Method to Assess Immunosenescent CD8+ T Cells in Respiratory Viral Infections.

Immunosenescence is a well-characterized phenomenon that occurs with increasing age in all immune and somatic cells. In order to best study immunosenescence, it is imperative to develop methods to accurately identify immunosenescent cells. Elderly patients are known to have impaired immune responses to respiratory viruses, and it is hypothesized that this is due, in part, to immunosenescent, terminally exhausted CD8+ T cells. To test this hypothesis, we developed an aged mouse model and a flow cytometry protocol using the Cytek® Aurora to assess the CD8+ T-cell response during respiratory viral infection and identify immunosenescent CD8+ T cells. This protocol and our aged mouse model have great potential to be incredibly valuable for future studies elucidating how to rejuvenate and possibly reverse immunosenescent CD8+ T cells, which could improve the immune response to respiratory viruses in this at-risk population.

The Application of Organoids in the Study of Antiviral Innate Immunity.

Antiviral innate immunity plays a critical role in the defense against viral infections, yet its complex interactions with viruses have been challenging to study using traditional models. Organoids, three-dimensional (3D) tissue-like structures derived from stem cells, have emerged as powerful tools for modeling human tissues and studying the complex interactions between viruses and the host innate immune system. This chapter summarizes relevant applications of organoids in antiviral innate immunity studies and provides detailed information and experimental procedures for using organoids to study antiviral innate immunity.

Zebrafish as a Model for Investigating Antiviral Innate Immunity.

Zebrafish is a widely used model organism in genetics, developmental biology, pathology, and immunology research. Due to their fast reproduction, large numbers, transparent early embryos, and high genetic conservation with the human genome, zebrafish have been used as a model for studying human and fish viral diseases. In particular, the ability to easily perform forward and reverse genetics and lacking a functional adaptive immune response during the early period of development establish the zebrafish as a favored option to assess the functional implication of specific genes in the antiviral innate immune response and the pathogenesis of viral diseases. In this chapter, we detail protocols for the antiviral innate immunity analysis using the zebrafish model, including the generation of gene-overexpression zebrafish, generation of gene-knockout zebrafish by clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology, methods of viral infection in zebrafish larvae, analyzing the expression of antiviral genes in zebrafish larvae using qRT-PCR, Western blotting and transcriptome sequencing, and in vivo antiviral assays. These experimental protocols provide effective references for studying the antiviral immune response in the zebrafish model.

Fatal Hyperacute Liver Failure due to Varicella Zoster Virus Immediately After Living-Donor Liver Transplantation: A Case Report and Review of the Literature.

BACKGROUND: Although acute hepatitis caused by varicella zoster virus mostly develops in immunocompromised patients, hyperacute liver failure is very rare. To our knowledge, there are no previous reports on liver transplant patients. METHODS: We report the first case of fatal hyperacute liver failure due to varicella zoster virus immediately after living-donor liver transplantation without cutaneous lesions and review the literature. RESULT: The present case exhibited rapid development and progression of acute liver failure from postoperative days 11-13, despite being seropositive for varicella zoster virus but unvaccinated and on immunosuppression before transplantation. Especially in solid organ transplantation, only six cases of severe acute liver failure that included hepatic encephalopathy and/or impaired consciousness and sudden extremely high (> 4000 U/L) serum aspartate aminotransferase levels have been reported in heart, lung, and kidney transplant patients. CONCLUSIONS: Early diagnosis of hyperacute liver failure due to varicella zoster virus is challenging because the disease progresses rapidly and skin lesions are absent.

EPS T14 from Bacillus licheniformis Prevents Infection of Human Nasal Epithelial Cells by Respiratory Viruses.

Background: Respiratory viral infections are a leading cause of severe diseases and mortality; therefore, novel treatments effective for their prevention are highly requested. Here, we identified a broad-spectrum antiviral activity of a natural exopolysaccharide, EPS T14, purified from a marine thermotolerant strain of Bacillus licheniformis strain T14. Methods: The effects on human normal nasal epithelial cells (HNEpCs) following treatment with EPS T14 was evaluated at different time points and with increasing concentration of compound. To assess the antiviral properties, viability of HNEpCs treated with EPS T14 was analysed following infection with different respiratory viruses. Results: Neither toxicity nor pro-inflammatory properties were observed in vitro on HNEpCs treated with EPS T14 up to high concentrations, thus ensuring its safety. Cell culture-based assays revealed that treatment of HNEpCs with EPS T14 (used at 400ug/mL) results in efficient prevention of cell infection by different respiratory viruses through physically hindering the entry of the viruses via cell surface receptors. Interestingly, in addition to this prophylactic antiviral activity, EPS T14 also shows a long-lasting efficacy by inhibiting viral spread in the cell culture. Finally, combination of EPS T14 with a hypertonic saline solution shows a synergistic antiviral activity. Conclusion: EPS T14 can exert both prophylactic and therapeutic antiviral activity by blocking viral attachment to cellular receptors and could therefore represent a promising antiviral agent for preventing infections by different respiratory viruses.

