) is a major contributor to the pathogenesis of systemic lupus erythematosus (SLE). A landmark study by Caielli et al. now shows that erythrocytes from lupus patients that fail to switch from glycolysis to oxidative phosphorylation during differentiation retain their mitochondria. These mitochondria-containing erythrocytes represent a novel source of IFN when phagocytosed by macrophages.

The aim of the study was to assess the antiviral activity of the 7 types of liquid corneal storage medium on an experimental model of herpesvirus infection in vitro. Material and methods. The study of antiviral activity of 7 samples of liquid corneal storage medium on a model of herpesvirus infection caused by the herpes simplex virus type 1 in Vero-cell using virological and statistical research methods was carried out. Results and discussion . All the studied images of the corneal storage medium, including the Borzenka-Moroz base medium, did not have a cytotoxic effect on Vero cell culture. Out of of 7 samples of liquid corneal storage medium, 4 samples had reliable antiviral activity against HSV-1 when used under the therapeutic regimen (1 hour after infection) and under the preventive regimen (2 hours before infection). Antiviral activity was established in 2 samples containing the interferon inducer cycloferon at a concentration of 10 mg/kg and 30 mg/kg (sample 2, 3), in a sample containing the interferon inducer gamapren 15 mg/kg (sample 5), and in a sample containing a combination of drugs - 10 mg/kg cycloferon and an acyclic nucleoside analog-acyclovir 10 mg/kg (sample 6). According to the results of 2 test regimens, the maximum statistically significant inhibitory effect in relation to HSV-1 was detected in sample 6, containing a combination of drugs. Against the background of sample 6, the infectious activity of the test virus decreased by an average of 3.2 lg, the inhibition coefficient was 54.5%. Conclusion. The results of the study indicate the prospects of using types of media with antiviral activity (samples 2, 3, 5, 6) for storing donor corneas in order to increase the effectiveness of keratoplasty in patients with ophthalmic herpes.

Despite the obvious advances in vaccination and therapy, influenza remains a poorly controlled infection with high morbidity and mortality. This study examined the antiviral activity of interferon inducer Kagocel on a mouse model of lethal influenza pneumonia. It has been shown that the therapeutic and prophylactic use of Kagocel leads to a dose-dependent decrease in specific mortality and suppression of virus reproduction in lung tissue. The effect of Kagocel was statistically identical to the effect of the reference drug — Arbidol (umifenovir).

Introduction. Interferons (IFN) and IFN inducers are effective in suppressing viral reproduction and correcting of the innate immunity mechanisms. The aim of the study was to test the hypothesis of the possible involvement of the IFN inducer CelAgrip (CA) as an activator or suppressor of antiviral effects in Burkitt’s lymphoma (LB) cell cultures with different ability to produce Epstein-Barr virus antigens (EBV). Material and methods. The kinetic analysis of the dynamics of reactive oxygen species (ROS) production and determination of gene group expression by real-time PCR in response to CA treatment were done in human cell lines LB P3HR-1 and Namalva, spontaneously producing and not producing EBV antigens. Results and discussion. When treating CA in Namalva cells, a decrease in the ROS activation index was found; in P3HR-1 cells, an increase was observed. After treatment with CA, there was no reliable activation of the IFN-α, IFN-β and IFN-λ genes in Namalva cells, but the expression of the ISG15 and P53(TP53) genes was increased more than 1200 times and 4.5 times, respectively. When processing the CA of P3HR-1 cells, the expression of IFN-α genes increased by more than 200 times, IFN-λ - 100 times, and the ISG15 gene - 2.2 times. The relationship between IFN-inducing action of CA and the activity of ISG15 and ROS in LB cell cultures producing and not producing EBV antigens is supposed. Conclusion. In Namalva cells that do not produce EBV antigens the treatment of CA results in suppression of ROS generation and activation of the expression of genes ISG15 and P53 (TP53); in P3HR-1 cells producing EBV antigens, the opposite picture is observed - the formation of ROS and the expression of the IFN-α and IFN-λ genes are activated and the activity of the ISG15 and P53 (TP53) genes is suppressed.

