Interferon System Research Articles

Endogenous ZAP is associated with altered Zika virus infection phenotype

The zinc finger antiviral protein 1 (ZAP) has broad antiviral activity. ZAP is an interferon (IFN)-stimulated gene, which itself may enhance type I IFN antiviral response. In a previous study, Zika virus was identified as ZAP-resistant and not sensitive to ZAP antiviral activity. Here, we found that ZAP was associated with the inhibition of Zika virus in Vero cells, in the absence of a robust type I IFN system because Vero cells are deficient for IFN-alpha and -beta. Also, quantitative RNA-seq data indicated that endogenous ZAP is associated with altered global gene expression both in the steady state and during Zika virus infection. Further studies are warranted to elucidate this IFN-alpha and -beta independent anti-Zika virus activity and involvement of ZAP.

Analysis of GCRV Pathogenesis and Therapeutic Measures Through Proteomic and Metabolomic Investigations in GCRV-Infected Tissues of Grass Carp (Ctenopharyngodon idella).

Hemorrhagic disease caused by grass carp reovirus (GCRV) infection is a major problem affecting the grass carp aquaculture industry. Therefore, inhibiting the spread of GCRV infection is of great economic significance. Herein, we sequenced five tissues (gill, liver, intestine, kidney, and muscle) from grass carp before and after GCRV infection using data-independent acquisition proteomic and untargeted metabolomic technologies, and quantitatively identified 10,808 proteins and 4040 metabolites. Then, we analyzed the differentially expressed proteins (DEPs) and metabolites (DEMs) before and after GCRV infection in the five tissues. Gene ontology analysis revealed that the five tissue DEPs were enriched in metabolic, including carbohydrate and lipid metabolic processes. Chemical taxonomy analysis showed that the categories of DEMs mainly included carbohydrates and lipids, such as fatty acids, glycerophospholipids, steroids, and their derivatives. Both the proteomic and the metabolomic data showed that GCRV affected the carbohydrate and lipid metabolism in the host. Shared pathway analysis was performed at both the protein and metabolic levels, showing significant enrichment of the glycolysis and pentose phosphate pathways (p < 0.001). Further analysis of glycolysis and pentose phosphate pathway inhibitors revealed that these two pathways are important for GCRV replication. As the kidney was the most affected among the five tissues, we analyzed the butanoate metabolism in the kidney, which revealed that most of the differentially expressed proteins and differently expressed metabolites in the butanoate metabolism were related to the TCA cycle. Further investigation showed that fumaric acid, an intermediate product in the TCA cycle, significantly inhibited GCRV replication in the CIK cells (p < 0.001), and that this inhibitory effect may be related to its induction of interferon system activation. The addition of fumaric acid to feed increased the survival rate of juvenile grass carp by 19.60% during GCRV infection, and protected the tissues of those infected with GCRV, making it a potential anti-GCRV feed additive. Our results provide new perspectives on GCRV pathogenesis and antiviral strategies for grass carp.

Interferon Regulatory Factors (IRF1, IRF4, IRF5, IRF7 and IRF9) in Sichuan taimen (Hucho bleekeri): Identification and Functional Characterization

Background/Objectives: Interferon regulatory factors (IRFs) are multifunctional transcription factors that play important roles in the transcriptional regulation of interferons and in the immune response to pathogens. Therefore, studying the interferon system in fish is highly relevant in the prevention and treatment of viral diseases. Methods: In this study, five IRF genes (IRF1, IRF4, IRF5, IRF7 and IRF9) were identified and characterized in Hucho bleekeri, and their expression profiles were determined after LPS and Poly(I:C) treatment. Results: These IRFs have typical DNA-binding domains and IRF-association domains. Amino acid sequence comparison revealed high homology between these IRFs and those of other vertebrates, with the highest homology being with other salmonid fish. Phylogenetic analysis revealed that these IRFs are divided into four subfamilies (IRF1, IRF3, IRF4 and IRF5), with both IRF4 and IRF9 belonging to the IRF4 subfamily. IRF genes were widely expressed in all of the tested tissues, with IRF1, IRF4 and IRF9 being highly expressed in the spleen and kidney and IRF5 and IRF7 highly expressed in the gonads. IRF1, IRF4 and IRF5 expression was induced at different time points post-LPS challenge. IRF7 and IRF9 expression in the spleen and head kidney was not significantly altered by LPS induction. Poly(I:C) treatment altered IRF expression more significantly than LPS treatment. Poly(I:C) significantly altered the spleen and head kidney expression of all five IRFs. Conclusions: These findings reveal the potential role of IRFs in the antiviral response of H. bleekeri and provide a reference for examining signal transduction pathways in the interferon system in fish.