Unmasking bocavirus: is it a co-infectious agent or an actual respiratory pathogen?

OBJECTIVES: Although human bocavirus (HBoV) is primarily linked to respiratory tract infections, its exact role as a respiratory pathogen remains unclear. This study aims to investigate HBoV detection rates, as well as clinical, laboratory, microbiological, and radiological characteristics, length of stay in the emergency department (ED), rate of hospitalization, and severity of illness in cases where HBoV is detected in respiratory secretions. METHODS: We conducted a retrospective analysis of all consecutive patients under 18 years who visited a large-volume tertiary pediatric ED from January to December 2023 and tested positive for HBoV in their respiratory viral panel (RVP). RESULTS: Among the 14,315 patients who underwent RVP testing during the study period, 591 (4%) tested positive for HBoV. After excluding those with incomplete data, 528 patients (57% male) were included in the analyses. The median age was 2.8 [1.2-4.9] years. The most common symptoms were cough (67%), fever (58%), runny nose/nasal congestion/sore throat (34%), and respiratory distress (24%). Thirty percent of the patients had a history of antibiotic use before admission. Thirteen percent of the patients had at least one chronic illness. Co-infection with HBoV occurred in 37% of the patients, with respiratory syncytial virus (RSV) being the most frequently co-detected virus (45%). Lymphopenia was documented in 12% of patients, and 36% had elevated C-reactive protein levels (median 21 [12-38] g/dl). Abnormal chest X-rays were noted in 85% of patients. The management approach included outpatient care for more than half of the patients (69%). Clinical severity was classified as high in 11% of patients (n = 60), necessitating ICU admission. CONCLUSION: Although typically mild, HBoV infections can escalate to severe respiratory illnesses, requiring respiratory support and intensive care.

Viral Infections and the Glutathione Peroxidase Family: Mechanisms of Disease Development.

Significance: The glutathione peroxidase (GPx) family is recognized for its essential function in maintaining cellular redox balance and countering the overproduction of reactive oxygen species (ROS), a process intricately linked to the progression of various diseases including those spurred by viral infections. The modulation of GPx activity by viruses presents a critical juncture in disease pathogenesis, influencing cellular responses and the trajectory of infection-induced diseases. Recent Advances: Cutting-edge research has unveiled the GPx family's dynamic role in modulating viral pathogenesis. Notably, GPX4's pivotal function in regulating ferroptosis presents a novel avenue for the antiviral therapy. The discovery that selenium, an essential micronutrient for GPx activity, possesses antiviral properties has propelled us toward rethinking traditional treatment modalities. Critical Issues: Deciphering the intricate relationship between viral infections and GPx family members is paramount. Viral invasion can precipitate significant alterations in GPx function, influencing disease outcomes. The multifaceted nature of GPx activity during viral infections suggests that a deeper understanding of these interactions could yield novel insights into disease mechanisms, diagnostics, prognostics, and even chemotherapeutic resistance. Future Directions: This review aims to synthesize current knowledge on the impact of viral infections on GPx activity and expression and identify key advances. By elucidating the mechanisms through which GPx family members intersect with viral pathogenesis, we propose to uncover innovative therapeutic strategies that leverage the antioxidant properties of GPx to combat viral infections. The exploration of GPx as a therapeutic target and biomarker holds promise for the development of next-generation antiviral therapies. Antioxid. Redox Signal. 00, 000-000.

Genotypic diversity and immunological implications of porcine circovirus: Inspiration from PCV1 to PCV4.