To assess the effectiveness of the use of the antiviral drug enisamium iodide in the complex treatment of acute respiratory viral infections (ARVI) caused by various pathogens in routine clinical practice.А prospective randomized study included 134 patients who were treated in the epidemic season of influenza and ARVI in 20182019. All patients were examined for the presence of influenza A and B viruses, respiratory syncytial virus, human metapneumovirus, parainfluenza virus, coronaviruses, rhinoviruses, adenoviruses in nasopharyngeal swabs by PCR. Patients of the main group received enisamium iodide along with symptomatic therapy, the control group received only symptomatic therapy. The primary parameter of the effectiveness of therapy was evaluated on the scale of the general severity of the manifestations of ARVI (Total Symptom Score TSS) from the 2nd to the 4th day and by the secondary criteria of effectiveness: assessment of the duration of ARVI, the severity of fever, the proportion of patients with normal body temperature, the duration of the main clinical symptoms of acute respiratory viral infections, the proportion of patients in whom complications requiring antibiotics were noted, the dynamics of interferon status on the 6th day. To conduct a statistical analysis, depending on the efficiency parameter, the ANCOVA method with a fixed group factor and an initial score on the TSS severity scale was used as covariates, a criterion for comparing quantitative indicators in two independent groups.According to the results of the analysis of the primary efficacy parameter, the median (interquartile range) of the average score on the scale of the general severity of ARVI manifestations in the main group was 4.33 (3.675.83), in the comparison group 6.00 (4.677.25; p0.001). The duration of systemic and local manifestations of acute respiratory viral infections was statistically significantly less in the main group (p=0.002 and p=0.019, respectively). Prescription of additional therapy was required in 2 (2.9%) patients of the main group (patients taking enisamium iodide), compared with 8 (11.9%) patients in the control group. Serum levels of interferon  and interferon  on the last day of treatment were statistically significantly higher in patients of the main group compared with the control group (p0.001). Treatment (excellent) was evaluated by 42 (62.7%) patients, while in the control group only 17 (25.8%) patients gave similar ratings. Both patients (p0.001) and doctors (p0.002) rated therapy tolerance better in the study group.The results confirmed the safety and effectiveness of enisamium iodide as a treatment for ARVI and influenza. The antiviral, interferonogenic and anti-inflammatory properties of the drug are involved in the formation of an antiviral response and reduce the risk of complications, which makes it possible to reduce the number of symptomatic agents used.Цель. Оценка эффективности применения в рутинной клинической практике противовирусного препарата энисамия йодида в комплексной терапии острых респираторных вирусных инфекций (ОРВИ), вызванных различными возбудителями. Материалы и методы. В проспективное рандомизированное исследование включены 134 пациента, проходивших лечение в эпидемический сезон гриппа и ОРВИ в 20182019 гг. Все пациенты обследованы на наличие вирусов гриппа А и B, респираторно-синцитиального вируса, метапневмовируса человека, вируса парагриппа, коронавирусов, риновирусов, аденовирусов в мазках из носоглотки методом полимеразной цепной реакции. Пациенты основной группы получали энисамия йодид наряду с симптоматической терапией, контрольная группа получала только симптоматическую терапию. Первичный параметр эффективности лечения оценивали по шкале общей выраженности проявлений ОРВИ (Total Symptom Score TSS) со 2 по 4-й день и вторичным критериям эффективности: оценка продолжительности ОРВИ, выраженность лихорадки, доля пациентов с нормальной температурой тела, продолжительность основных клинических симптомов ОРВИ, доля больных, у которых отмечено развитие осложнений, требующих назначения антибиотиков, динамика интерферонового статуса на 6-й день. Для проведения статистического анализа в зависимости от параметра эффективности применялся метод ANCOVA с фиксированным фактором группы и исходным баллом по шкале выраженности TSS в качестве ковариаты, критерий сравнения количественных показателей в двух независимых группах. Результаты. По результатам анализа первичного параметра эффективности медиана (межквартильный диапазон) средней оценки по шкале общей выраженности проявлений ОРВИ в основной группе составила 4,33 (3,675,83), в группе сравнения 6,00 (4,677,25; p0,001. Продолжительность системных и локальных проявлений ОРВИ была статистически значимо меньше в основной группе (р=0,002 и р=0,019 соответственно). Назначение дополнительной терапии потребовалось 2 (2,9%) пациентам основной группы (принимающие энисамия йодид) по сравнению с 8 (11,9%) пациентами из контрольной группы. Сывороточные уровни интерферона  и интерферона  на последний день лечения были статистически значимо выше у пациентов основной группы по сравнению с контрольной (p0,001). Оценку лечению отлично дали 42 (62,7%) пациента, в то время как в контрольной группе аналогичные оценки поставили только 17 (25,8%) больных. И пациенты (p0,001), и врачи (p0,002) оценивали переносимость терапии лучше в основной группе. Заключение. Полученные результаты подтвердили безопасность и эффективность энисамия йодида как средства терапии ОРВИ и гриппа. Противовирусные, интерфероногенные и противовоспалительные свойства препарата участвуют в формировании противовирусного ответа и снижают риск осложнений, что дает возможность уменьшить количество применяемых симптоматических средств.

Significance Here, we show that expression of key circadian clock genes in the skin is altered by acute inflammation in mice treated topically with the immune activator imiquimod and by chronic inflammation in human psoriatic lesions. We show time-of-day–dependent activation of the interferon pathway, a key pathway involved in the host defense response. Mice lacking circadian rhythms have greater epidermal hyperplasia and more robust activation of the interferon pathway. Furthermore, we show that daytime-restricted feeding shifts the phase of interferon-sensitive gene expression in mouse skin. These findings demonstrate a role for the circadian clock in a major defense pathway in the skin’s response to microbes and cancer and suggest that timing of feeding can affect this response in the skin., The circadian clock coordinates a variety of immune responses with signals from the external environment to promote survival. We investigated the potential reciprocal relationship between the circadian clock and skin inflammation. We treated mice topically with the Toll-like receptor 7 (TLR7) agonist imiquimod (IMQ) to activate IFN-sensitive gene (ISG) pathways and induce psoriasiform inflammation. IMQ transiently altered core clock gene expression, an effect mirrored in human patient psoriatic lesions. In mouse skin 1 d after IMQ treatment, ISGs, including the key ISG transcription factor IFN regulatory factor 7 (Irf7), were more highly induced after treatment during the day than the night. Nuclear localization of phosphorylated-IRF7 was most prominently time-of-day dependent in epidermal leukocytes, suggesting that these cell types play an important role in the diurnal ISG response to IMQ. Mice lacking Bmal1 systemically had exacerbated and arrhythmic ISG/Irf7 expression after IMQ. Furthermore, daytime-restricted feeding, which affects the phase of the skin circadian clock, reverses the diurnal rhythm of IMQ-induced ISG expression in the skin. These results suggest a role for the circadian clock, driven by BMAL1, as a negative regulator of the ISG response, and highlight the finding that feeding time can modulate the skin immune response. Since the IFN response is essential for the antiviral and antitumor effects of TLR activation, these findings are consistent with the time-of-day–dependent variability in the ability to fight microbial pathogens and tumor initiation and offer support for the use of chronotherapy for their treatment.