Получение и оценка биологической активности экзогенной мРНК, кодирующей белок MxA человека

Human MxA protein induced by type I and III interferons is an important innate immunity mediator, it shows antiviral activity against a broad spectrum of RNA and DNA viruses. According to the latest data, the MxA protein overexpression increases chemotherapy sensitivity and represents one of the favorable prognostic factors in patients with breast cancer. The exogenous mRNA capable of intracellular MxA protein production not only has the potential for treatment of viral respiratory infection, but also can become an important fundamental research tool. The study aimed to construct and produce the exogenous mRNA encoding the functional human cytoplasmic MxA protein by in vitro transcription (IVT); to study its translational properties; to assess and identify the patterns of the expression of some interferon system genes in response to introduction of this exogenous mRNA into cells. As a result of the study, the exogenous mRNAs capable of effective translation (up to 20 ng/mL of protein from 100 ng of mRNA per well of the 96-well plate) in the eukaryotic cell systems were successfully constructed and produced by IVT (in the amount of up to 200 µg); diffuse distribution of the MxA protein in the MDCK cells was confirmed; significant changes in the expression of the interferon-stimulated genes, such as OAS1, PKR (EIF2AK2), MDA5, RIG-I, were revealed. Our further research will be focused on assessing the developed exogenous mRNAs’ therapeutic potential against influenza A and B viruses, respiratory syncytial virus, and coronavirus SARS-CoV-2.

Interferon-stimulated genes and their antiviral activity against SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) pandemic remains an international health problem caused by the recent emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of May 2024, SARS-CoV-2 has caused more than 775 million cases and over 7 million deaths globally. Despite current vaccination programs, infections are still rapidly increasing, mainly due to the appearance and spread of new variants, variations in immunization rates, and limitations of current vaccines in preventing transmission. This underscores the need for pan-variant antivirals and treatments. The interferon (IFN) system is a critical element of the innate immune response and serves as a frontline defense against viruses. It induces a generalized antiviral state by transiently upregulating hundreds of IFN-stimulated genes (ISGs). To gain a deeper comprehension of the innate immune response to SARS-CoV-2, its connection to COVID-19 pathogenesis, and the potential therapeutic implications, this review provides a detailed overview of fundamental aspects of the diverse ISGs identified for their antiviral properties against SARS-CoV-2. It emphasizes the importance of these proteins in controlling viral replication and spread. Furthermore, we explore methodological approaches for the identification of ISGs and conduct a comparative analysis with other viruses. Deciphering the roles of ISGs and their interactions with viral pathogens can help identify novel targets for antiviral therapies and enhance our preparedness to confront current and future viral threats.

Antiviral Mx proteins have an ancient origin and widespread distribution among eukaryotes.

First identified in mammals, Mx proteins are potent antivirals against a broad swathe of viruses. Mx proteins arose within the Dynamin superfamily of proteins (DSP), mediating critical cellular processes, such as endocytosis and mitochondrial, plastid, and peroxisomal dynamics. And yet, the evolutionary origins of Mx proteins are poorly understood. Using a series of phylogenomic analyses with stepwise increments in taxonomic coverage, we show that Mx proteins predate the interferon signaling system in vertebrates. Our analyses find an ancient monophyletic DSP lineage in eukaryotes that groups vertebrate and invertebrate Mx proteins with previously undescribed fungal MxF proteins, the relatively uncharacterized plant and algal Dynamin 4A/4C proteins, and representatives from several early-branching eukaryotic lineages. Thus, Mx-like proteins date back close to the origin of Eukarya. Our phylogenetic analyses also reveal that host-encoded and NCLDV (nucleocytoplasmic large DNA viruses)-encoded DSPs are interspersed in four distinct DSP lineages, indicating recurrent viral theft of host DSPs. Our analyses thus reveal an ancient history of viral and antiviral functions encoded by the Dynamin superfamily in eukaryotes.