Porcine circovirus (PCV) is a group of DNA viruses that cause diseases in pigs, with multiple genotypes ranging from PCV1 to PCV4. PCV1 is generally considered non-pathogenic, while PCV2 can cause severe immune system damage, especially associated with porcine multisystemic wasting syndrome (PMWS). PCV2 has a genetic homology of about 68 % but differs from PCV1 in antigenicity and phenotype. PCV3 and PCV4 have lower genetic homology with PCV1 and PCV2, with limited research available on their pathogenicity. During virus infection, the host's innate immune system detects PCVs through pattern recognition receptors (PRRs) like TLRs and NLRs. PCV disrupts immune pathways, including interferon and NF-κB pathways, aiding viral replication and causing immunosuppression. This review systematically compares the characteristics and pathogenicity of different genotypes of PCV and their interactions with the host's immune system, aiming to better understand the mechanisms of PCV infection and provide a theoretical basis for prevention and treatment.

Protective Effect of Allergen Immunotherapy in Patients With Allergic Rhinitis and Asthma Against COVID-19 Infection: Observational, Nationwide, and Multicenter Study.

Background: Allergic diseases are associated with an increased susceptibility to respiratory tract infections. Although allergen immunotherapy (AIT) alters the course of allergies, there is limited evidence from clinical practice demonstrating its ability to enhance the host defense against pathogens. Objective: The aim of this study was to investigate the protective effect of AIT against viral infection in patients with allergic rhinitis (AR) and allergic asthma (AS) based on clinical evidence. Methods: A multicenter, questionnaire-based survey was conducted during a tremendous surge in COVID-19 cases between February 10, 2023, and March 15, 2023, in 81 centers across China recruiting healthy volunteers and patients with AR and AS to investigate the clinical outcomes of COVID-19 infection. Results: Of 10,151 participants recruited in the survey, 3654 patients and 2192 healthy volunteers who tested positive for COVID-19 were included in this analysis after screening. Overall, no significant differences in COVID-19 outcomes were observed between patients and healthy volunteers. An additional 451 patients were excluded due to their use of biologics as the sole add-on treatment, leaving 3203 patients in the further analysis. Of them, 1752 were undergoing routine medication treatment (RMT; the RMT group), whereas 1057 and 394 were receiving AIT and a combination of AIT and omalizumab (OMA) as adjunct therapies to RMT, respectively (AIT+RMT and AIT+OMA+RMT groups). The AIT group showed milder COVID-19 symptoms, shorter recovery periods, and a lower likelihood of hospitalization or emergency department visits than the RMT group (all P<.05). After adjusting for confounding factors, including demographic characteristics and COVID-19 vaccination, AIT remained a significant protective factor associated with shorter recovery time (adjusted odds ratio [OR] 0.62, 95% CI 0.52-0.75; adjusted P<.001) and a lower incidence of hospitalization or emergency department visits (adjusted OR 0.73, 95% CI 0.54-0.98; adjusted P=.03). Furthermore, the AIT+OMA+RMT group showed greater protection with a shorter recovery time (adjusted OR 0.51, 95% CI 0.34-0.74; adjusted P<.001) than the AIT+RMT group. Conclusions: Our multicenter observational study provides valuable clinical evidence supporting the protective effect of AIT against COVID-19 infection in patients with AR and AS.

Unveiling the promise: Exosomes as game-changers in anti-infective therapy.

Extracellular vesicles (EVs)-based intercellular communication (through exosomes, microvesicles, and apoptotic bodies) is conserved across all kingdoms of life. In recent years, exosomes have gained much attention for targeted pharmaceutical administration due to their unique features, nanoscale size, and capacity to significantly contribute to cellular communication. As drug delivery vehicles, exosomes have several advantages over alternative nanoparticulate drug delivery technologies. A key advantage lies in their comparable makeup to the body's cells, which makes them non-immunogenic. However, exosomes vesicles face several challenges, including a lack of an effective and standard production technique, decreased drug loading capacity, limited characterization techniques, and underdeveloped isolation and purification procedures. Exosomes are well known for their long-term safety and natural ability to transport intercellular nucleic acids and medicinal compounds across the blood-brain-barrier (BBB). Therefore, in addition to revealing new insights into exosomes' distinctiveness, the growing availability of new analytical tools may drive the development of next-generation synthetic systems. Herein, light is shed on exosomes as drug delivery vehicles in anti-infective therapy by reviewing the literature on primary articles published between 2002 and 2023. Additionally, the benefits and limitations of employing exosomes as vehicles for therapeutic drug delivery are also discussed.

Inhaled GM-CSF administered during ongoing pneumovirus infection alters myeloid and CD8 T cell immunity without affecting disease outcome.