BackgroundViral-based immunotherapy can overcome resistance to immune checkpoint blockade (ICB) and fill the unmet needs of many patients with cancer. Oncolytic viruses (OVs) are defined as engineered or naturally occurring viruses that selectively replicate in and kill cancer cells. OVs also induce antitumor immunity. The purpose of this study was to compare the antitumor effects of live oncolytic vaccinia viruses versus the inactivated versions and elucidate their underlying immunological mechanisms.MethodsWe engineered a replication-competent, oncolytic vaccinia virus (OV-GM) by inserting a murine GM-CSF gene into the thymidine kinase locus of a mutant vaccinia E3L∆83N, which lacks the Z-DNA-binding domain of vaccinia virulence factor E3. We compared the antitumor effects of intratumoral (IT) delivery of live OV-GM versus heat-inactivated OV-GM (heat-iOV-GM) in a murine B16-F10 melanoma bilateral implantation model. We also generated vvDD, a well-studied oncolytic vaccinia virus, and compared the antitumor effects of live vvDD vs heat-inactivated vvDD (heat-ivvDD) in a murine A20 B-cell lymphoma bilateral tumor implantation model.ResultsHeat-iOV-GM infection of dendritic cells (DCs) and tumor cells in vitro induced type I interferon and proinflammatory cytokines and chemokines, whereas live OV-GM did not. IT live OV-GM was less effective in generating systemic antitumor immunity compared with heat-iOV-GM. Similar to heat-iOV-GM, the antitumor effects of live OV-GM also require Batf3-dependent CD103+ dendritic cells. When combined with systemic delivery of ICB, IT heat-iOV-GM was more effective in eradicating tumors, compared with live OV-GM. IT heat-ivvDD was also more effective in treating murine A20 B-cell lymphoma, compared with live vvDD.ConclusionsTumor lysis induced by the replication of oncolytic vaccinia virus has a limited effect on the generation of systemic antitumor immunity. The activation of Batf3-dependent CD103+ DCs is critical for antitumor effects induced by both live OV-GM and heat-iOV-GM, with the latter being more potent than live OV-GM in inducing innate and adaptive immunity in both locally injected and distant, non-injected tumors. We propose that evaluations of both innate and adaptive immunity, induced by IT oncolytic viral immunotherapy at injected and non-injected tumors, should be included as potential biomarkers for host responses to viral therapy.

Acute respiratory viral infections (ARVI) are some of the most widely spread diseases worldwide that are of 80% of economic cost of infectious diseases. It is known that specific prophylaxis (vaccination) and, in some clinical cases, direct-acting antiviral prophylaxis are key strategies for the prevention of influenza as one of the most dangerous ARVI viruses. However, there are a number of reasons why specific prophylaxis of influenza does not entirely address the issue of prophylaxis of ARVI that are caused by more than 200 different viruses, such as formation of immune response only to vaccine influenza strains and absence of immune response to other respiratory infections, induction in the case of short-term protection, especially, in the aged and others. Prevention of influenza with antiviral drugs is limited by risks of development of resistant influenza strains aa well as contraindications and limitations of use. Therefore, the use of nonspecific drugs such as broad-spectrum antiviral drugs of interferon type (IFN) and their inducers (II), as well as sanitary and hygienic measures is recommended for prevention. For the purpose of prevention, the use of inducers of endogenic interferons is relevant in unvaccinated persons during pre-epidemic and epidemic ARVI periods; in high-risk group (students, medical professions, transport workers, elderly, patients with comorbid pathologies, people in structured organizations, for example, military personnel), in immunocompromised people. One of the most extensively studied interferon inducers widely used in clinical practice since 2003 for prevention and treatment of influenza and diseases caused by Herpesviruses is an antiviral drug Kagocel®. The paper presents real-time data of the preventive efficacy of Kagocel® in the therapy of acute respiratory viral infections and influenza caused by different viral strains including pandemic ones.