Pleiotropicity of immunomodulating effects of synthetic thymic hexapeptide in chronic infectious and inflammatory diseases of the genital tract in immunocompromized women

A reason for a protracted course and frequent recurrence of pelvic inflammatory diseases (PID) is dysfunction of the immune system, which dictates the need of effective immunotherapeutic approaches. Objective: to clarify the features of immune system functioning, the interferon system, the profile of cytokines during the period of exacerbation of PID in women; to develop targeted immunotherapy using hexapeptide with assessment of its effectiveness. The studied immunocompromised women 20-40 years old with exacerbation of PID were divided into study groups (SG): SG1 (n = 22), SG2 (n = 70) before treatment; SG1a, SG1 receiving standard therapy; SG2a, SG2, receiving standard therapy and hexapeptide (HP, Imunofan®). The parameters of cellular and humoral immunity, levels of serum interferons, cytokines were determined. During exacerbation of PID, a decrease in IFNα was in both groups and IFNγ, more pronounced in SG1. In SG1, the levels of IL-6, TNFα were increased, the level of IL-8 was decreased. In SG2, the level of IL-6 was increased. An increase in IL-4, IL-10 was noted in both groups. Against the background of IFNα, IFNγ deficiency and cytokine imbalance, a significant decrease in T lymphocytes, T helper cells, B lymphocytes was observed in both study groups. After standard treatment, there was no recovery of IFNα, IFNγ. Only TNFα decreased, the levels of IL-1β, IL-6 were increased at the end of treatment, there was no increase in IL-4, IL-10 and restoration of cellular and humoral immunity. It leads to maintaining chronic inflammation and exacerbation of PID within 3 months after treatment. After immunotherapy including fHP, IFNα was restored in 50% cases. There was a decrease in IL-1β, TNFα without significant changes in the levels of IL-8, IL-18. Resolution of inflammation was accompanied by an increase in IL-10. The modulating effect of fHP on interferons and cytokines contributed to the restoration of T cell and humoral immunity, which is related to the onset of clinical remission of PID at an earlier time and an increase in the duration of the inter-relapse period. The positive effectiveness of immunotherapy with Imunofan® is due to its pleiotropic effects, which provides significant advantages compared to the use of standard therapy only.

Comparative assessment of interferon activity in influenza and COVID-19

Among respiratory viruses, the most serious complications are caused by influenza A and B viruses, as well as coronaviruses. Most studies determined the absolute content of interferons (IFNs) of different types in blood serum. However, serum IFN protein concentrations do not always reflect the level of antiviral protection. The purpose of this study was a comparative assessment of interferon status in patients with ARVI: influenza and the acute stage of COVID-19. Materials and methods. We used biomaterial in the form of whole blood samples from 113 patients with influenza and 110 patients in the acute phase of moderate COVID-19. The body’s antiviral defense during ARVI was assessed by determining the activity of type I and II interferons produced by blood leukocytes using the “Interferon status” method in a cell-virus system simulated in vitro. Results. This work reveals a statistically significant decrease in the biological activity of interferons produced by blood leukocytes in influenza and a deficiency of IFN activity in COVID-19, compared with reference values, and also shows possible prospects for the treatment of these nosologies with such immunoactive drugs as IFN inducers (cycloferon, Kagocel) and immunomodulators (ingavirin, multicomponent vaccine Immunovac-VP-4). Conclusion. The results of IFN activity are necessary to assess the antiviral potential of the body, especially with COVID-19, given the “novelty” of the infection, the severity and variety of its clinical manifestations. Today it is known that the SARS-CoV-2 virus is capable of penetrating not only into the epithelial cells of the upper respiratory tract, epithelial cells of the stomach and intestines, but also into the cells of the esophagus, heart, adrenal glands, bladder, brain, as well as into the vascular endothelium and macrophages. Coronavirus SARS-CoV-2 inhibits the expression of cellular genes, including innate immune genes, and has a negative effect on the IFN system. The use of IFN inducers and immunomodulators for influenza and COVID-19 has shown immunological feasibility and clinical promise.