Granulocyte-macrophage colony stimulating factor (GM-CSF) is a pleiotropic cytokine, able to promote both myelopoiesis and activation of immune cells. Particularly in the lung, GM-CSF plays an important homeostatic role in the development and maintenance of alveolar macrophages, and is therefore considered to play a role in respiratory virus infections such as influenza and SARS-CoV-2, although the benefits of GM-CSF treatment in clinical studies remain inconclusive. To address this, we tested inhaled GM-CSF treatment in the Pneumonia Virus of Mice (PVM) mouse model. Our findings show that local GM-CSF therapy during PVM disease increased local neutrophilia and monocyte-derived cell influx, but diminished CD8+ T cells responses. Despite this, the observed effects on T cells and myeloid cells did not result in an altered clinical outcome during PVM infection. We conclude that inhaled GM-CSF therapy cannot be considered as a universal protective therapy in respiratory virus infections.

Viruses in the era of microplastics and plastispheres: Analytical methods, advances and future directions.

Research on microplastics and plastispheres now incorporates the study of viruses to evaluate their effects on the environment and human health. Sharing these new discoveries with the scientific community is crucial to fostering further research and collaborations. However, the current research and methodologies used are fragmented. To address this issue, this paper reviews the literature and the use of methodology developments in each study, identifying four emerging research areas: (1) viral interactions with microplastics; (2) viral population, diversity, and function in plastispheres; (3) the effects of viruses and plastic particles in host-associated environments; and (4) the impacts of viruses within plastispheres. To that end, the article is structured to streamline navigation and help readers easily access existing approaches, recent advancements, key findings, challenges, and opportunities in these areas. Our synthesis reveals that research methods include biochemical assays, omics techniques, spectroscopic analysis, and molecular and bioinformatic tools. Various mechanisms enable viruses to attach to microplastics and plastispheres, leading to widespread distribution and contributing to toxic effects and gene transfer. While the growing evidence is intriguing, there is still much to uncover about their ecological interactions, functions, and impacts.

Interplay between Lung Diseases and Viral Infections: A Comprehensive Review.

The intricate relationship between chronic lung diseases and viral infections is a significant concern in respiratory medicine. We explore how pre-existing lung conditions, including chronic obstructive pulmonary disease, asthma, and interstitial lung diseases, influence susceptibility, severity, and outcomes of viral infections. We also examine how viral infections exacerbate and accelerate the progression of lung disease by disrupting immune responses and triggering inflammatory pathways. By summarizing current evidence, this review highlights the bidirectional nature of these interactions, where underlying lung diseasesincrease vulnerability to viral infections, while these infections, in turn, worsen the clinical course. This review underscores the importance of preventive measures, such as vaccination, early detection, and targeted therapies, to mitigate adverse outcomes in patients with chronic lung conditions. The insights provided aim to inform clinical strategies that can improve patient management and reduce the burden of chronic lung diseases exacerbated by viral infections.

The Prevalence and Associated Factors of Hepatitis B and C Virus in Hemodialysis Patients in Ibb Governorate, Yemen.

Introduction Hepatitis C virus (HCV) and hepatitis B virus (HBV) among patients receiving hemodialysis (HD) remain a major public health problem. However, information is limited about these infections among HD patients in Yemen. This study aimed to estimate the prevalence and associated risk factors of HBV and HCV infections among HD patients in the Ibb governorate, Yemen, and to identify the risk factors for infections in such patients. Methods A cross-sectional study of 374 patients with renal failure who regularly underwent HD at the Al-Thawra Hospital in Ibb city, Yemen, was performed after they agreed to participate and signed informed consent. Enzyme-linked immunoassay (ELISA) was used to test the serum levels of anti-HCV antibodies and hepatitis B surface antigen (HBsAg). Patient data (demographic characteristics and risk factors) were collected via an interview questionnaire and medical records. Logistic regression analysis and chi-square tests were used to analyze the results. Results The overall prevalence of HCV was 31% (n=116), whereas that of HBV was 6.15% (n=23). Three (0.8%) patients had both HCV and HBV. The logistic regression analysis revealed a significant association between an increased number of units of blood transfused (OR = 3.1; 95% CI: 1.7-5.6; p < 0.001), a long duration of dialysis (OR = 3.2; 95% CI: 1.9-5.2; p = 0.001), and HCV infection in HD patients. On the other hand, a history of cupping therapy (Hijama) was significantly associated (OR = 3.4; 95% CI: 1.3-8.77; p < 0.011) with HBV infection in HD patients. Conclusion HCV and HBV infections are more common among HD patients in Yemen than in most Middle Eastern countries. However, the current prevalence rates are declining compared with previously published data among Yemeni HD patients. The duration of HD and number of blood units were independent risk factors for HCV infection, and patients with a history of cupping were identified as having an increased risk of HBV infection. These findings underscore the need to implement strict infection control in HD units.