Introduction . Cytokines activated in response to immunosuppressive viral infections can directly or indirectly affect the neoplastic transformation of B cells. In this study, we studied a new substance designed to produce the antiviral drug CelAgrip (CA, CelAgripus), which exhibits interferon (IFN) and cytokine-inducing activity and, apparently, can be used as an activator of antiviral immunity. Purpose - is to evaluate the cytokine-regulating effect of CA in Burkitt's lymphoma (LB) cell lines latently infected with the Epstein-Barr virus (EBV). Objectives : to study the CA-induced expression of the cytokine genes IL-ip, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-a, IFN -Y, IFN-p, IFN-A1, IFN-A2, IFN-A3, TNF-a in normal and EBV transformed LB cells. Material and methods . Cell line: the human embryo fibroblasts (HEF), Namalva, Daudi, Raji, P3HR-1. Preparations: CA, gossypol-acetic acid (GAA), sodium carboxymethyl cellulose (Na-CMC). Methods: RT-PCR and methods for assessing cytotoxicity (MTT and Scepter 2.0 Merck cell counter). Results . The effect of the CA preparation on the expression of IFN- Л , IL-1p, IL-6, IL-8 and IL-10 genes was revealed. Discussion. We observed the activation of gene expression of IFN-A, IL-1P, IL-6, IL-8 and suppression of IL-10 gene activity when treatment CA of LB cells. Conclusion . The substance CA has new effects on the activation of the expression of a number of key cytokine genes in stable Burkitt lymphoma cell lines.

In this article topical issues in treatment of acute respiratory infections in children and place of both exogenous interferons and interferon inducer drugs in their treatment are reviewed, including mechanisms of action, advantages and disadvantages of their application, special features of use in children. The latest clinical and laboratory data on the efficacy meglumine acridone acetate use in this pathology are given here. Depletion of inflammatory manifestations, correction of immune system imbalance were observed, leading to reduction of symptoms severity and disease duration, which, along with the good tolerability of the drug allows us to recommend it for widespread use in children.

Immunotherapy has revolutionized cancer treatment, with sustained responses to immune checkpoint inhibitors reported in a number of malignancies. Such therapeutics are now being trialed in aggressive or advanced cancers that are heavily reliant on untargeted therapies, such as triple negative breast cancer. However, responses have been underwhelming to date and are very difficult to predict, leading to an inability to accurately weigh up the benefit-to-risk ratio for their implementation. The tumor immune microenvironment has been closely linked to immunotherapeutic response, with superior responses observed in patients with T cell-inflamed or 'hot' tumors. One class of cytokines, the type I interferons, are a major dictator of tumor immune infiltration and activation. Tumor cell inherent interferon signaling dramatically influences the immune microenvironment and the expression of immune checkpoint proteins, hence regulators and targets of this pathway are candidate biomarkers of immunotherapeutic response. In support of a link between IFN signaling and immunotherapeutic response, the combination of type I interferon inducers with checkpoint immunotherapy has recently been demonstrated critical for a sustained anti-tumor response in aggressive breast cancer models. Here we review evidence that links type I interferons with a hot tumor immune microenvironment, response to checkpoint inhibitors and reduced risk of metastasis that supports their use as biomarkers and therapeutics in oncology.

Bronchial asthma (BA) remains one of the most pressing problems in modern pediatrics. The incidence of this pathology is increasing annually and, according to WHO, about 300 million people in the world suffer from asthma. Despite the recent advances in the diagnosis and treatment of asthma, complete control over the course of asthma has failed. The objective of the workwas to evaluate the effect of immunomodulatory therapy on nonspecific and humoral immunity in children with BA by including interferon inducer (cycloferon) in a standard base therapy. There were 120 children with bronchial asthma aged 5 to 14 years, with an average age of (11.6 ± 1.5) years. To determine the diagnosis and effectiveness of therapy used the international classification of diseases of the 10th examination, protocol for the diagnosis and treatment of bronchial asthma in children (order of the Ministry of Health of Ukraine № 767 from 27.12.2005), GINA 2011. The main group (n = 60) included children, who, in addition to basic therapy, received in addition interferon inducer (cycloferon) according to the scheme: 150 mg on 1-, 2-, 4-, 6- and 8-th day of therapy (No. 5) and then 150 mg after 72 hours. (No. 5) (1500 mg total). The comparison group included children (n = 60) who received only basic therapy, depending on the severity of the disease. The control group consisted of 25 healthy children. The level of serum interferon, virus-induced production of interferon-a (IFN-a), mitogen-stimulated production of interferon-g (IFN-g), the level of total serum IgE were determined by the enzyme-linked immunosorbent assay. The phagocytic activity of neutrophils (NG) was evaluated. The circulating immune complexes (CECs), immunoglobulins (Ig) A, M and G in serum were determined by the immunodiffusion reaction method. It was found that the additional use of cycloferon had a corrective effect on phagocytes, which led to an increase in the synthesis of IFN-a and IFN-g, a decrease in the severity of the allergic component due to the decrease in IgE and circulating immune complexes and a normalization of the synthesis of IgA, IgM, IgG with further development of long-term clinical remission of the disease.

The article justifies the use of interferon inducers in a clinical practice of pediatrician. The authors isolated a group of derivatives of acridone acetic acid, which effectiveness is associated with the features of pharmacokinetics and pharmacological activity. They demonstrated preventive and therapeutic efficacy of cycloferon in the complex treatment of children with influenza and other acute respiratory viral infections, herpes virus, arbovirus, rotavirus infections, viral hepatitis and tick-borne encephalitis due to its immunomodulatory potency. The article analyzed the use of cycloferon in frequently sick children, patients with allergic diseases, chronic tonsillitis, etc.; also it showed a decrease in the frequency of relapses of both infectious and allergic diseases, the possibility of combined use with other etiotropic and symptomatic drugs.