Personalized approach to immune system rehabilitation in patients with year-round allergic rhinitis suffering from recurrent ARVI and recurrent herpes virus infections

Introduction. The problem of treating patients with respiratory allergopathology associated with recurrent infectious diseases, such as rARVI, frequent exacerbations of rCHVI, is becoming urgent. The presence of persistent inflammation and co-infection significantly complicates the treatment of immunocompromised patients with year-round allergic rhinitis (YAR) and necessitates the development of personalized programs, with the inclusion of immunomodulatory agents for restoring disorders in the immune system (IS) and interferon system (IFN). Materials and methods. The study group (SG) included 65 patients of both sexes aged 23–60 years, suffering from YAR associated with rARVI and rCHVI. The comparison group (CG) consisted of 50 healthy individuals, comparable in sex and age. All patients underwent standard physical, immunological and allergological examinations, including the use of serological (ELISA, ImmunoCUP), molecular genetic (PCR-RV) methods, FC, etc. Voluntary informed consent was obtained from all patients. The StatPlus computer program was used for statistical analysis. Results and discussion. In patients with YAR associated with rARVI and rCHVI, two variants of disorders in IS have been established, which are defined as pathological immunophenotypes (PIF). The PIF1 is characterized by a deficiency in the induced production of IFNα, a decrease in CD3⁻CD16⁺CD56+ EKKs, and a decrease in neutrophil granulocytes (NG). In the PIF2, along with a shortage of IFNα production, a decrease in EKKs is observed in combination with a decrease in CTLs, as well as a decrease in NGs. The clinical criteria for immunocompromising in this cohort of patients were studied. The severity of YAR symptoms was assessed by VAS. In order to correct the identified disorders in the IS and IFN system, complex personified IFN- and immunotherapy programs have been developed for each PIF, including prolonged local and systemic therapy with rIFNα2b in combination with antioxidants, as well as GMDP for patients of SG1, and for patients of SG2 — GMDP and hexapeptide. The high immunological and clinical efficacy of IFN- and immunotherapy was shown, which was expressed in the tendency to restore existing disorders, as well as in achieving control over the symptoms of YAR, with the possibility of reducing the volume of basic antiallergic pharmacotherapy.

Update on the pathophysiology and treatment of primary Sjögren syndrome.

Sjögren syndrome or Sjögren disease is a chronic form of autoimmune epithelitis characterized by lymphocytic infiltration of the exocrine glands, particularly the salivary and lacrimal glands, leading to progressive glandular dysfunction and subsequent xerostomia and xerophthalmia. Other common manifestations include pain and fatigue, various systemic manifestations and non-Hodgkin's lymphoma. Sjögren syndrome is therefore a complex and disabling disease associated with a reduced quality of life and with considerable long-term damage. Most of the available treatments are merely symptomatic with limited efficacy in both preventing glandular damage and suppressing systemic disease activity. In the past 10 years, great progress has been made in understanding the pathophysiology of Sjögren syndrome, opening new avenues towards a more targeted and individualized therapeutic approach to the disease. Indeed, several randomized controlled trials have just been completed or are poised to commence evaluating the effectiveness of novel drugs targeting both innate and adaptive immune pathways, including pro-inflammatory cytokines, the type I interferon system, B cell activation, B cell and T cell co-stimulation pathway, and ectopic germinal centre formation. Novel clinical trials are also ongoing exploring various targeted approaches (that is, IgG recycling inhibition, nuclease therapy and CAR-T cell therapy) for Sjögren syndrome.

Genetic polymorphisms in genes involved in the type I interferon system (STAT4 and IRF5): association with Asian SLE patients.

Systemic lupus erythematosus (SLE) is a common autoimmune disease with a polymorphic clinical presentation involving multisystem damages with significant differences in prevalence and disease severity among different ethnic groups. Although genetic, hormonal, and environmental factors have been demonstrated to contribute a lot to SLE, the pathogenesis of SLE is still unknown. Numerous evidence revealed that gene variants within the type I interferons (IFN) signaling pathway performed the great genetic associations with autoimmune diseases including SLE. To date, through genome-wide association studies (GWAS), genetic association studies showed that more than 100 susceptibility genes have been linked to the pathogenesis of SLE, among which TYK2, STAT1, STAT4, and IRF5 are important molecules directly connected to the type I interferon signaling system. The review summarized the genetic associations and the detailed risk loci of STAT4 and IRF5 with Asian SLE patients, explored the genotype distributions associated with the main clinical manifestations of SLE, and sorted out the potential reasons for the differences in susceptibility in Asia and Europe. Moreover, the therapies targeting STAT4 and IRF5 were also evaluated in order to propose more personalized and targeted treatment plans in SLE.