The aim of the study was to evaluate the ARVI prevention effectiveness in patients with chronic heart failure (CHF) using interferon inducer amixin. Materials and methods. Conducted a comprehensive survey, dynamic monitoring and treatment of 60 patients aged from 49 to 70 years (mean age 60.25±4.57 years, 17 men and 43 women) with CHF with preserved ejection fraction of left ventricle (LVEF) (≥50%), II-III functional class (FC) according to the classification of new York Heart Association (NYHA), which developed as a result of coronary heart disease (CHD), hypertensive disease (HD). Of these, 30 patients (group 1) on the background of standard therapy for CHF received for the prevention of ARVI tiloron (Amixin) at a dose of 125 mg once a week for 6 weeks, two courses for 1 year. Group 2 patients received only standard therapy for CHF. Results. A decrease in the frequency of ARVI in patients with CHF treated with Amixin was found, which was accompanied by a decrease in the severity of subclinical inflammation by reducing the production of proinflammatory (IL-1β) and increasing the production of anti-inflammatory (IL-10) cytokines, reducing neurohumoral activation (reducing levels of aldosterone and Nt-proBNP), increasing the level of α- and γ-interferon. The positive dynamics of biomarkers of systemic inflammation and neurohormonal activation explains the improvement of the clinical course in patients with CHF (increase of tolerance to physical loads, reducing the number of visits to General practitioner and hospital admissions in the hospital during 12 months of observation). Conclusion. A promising approach to the prevention of SARS in patients with CHF is course therapy with Amixin (2 times a year before the seasonal rising in the incidence of respiratory viral infections and influenza), which allows to achieve both decreasing in the frequency of SARS per year, and improvement the clinical course of CHF.

Early growth response gene (EGR1) is a transcription factor known to be a downstream effector of B-cell receptor signaling and Janus kinase 1 (JAK1) signaling in diffuse large B-cell lymphoma (DLBCL). While EGR1 is characterized as a tumor suppressor in leukemia and multiple myeloma, the role of EGR1 in lymphoma is unknown. Here we demonstrate that EGR1 is a potential oncogene that promotes cell proliferation in DLBCL. IHC analysis revealed that EGR1 expression is elevated in DLBCL compared with normal lymphoid tissues and the level of EGR1 expression is higher in activated B cell–like subtype (ABC) than germinal center B cell–like subtype (GCB). EGR1 expression is required for the survival and proliferation of DLBCL cells. Genomic analyses demonstrated that EGR1 upregulates expression of MYC and E2F pathway genes through the CBP/p300/H3K27ac/BRD4 axis while repressing expression of the type I IFN pathway genes by interaction with the corepressor NAB2. Genetic and pharmacologic inhibition of EGR1 synergizes with the BRD4 inhibitor JQ1 or the type I IFN inducer lenalidomide in growth inhibition of ABC DLBCL both in cell cultures and xenograft mouse models. Therefore, targeting oncogenic EGR1 signaling represents a potential new targeted therapeutic strategy in DLBCL, especially for the more aggressive ABC DLBCL. Implications: The study characterizes EGR1 as a potential oncogene that promotes cell proliferation and defines EGR1 as a new molecular target in DLBCL, the most common non-Hodgkin lymphoma.

Interferon-stimulated genes (ISGs) create multiple lines of defense against viral infection. Here, we show that interferon-induced protein 35 (IFI35) inhibits swine (H3N2) influenza virus replication by directly interacting with the viral protein NS1. IFI35 binds more preferentially to the effector domain of NS1 (amino acids 128 to 207 [128-207aa]) than to the viral RNA sensor RIG-I. This promotes mutual antagonism between IFI35 and NS1 and frees RIG-I from IFI35-mediated K48-linked ubiquitination and degradation. However, IFI35 does not interact with the NS1 encoded by avian (H7N9) influenza virus, resulting in IFI35 playing an opposite virus-enabling role during highly pathogenic H7N9 virus infection. Notably, replacing the 128-207aa region of H7N9 NS1 with the corresponding region of H3N2 NS1 results in the chimeric NS1 acquiring the ability to bind to and mutually antagonize IFI35. IFI35-deficient mice accordingly exhibit more resistance to lethal H7N9 infection than exhibited by their wild-type control. Our data uncover a novel mechanism by which IFI35 regulates RIG-I-mediated antiviral immunity through mutual antagonism with influenza virus protein NS1. IMPORTANCE Influenza A virus (IAV) infection poses a global health threat and is among the most common contagious pathogens to cause severe respiratory infections in humans and animals. ISGs play a key role in host defense against IAV infection. In line with others, we show IFI35-mediated ubiquitination of RIG-I to be involved in innate immunity. Moreover, we define a novel role of IFI35 in regulating the type I interferon (IFN) pathway during IAV infection. We found that IFI35 regulates RIG-I-mediated antiviral signaling by interacting with IAV NS1. H3N2 NS1, but notably not H7N9 NS1, interacts with IFI35 and efficiently suppresses IFI35-dependent ubiquitination of RIG-I. IFI35 deficiency protected mice from H7N9 virus infection. Therefore, manipulation of IFI35-NS1 provides a new approach for the development of anti-IAV treatments.