The interferon-induced protein, IFIT2, requires RNA-binding activity and neuronal expression to protect mice from intranasal vesicular stomatitis virus infection.

The interferon (IFN) system protects mammals from diseases caused by virus infections. IFN synthesis is induced by pattern recognition receptor signaling pathways activated by virus infection. IFN is secreted from the infected cells and acts upon neighboring cells by binding cell surface receptors and triggering induction of hundreds of IFN-stimulated genes and proteins, many of which block different steps of virus replication. The IFN-induced tetratricopeptide repeat proteins (IFIT) are a family of RNA-binding proteins. We and others have previously reported that IFIT2 protects mice from many neurotropic RNA viruses; indeed, Ifit2-/- mice are very susceptible to intranasal or subcutaneous infections with vesicular stomatitis virus (VSV). Here, using a newly generated conditional knockout mouse, we report that ablation of Ifit2 expression only in neuronal cells was sufficient to render mice susceptible to neuropathogenesis caused by intranasal, but not subcutaneous, infection of VSV. Another genetically modified mouse line, expressing a mutant IFIT2 that cannot bind RNA, was as susceptible to VSV infection as Ifit2-/- mice. These results demonstrated that IFIT2 RNA-binding activity is essential for protecting mice against neurological diseases caused by intranasal infection of VSV.IMPORTANCEInterferon's (IFN's) antiviral effects are mediated by the proteins encoded by the interferon-stimulated genes. IFN-stimulated genes (IFIT2) is one such protein, which inhibits replication of many RNA viruses in the mouse brain and the resultant neuropathology. Our study sheds light on how IFIT2 works. By ablating Ifit2 expression only in neuronal cells, using a newly generated conditional knockout mouse line, we showed that Ifit2 induction in the neurons of the infected mouse was necessary for antiviral function of interferon. IFIT2 has no known enzyme activity; instead, it functions by binding to cellular or viral proteins or RNAs. We engineered a new mouse line that expressed a mutant IFIT2 that cannot bind RNA. These mice were very susceptible to infection with vesicular stomatitis virus indicating that the RNA-binding property of IFIT2 was essential for its antiviral function in vivo.

Interferon-Stimulated Genes that Target Retrovirus Translation.

The innate immune system, particularly the interferon (IFN) system, constitutes the initial line of defense against viral infections. IFN signaling induces the expression of interferon-stimulated genes (ISGs), and their products frequently restrict viral infection. Retroviruses like the human immunodeficiency viruses and the human T-lymphotropic viruses cause severe human diseases and are targeted by ISG-encoded proteins. Here, we discuss ISGs that inhibit the translation of retroviral mRNAs and thereby retrovirus propagation. The Schlafen proteins degrade cellular tRNAs and rRNAs needed for translation. Zinc Finger Antiviral Protein and RNA-activated protein kinase inhibit translation initiation factors, and Shiftless suppresses translation recoding essential for the expression of retroviral enzymes. We outline common mechanisms that underlie the antiviral activity of multifunctional ISGs and discuss potential antiretroviral therapeutic approaches based on the mode of action of these ISGs.

Clinical Trials of Interferon Inhibitors in Systemic Lupus Erythematosus and Preliminary Real-World Efficacy of Anifrolumab.

Approval of anifrolumab for the treatment of moderate-to-severe systemic lupus erythematosus (SLE) in 2021 marked the success of a long quest to target the interferon system, in a disease wherein the latter has long been considered to play a pivotal role. Prior to anifrolumab, a number of agents had been tested in early phase clinical trials in patients with SLE, with equivocal results. Following its approval and marketing in several countries, the first reports regarding efficacy and safety in real-life clinical settings have been published, which suggest remarkable efficacy in skin manifestations of the disease, even after prior failure to multiple immunosuppressive therapies. In this report, we provide a short overview of IFN inhibitors that have been used in clinical trials of SLE, with a focus on anifrolumab; we also review all available evidence to date regarding its real-world efficacy and safety.

Current Status of Vaccines for Porcine Reproductive and Respiratory Syndrome: Interferon Response, Immunological Overview, and Future Prospects.

Porcine reproductive and respiratory syndrome (PRRS) remains a formidable challenge for the global pig industry. Caused by PRRS virus (PRRSV), this disease primarily affects porcine reproductive and respiratory systems, undermining effective host interferon and other immune responses, resulting in vaccine ineffectiveness. In the absence of specific antiviral treatments for PRRSV, vaccines play a crucial role in managing the disease. The current market features a range of vaccine technologies, including live, inactivated, subunit, DNA, and vector vaccines, but only modified live virus (MLV) and killed virus (KV) vaccines are commercially available for PRRS control. Live vaccines are promoted for their enhanced protective effectiveness, although their ability to provide cross-protection is modest. On the other hand, inactivated vaccines are emphasized for their safety profile but are limited in their protective efficacy. This review updates the current knowledge on PRRS vaccines' interactions with the host interferon system, and other immunological aspects, to assess their current status and evaluate advents in PRRSV vaccine development. It presents the strengths and weaknesses of both live attenuated and inactivated vaccines in the prevention and management of PRRS, aiming to inspire the development of innovative strategies and technologies for the next generation of PRRS vaccines.

The impact of functional feed on Atlantic salmon (Salmo salar) systemic immune response to high and low levels of sea lice infection (Lepeophtheirus salmonis) and co-infection with infectious salmon anemia virus

Due to the nature of open-pen farming, salmon are exposed to numerous pathogens shared by other farms and their wild counterparts. Industry must manage these outbreaks through vaccination, oral or bath treatments, and more recently through functional feed administration. Globally, the most important pathogens of salmon are sea lice (Lepeophtheirus salmonis, and Caligus rogercresseyi) not only due to their direct impacts on the host, but indirectly by enhancing host susceptibility to co-infection. This study aims to characterize molecular responses during a co-infection of L. salmonis and a salmon orthomyxovirus (infectious salmon anemia virus; ISAv) under administration of four functional feed diets: a control feed with a low 0.3 % EPA/DHA + high-ω6 (Ctrl), an EPA/DHA enriched pro-inflammatory diet 1 % EPA/DHA + high-ω6; FA + I), an EPA/DHA enriched anti-inflammatory diet (1 % EPA/DHA+high-ω3; FA-I), and a low EPA/DHA feed (0.3 %) with an immunostimulant added (0.3 % EPA/DHA + high-ω6 + immunostimulant (IS); Ctrl + IS). Atlantic salmon (40 fish per tank; 8 tanks per feed) were acclimated to one of the four experimental diets. Sea lice copepodids were introduced to all experimental tanks and 10 fish sampled from each tank at each time point (prior to infection [−3], and 11, 33, 47 days post infection). A high virulence ISAv isolate (ISAV-HPR4) was intraperitoneally injected into donor fish 6 days prior to their transfer into the experimental tanks for cohabitation (ca. 10–15 % of tank density; 4 tanks per feed group) to achieve peak shedding rates at time of stocking. Fatty acid enriched (FA + I and FA-I) diets had a significant impact on sea lice abundance on fish infected with lice only. Gene expression profiles measured by reverse transcriptase-qPCR showed significant upregulation in several antiviral genes (irf7b and mxb) associated with the interferon system in all but the fish fed the immunostimulating diet. An increase in transcript levels (irf7b, isg15a, mmp-9, mxb) accounted for high lice and high viral loads in diets FA + I and Ctrl + IS. As feeds successful at reducing lice were the least successful in survival of co-infected fish, there appears to be a trade-off for better anti-parasitic responses, which is enhanced through dietary supplementation. This will require further research to ensure careful selection of feed combinations and expected co-infection challenges.