The world is now entering its 9th month of combat against a pandemic of deadly pneumonia. Started out from China in December 2019, the disease has been declared as caused by infection with a so far unknown RNA Coronavirus of the respiratory family, then named severe acute respiratory syndrome coronavirus (SARS)-CoV-2. In the absence of a vaccine, and with scientists still struggling for an effective therapy, COVID-19 (the SARS-dependent syndrome) carries up to now, a death toll of more than 590,000 (July 18,2020) undermining jobs and finance of contemporary society in all continents. Social distancing, the only measure hitherto shown to restrain virus spread, has been progressively loosened from May 2020 in some countries, leaving us in the fear of repeat attacks from the unchecked virus. We discuss the problem and propose to tentatively boost the anti-virus cell machinery by using lab-made viral mimics to engage cell receptors.

BackgroundEndogenous retroviruses (ERVs) play a role in a variety of biological processes, including embryogenesis and cancer. DNA methyltransferase inhibitors (DNMTi)-induced ERV expression triggers interferon responses in ovarian cancer cells via the viral sensing machinery. Baseline expression of ERVs also occurs in cancer cells, though this process is poorly understood and previously unexplored in epithelial ovarian cancer (EOC). Here, the prognostic and immunomodulatory consequences of baseline ERV expression was assessed in EOC.MethodsERV expression was assessed using EOC transcriptional data from The Cancer Genome Atlas (TCGA) and from an independent cohort (Hammersmith Hospital, HH), as well as from untreated or DNMTi-treated EOC cell lines. Least absolute shrinkage and selection operator (LASSO) logistic regression defined an ERV expression score to predict patient prognosis. Immunohistochemistry (IHC) was conducted on the HH cohort. Combination of DNMTi treatment with γδ T cells was tested in vitro, using EOC cell lines and patient-derived tumor cells.ResultsERV expression was found to define clinically relevant subsets of EOC patients. An ERV prognostic score was successfully generated in TCGA and validated in the independent cohort. In EOC patients from this cohort, a high ERV score was associated with better survival (log-rank p=0.0009) and correlated with infiltration of CD8+PD1+T cells (r=0.46, p=0.0001). In the TCGA dataset, a higher ERV score was found in BRCA1/2 mutant tumors, compared to wild type (p=0.015), while a lower ERV score was found in CCNE1 amplified tumors, compared to wild type (p=0.019). In vitro, baseline ERV expression dictates the level of ERV induction in response to DNMTi. Manipulation of an ERV expression threshold by DNMTi resulted in improved EOC cell killing by cytotoxic immune cells.ConclusionsThese findings uncover the potential for baseline ERV expression to robustly inform EOC patient prognosis, influence tumor immune infiltration and affect antitumor immunity.

The mitochondrial antiviral-signaling protein (MAVS, also known as VISA, IPS-1, or CARDIF) plays an essential role in the type I interferon (IFN) response and in retinoic acid-inducible gene I (RIG-I) mediated antiviral innate immunity in mammals. In this study, the caprine MAVS gene (caMAVS, 1566 bp) was identified and cloned. The caMAVS shares the highest amino acid similarity (98.1%) with the predicted sheep MAVS. Confocal microscopy analysis of partial deletion mutants of caMAVS revealed that the transmembrane and the so-called Non-Characterized domains are indispensable for intracellular localization to mitochondria. Overexpression of caMAVS in caprine endometrial epithelial cells up-regulated the mRNA levels of caprine interferon-stimulated genes. We concluded that caprine MAVS mediates the activation of the type I IFN pathway. We further demonstrated that both the CARD-like domain and the transmembrane domain of caMAVS were essential for the activation of the IFN-β promotor. The interaction between caMAVS and caprine RIG-I and the vital role of the CARD and NC domain in this interaction was demonstrated by co-immunoprecipitation. Upon infection with the Peste des Petits Ruminants Virus (PPRV, genus Morbillivirus), the level of MAVS was greatly reduced. This reduction was prevented by the addition of the proteasome inhibitor MG132. Moreover, we found that viral protein V could interact and colocalize with MAVS. Together, we identified caMAVS as a RIG-I interactive protein involved in the activation of type I IFN pathways in caprine cells and as a target for PPRV immune evasion.