Metapneumovirus infection in children in modern conditions: clinical and immunological parallels

Aim. The study of metapneumovirus infection clinical and immunological features in children of different ages. Patients and methods. An analysis of 22 patients with mono-metapneumovirus infection is presented. There were 7 (31.8%) children under the age of 1 year, 8 (36.4%) — 1—3 years old, 7 (31.8%) — 4—7 years old. Diagnosis of metapneumovirus infection was based on clinical data and on PCR analysis of nasopharyngeal mucus. Immunological studies in 10 patients included immunophenotype of patient's blood lymphocytes and analysis of the interferon system. Results. Among children under the age of 1 year 71.4 ± 17.1% were diagnosed with obstructive bronchitis and 28.6 ± 17.1% had pneumonia. In patients aged 1—3 years pneumonia was observed more often (55.6 ± 17.6%), obstructive bronchitis — less often (33.3 ± 16.7%), and in 11.1 ± 11.1% bronchitis occurred without symptoms of bronchial obstruction. In children aged 4—7 years only the upper respiratory tract was affected in 14.3 ± 13.2% of cases, 57.1 ± 18.7% developed bronchitis and 28.6 ± 17.1% — pneumonia. The immune system state was characterized by a low content of T-helpers (CD3 + CD4 + ) and NK-cells (CD3-CD16 + CD56 + ) detected in 7 (70%) and 5 (50%) patients respectively. IFNγ production was reduced in 70% of children and IFNα in 80%. Conclusion. Metapneumovirus infection in children in modern conditions is characterized by the frequent development of inflammatory changes in the bronchopulmonary system in the early stages of the disease. The disease severity was due to both manifestations of respiratory failure and severe symptoms of intoxication. Immunological studies revealed changes in the cellular link and the interferon system in all age groups.

Abstract 3927: Metabolic inhibition of BATF2 dampens type-I interferon-mediated immune sensing of cancer

Abstract Background: The type-I interferon (IFN-I) system appeared before adaptive immunity during evolution as the primary defense system for metazoans. Initiating cancers frequently disable this first line of defense before establishing T-cell exhaustion programs. However, the molecules linking IFN-I, danger signals, and unique metabolic conditions in the tumor microenvironment (TME) remain to be discovered. Methods: We performed centered log-ratio transformation on the cluster composition data using the R-package ALDEx2 to compare the clusters in single-cell data sets. IFN-I signatures were quantified using qPCR and ELISA. ChIP was performed to examine the H3K27me3 epigenetic marker on the promoters of BATF2 and IFN-I genes in bone marrow-derived macrophage (BMDM). For the cigarette smoke carcinogen-induced head and neck squamous cell carcinoma (HNSCC) model, experimental mice were fed with water containing 50 μg/mL of 4-NQO for 16 weeks and then switched to regular feeding water. We started monitoring 10 weeks post-treatment for the number, location, and diameter of the lesions. TME analysis was performed using single-cell RNA-Seq and multispectral imaging. Results: We analyzed three large independent cohorts of patients and found that BATF2 strongly correlated with the immunogenicity of HNSCC, with a stronger correlation coefficient than STING. We generated Batf2−/− BMDM and found that Batf2 deficiency rendered cells insensitive to STING agonists. However, the dsRNA-induced IFN-I activation was not affected by Batf2 deletion. BATF2 interacted with STING, promoting its phosphorylation and oligomerization. Glutamine is enriched in the TME. We found that glutamine inhibited the BATF2-STING pathway by increasing a repressive epigenetic marker, H3K27me3, on the promoters of BATF2 and IFNB1. The expression of BATF2 in cancer cells resulted in IFN-I- and γδ T-cell-dependent tumor rejection. Single-cell immune profiling showed that Batf2-expressing tumors contained significantly expanded Cxcl10+ myeloid cells and γδ T-cells. Utilizing the 4-NQO-induced HNSCC model, we found that BATF2 limited suppressive myeloid cells and expanded effector cells in the TME, serving as a potent tumor suppressor. Conclusion: BATF2 is a glutamine-responsive tumor suppressor that limits tumor initiation by engaging the STING-γδ T-cell pathway. Acknowledgments: This work was supported by NIH grants R01 DE026728, U01 DE029255, R01 DE031951, and U01 DE033330 Citation Format: Wang Gong, Hülya Taner, Yuesong Wu, Zaiye Li, Wanqing Cheng, Christopher Donnelly, Felipe Nör, Yumin He, Zackary Fitzsimonds, Jianqian Li, Haitao Wen, Steven Chinn, Yuying Xie, James Moon, Yu Leo Lei. Metabolic inhibition of BATF2 dampens type-I interferon-mediated immune sensing of cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3927.

Revealing the changes in signaling pathways caused by tofacitinib in patients with rheumatoid arthritis through RNA sequencing and the correlation with clinical parameters.

The current study is to accelerate the understanding of how tofacitinib works in patients with rheumatoid arthritis (RA) due to the lack of relevant information. We selected ten patients with active RA and obtained the expression profile for their peripheral blood mononuclear cells before and after the tofacitinib treatment by RNA sequencing. The gene set enrichment analysis was conducted, and the significantly enriched gene sets were identified. The hub gene highly correlated with clinical parameters in the gene set was selected. We constructed the weighted gene co-expression network, linked modules with clinical indicators, and screened hub genes. The expression of representative hub genes was validated by real-time quantitative PCR (qPCR). Gene set interferon (IFN) α and IFN β signaling was the most significantly down-regulated after tofacitinib treatment. In this gene set, genes Oas2 and Oasl showed a significant positive correlation with morning stiffness. In co-expression network, gene Vgll3 from the violet module with the highest correlation coefficient, was positively correlated with morning stiffness. Among them, Oasl and Vgll3 have shown significant down-regulation in qPCR validation. Our results highlighted the role of type I IFN, mainly including IFN α and IFN β, in the pathogenesis of RA and action for tofacitinib, and provided a new entry point for further elucidating the mechanism of morning stiffness. Key Points • Gene set IFN α and IFN β signaling was the most significantly down-regulated after tofacitinib treatment in RA patients. • Gene Oasl and Vgll3 were correlated with morning stiffness and significantly down-regulated due to the action of tofacitinib. • Type I IFN system was highlighted in the action of tofacitinib.

Low CD4 + T cell count is related to specific anti-nuclear antibodies, IFNα protein positivity and disease activity in systemic lupus erythematosus pregnancy

BackgroundLymphopenia, autoantibodies and activation of the type I interferon (IFN) system are common features in systemic lupus erythematosus (SLE). We speculate whether lymphocyte subset counts are affected by pregnancy and if they relate to autoantibody profiles and/or IFNα protein in SLE pregnancy.MethodsRepeated blood samples were collected during pregnancy from 80 women with SLE and 51 healthy controls (HC). Late postpartum samples were obtained from 19 of the women with SLE. Counts of CD4 + and CD8 + T cells, B cells and NK cells were measured by flow cytometry. Positivity for anti-nuclear antibodies (ANA) fine specificities (double-stranded DNA [dsDNA], Smith [Sm], ribonucleoprotein [RNP], chromatin, Sjögren’s syndrome antigen A [SSA] and B [SSB]) and anti-phospholipid antibodies (cardiolipin [CL] and β2 glycoprotein I [β2GPI]) was assessed with multiplexed bead assay. IFNα protein concentration was quantified with Single molecule array (Simoa) immune assay. Clinical data were retrieved from medical records.ResultsWomen with SLE had lower counts of all lymphocyte subsets compared to HC throughout pregnancy, but counts did not differ during pregnancy compared to postpartum. Principal component analysis revealed that low lymphocyte subset counts differentially related to autoantibody profiles, cluster one (anti-dsDNA/anti-Sm/anti-RNP/anti-Sm/RNP/anti-chromatin), cluster two (anti-SSA/anti-SSB) and cluster three (anti-CL/anti-β2GPI), IFNα protein levels and disease activity. CD4 + T cell counts were lower in women positive to all ANA fine specificities in cluster one compared to those who were negative, and B cell numbers were lower in women positive for anti-dsDNA and anti-Sm compared to negative women. Moreover, CD4 + T cell and B cell counts were lower in women with moderate/high compared to no/low disease activity, and CD4 + T cell count was lower in IFNα protein positive relative to negative women. Finally, CD4 + T cell count was unrelated to treatment.ConclusionLymphocyte subset counts are lower in SLE compared to healthy pregnancies, which seems to be a feature of the disease per se and not affected by pregnancy. Our results also indicate that low lymphocyte subset counts relate differentially to autoantibody profiles, IFNα protein levels and disease activity, which could be due to divergent disease pathways.