Introduction . Medicines from the group of interferon inducers (IFNs) “swith on” the synthesis of type 1 interferons (IFN-I) and induce the expression of IFN-stimulated genes (ISGs) that regulate innate immunity reactions and protect the host from infectious agents and the tumour pathology. The purpose of the study was to determine the role of the drug celagrip (CA) in the activation of innate immunity genes and the effect on the production of reactive oxygen species (ROS) in patients with follicular lymphoma (FL). Objectives : to study the intensity of ROS production and the level of expression of the IFN-α2, IFN-λ1, ISG15, BCL2, P53 (TP53) and USP18 genes in response to the treatment of blood cells of patients with FL with the preparation of CA. Material and methods . The study involved primary cancer patients diagnosed with follicular lymphoma (FL) and healthy volunteers. A kinetic analysis of the dynamics of production of reactive oxygen species (ROS) was performed in whose blood cells, and the expression of the group of genes was determined by real-time PCR in response to CA processing. Results and discussion . ROS production by blood cells of patients with FL and volunteers in the presence of CA significantly decreased ( P < 0.05). The level of gene expression of ISG15 , P53 (TR53) and USP 18 in the group of patients with FL was significantly higher than that in the group of volunteers. When treating blood cells with CA, it becomes possible to divide patients with FL into groups with a positive and negative response in accordance with the level of expression of the USP18 gene. We divided FL patients into groups with a positive and negative response in accordance with the level of USP18 gene expression after treatment of blood cells with CA. Conclusions . The CA drug reduces the production of ROS and simultaneously stimulates the activity of the innate immunity genes ISG15 , P53( TP53 ) and USP18 in the blood cells of patients with FL.

Graphical abstract, Highlight • We designed and synthesized a series of arylpiperazine derivatives as interferon inducers. • We tested different linker system and carbonyl moiety resulted in increase of potency. • 5i exhibited the activity of interferon inducer with EC50 of 13.1 μM and no cytotoxicity. • 5i initiated immune responses by mediating type I IFN signaling., Type I Interferon (IFN) signaling plays an important role in the immune defense system against virus infection and in the innate immune response, thus IFNs are widely used as anti-viral agents and treatment for immune disorder or cancer. However, there is a growing demand for novel small-molecule IFN inducer due to tolerance, toxicity, or short duration of action following direct administration of IFNs. In this study, we assessed arylpiperazine (ARP) as a new core skeleton of IFN inducer. To investigate structure-activity relationship, we designed and synthesized a series of ARP analogues and evaluated the ability to stimulate IFN response in THP-1 human monocyte cells. Compound 5i was identified as a potent type I IFN inducer as it significantly increased cytokine secretion and increased expression of various IFN-stimulating genes which are representative biomarkers of type I IFN pathway. Our results suggested a beneficial therapeutic potential of 5i as an anti-viral agent.

High expression of suppressors of cytokine signalling (SOCS) has been detected during various viral infections. As a negative feedback regulator, SOCS participates in the regulation of multiple signalling pathways. In this study, to study the related mechanism between SOCS and BDV and to explore the effect of SOCS on IFN pathways in nerve cells, downregulated of SOCS1/3 in oligodendroglial (OL) cells and OL cells persistently infected with BDV (OL/BDV) were constructed with RNA interference technology. An interferon inducer (poly I:C, PIC) and an IFN-α/β R1 antibody were used as stimulation in the SOCS1/3 low-expression cell models, qRT-PCR was used to detect type I IFN and BDV nucleic acid expression, Western blot was used to detect the expression of BDV P40 protein. After BDV acute infection with OL cells which with downregulated SOCS expression, the virus accounting was not detected, and the viral protein expression was lower than that of OL/BDV cells; the OL/BDV cells with downregulated SOCS expression had lower virus nucleic acid and protein expression than OL/BDV cells. Stimulated by IFN-α/β R1 antibody, the expression of type I interferon in OL/BDV cells decreased, and the content of BDV nucleic acid and protein increased, which was higher than that of OL/BDV cells. From the results, it was concluded that downregulating SOCS1/3 can inhibit the formation of acute BDV infection and virus replication in persistent BDV infection by promoting the expression of IFN-α/β and that SOCS can be used as a new target for antiviral therapy.

BackgroundMitochondrial Lon is a chaperone and DNA-binding protein that functions in protein quality control and stress response pathways. The level of Lon regulates mitochondrial DNA (mtDNA) metabolism and the production of mitochondrial reactive oxygen species (ROS). However, there is little information in detail on how mitochondrial Lon regulates ROS-dependent cancer immunoescape through mtDNA metabolism in the tumor microenvironment (TME).MethodsWe explored the understanding of the intricate interplay between mitochondria and the innate immune response in the inflammatory TME.ResultsWe found that oxidized mtDNA is released into the cytosol when Lon is overexpressed and then it induces interferon (IFN) signaling via cGAS-STING-TBK1, which upregulates PD-L1 and IDO-1 expression to inhibit T-cell activation. Unexpectedly, upregulation of Lon also induces the secretion of extracellular vehicles (EVs), which carry mtDNA and PD-L1. Lon-induced EVs further induce the production of IFN and IL-6 from macrophages, which attenuates T-cell immunity in the TME.ConclusionsThe levels of mtDNA and PD-L1 in EVs in patients with oral cancer function as a potential diagnostic biomarker for anti-PD-L1 immunotherapy. Our studies provide an insight into the immunosuppression on mitochondrial stress and suggest a therapeutic synergy between anti-inflammation therapy and immunotherapy in cancer.

The COVID-19 pandemic has killed over 400 000 people globally. Ecological evidence indicates that countries with national universal BCG vaccination programs for tuberculosis (TB) prevention have a much lower incidence of severe COVID-19 and mortality compared with those that do not have such programs. BCG is a century old vaccine used for TB prevention via infant/childhood vaccination in lowto middle-income countries with high infection prevalence rate and is known to reduce all-cause neonatal mortality. BCG remains the standard immunotherapy treatment for patients with high-risk non-muscle invasive bladder cancer globally for more than 44 years. Several trials are, therefore, investigating BCG as a prophylactic against COVID-19 in healthcare workers and the elderly. In this commentary, we discuss the potential mechanisms that may underlie BCG associated heterologous protection with a focus on tertiary lymphoid structure (TLS) organogenesis. Given the significance of TLSs in mucosal immunity, their association with positive prognosis and response to immune checkpoint blockade with a critical role of Type I interferon (IFN-1) in inducing these, we also discuss potentiating TLS formation as a promising approach to enhance anti-tumor immunity. We propose that lessons learned from BCG immunotherapy success could be applied to not only augment such microbe-based therapeutics but also lead to similar adjunctive IFN-1 activating approaches to improve response to immune checkpoint blockade therapy in cancer.

The skin epithelial layer acts as an important immunological barrier against pathogens and is capable of recognizing and responding to pathogen associated molecular patterns (PAMPs) in human and mouse models. Although presumed, it is unknown whether amphibian skin epithelial cells exhibit the ability to respond to PAMPs such as viral double-stranded RNA (dsRNA). To address this, two cell lines from the dorsal skin (Xela DS2) and ventral skin (Xela VS2) of the African clawed frog (Xenopus laevis) were established. Xela DS2 and Xela VS2 cells have an epithelial-like morphology, express genes associated with epithelial cells, and lack senescence-associated beta-galactosidase activity. Cells grow optimally in 70% Leibovitz’s L-15 medium supplemented with 15% fetal bovine serum at 26°C. Upon treatment with poly(I:C), a synthetic viral dsRNA analogue and known type I interferon inducer, Xela DS2 and Xela VS2 exhibit marked upregulation of key pro-inflammatory and antiviral transcripts suggesting frog epithelial cells participate in the recognition of extracellular viral dsRNA and production of local inflammatory signals; similar to human and mouse models. Currently, these are the only known Xenopus laevis skin epithelial-like cell lines and will be important for future research in amphibian epithelial cell biology, initial host-pathogen interactions, and rapid screening of the effects of environmental stressors, including contaminants, on frog skin epithelial cells.

For the last 50 years we have known of a broad-spectrum agent tilorone dihydrochloride (Tilorone). This is a small-molecule orally bioavailable drug that was originally discovered in the USA and is currently used clinically as an antiviral in Russia and the Ukraine. Over the years there have been numerous clinical and non-clinical reports of its broad spectrum of antiviral activity. More recently we have identified additional promising antiviral activities against Middle East Respiratory Syndrome, Chikungunya, Ebola and Marburg which highlights that this old drug may have other uses against new viruses. This may in turn inform the types of drugs that we need for virus outbreaks such as for the new coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Tilorone has been long neglected by the west in many respects but it deserves further reassessment in light of current and future needs for broad-spectrum antivirals.

BackgroundT-cell-engaging CD3-bispecific antibodies (CD3-bsAbs) are promising modalities for cancer immunotherapy. Although this therapy has reached clinical practice for hematological malignancies, the absence of sufficient infiltrating T cells is a major barrier for efficacy in solid tumors. In this study, we exploited oncolytic reovirus as a strategy to enhance the efficacy of CD3-bsAbs in immune-silent solid tumors.MethodsThe mutant p53 and K-ras induced murine pancreatic cancer model KPC3 resembles human pancreatic ductal adenocarcinomas with a desmoplastic tumor microenvironment, low T-cell density and resistance to immunotherapy. Immune-competent KPC3 tumor-bearing mice were intratumorally injected with reovirus type 3 Dearing strain and the reovirus-induced changes in the tumor microenvironment and spleen were analyzed over time by NanoString analysis, quantitative RT-PCR and multicolor flow cytometry. The efficacy of reovirus in combination with systemically injected CD3-bsAbs was evaluated in immune-competent mice with established KPC3 or B16.F10 tumors, and in the close-to-patient human epidermal growth factor receptor 2 (HER2)+ breast cancer model BT474 engrafted in immunocompromised mice with human T cells as effector cells.ResultsReplication-competent reovirus induced an early interferon signature, followed by a strong influx of natural killer cells and CD8+ T cells, at the cost of FoxP3+ Tregs. Viral replication declined after 7 days and was associated with a systemic activation of lymphocytes and the emergence of intratumoral reovirus-specific CD8+ T cells. Although tumor-infiltrating T cells were mostly reovirus-specific and not tumor-specific, they served as non-exhausted effector cells for the subsequently systemically administered CD3-bsAbs. Combination treatment of reovirus and CD3-bsAbs led to the regression of large, established KPC3, B16.F10 and BT474 tumors. Reovirus as a preconditioning regimen performed significantly better than simultaneous or early administration of CD3-bsAbs. This combination treatment induced regressions of distant lesions that were not injected with reovirus, and systemic administration of both reovirus and CD3-bsAbs also led to tumor control. This suggests that this therapy might also be effective for metastatic disease.ConclusionsOncolytic reovirus administration represents an effective strategy to induce a local interferon response and strong T-cell influx, thereby sensitizing the tumor microenvironment for subsequent CD3-bsAb therapy. This combination therapy warrants further investigation in patients with non-inflamed solid tumors.