Characterization of Nasal Mucosal T Cells in Horses and Their Response to Equine Herpesvirus Type 1.

Equine herpesvirus type 1 (EHV-1) enters through the upper respiratory tract (URT). Mucosal immunity at the URT is crucial in limiting viral infection and morbidity. Here, intranasal immune cells were collected from horses (n = 15) during an experimental EHV-1 infection. CD4+ and CD8+ T cells were the major intranasal cell populations before infection and increased significantly by day six and fourteen post-infection, respectively. Nasal mucosal T cells were further characterized in healthy horses. Compared to peripheral blood mononuclear cells (PBMC), mucosal CD8+ T-cell percentages were elevated, while CD4+ T-cell percentages were similar. A small population of CD4+CD8+ T cells was also recovered from mucosal samples. Within the URT tissue, CD4+ cells predominantly accumulated in the epithelial layer, while most CD8+ cells resided deeper in the mucosa or the submucosa below the basement membrane. In vitro stimulation of mucosal cells from healthy horses with (n = 5) or without (n = 5) peripheral T-cell immunity against EHV-1 induced IFN-γ production in nasal T cells upon polyclonal stimulation. However, after EHV-1 re-stimulation, mucosal T cells failed to respond with IFN-γ. This work provided the first characterization of mucosal T-cell phenotypes and functions in the URT of healthy horses and during EHV-1 infection.

Association between Respiratory Virus Infection and Development of De Novo Donor-Specific Antibody in Lung Transplant Recipients.

Chronic lung allograft dysfunction (CLAD) is the most common cause of long-term lung allograft failure. Several factors, including respiratory virus infection (RVI), have been associated with CLAD development, but the underlying mechanisms of these associations are not well understood. We hypothesize that RVI in lung transplant recipients elicits the development of donor-specific antibodies (DSAs), thus providing a mechanistic link between RVI and CLAD development. To test this hypothesis, we retrospectively evaluated for the presence of HLA antibodies in a cohort of lung transplant recipients with symptomatic RVI within the first four months post-transplant using sera at two time points (at/directly after the transplant and following RVI) and time-matched controls without RVI (post-transplant). We found a trend toward the development of de novo DSAs in those with symptomatic RVI versus controls [6/21 (29%) vs. 1/21 (5%), respectively, p = 0.09]. No cases or controls had DSA at baseline. We also found increased rates of CLAD and death among those who developed class II DSA versus those who did not (CLAD: 5/7 (71.4%) vs. 19/34 (54.3%), death: 5/7 (71.4%) vs. 17/35 (48.6%)). Prospective studies evaluating the temporal development of DSA after RVI in lung transplant patients and the subsequent outcomes are warranted.

Immune-Mediated Necrotizing Myopathy With Anti-3-Hydroxy-3-Methylglutaryl-CoA Reductase (HMGCR) Antibodies Following Viral Infection and Without Association With Statin Use: A Case Report.

Immune-mediated necrotizing myopathy (IMNM) is a rare form of inflammatory myopathy characterized by severe muscle weakness, elevated serum creatine kinase (CK) levels, and myofiber necrosis with minimal inflammatory infiltrates. IMNM is frequently associated with autoantibodies, particularly anti-3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), and is often linked to statin use. However, it can also develop in statin-naïve patients, especially following viral infections. We present the case of a 47-year-old woman who developed anti-HMGCR-positive IMNM without prior statin exposure, following a viral respiratory infection and subsequent dengue fever. She initially presented with proximal muscle weakness and elevated CK levels, which worsened after contracting dengue. Diagnostic testing confirmed the presence of anti-HMGCR antibodies, and a muscle biopsy revealed necrotizing myopathy. Treatment with methylprednisolone, intravenous immunoglobulin, and rituximab resulted in significant clinical improvement. This case underscores the need to consider IMNM in patients with unexplained muscle weakness and elevated CK levels, even in the absence of statin use. Viral infections may trigger IMNM, highlighting the importance of early recognition and aggressive immunosuppressive therapy to prevent severe complications. Further research is required to better understand the pathophysiology of IMNM and optimize treatment approaches.

Polarization of the memory B cell response.

Memory B cells are long-lived cells that are induced following infection or vaccination. Upon antigen re-encounter, memory B cells rapidly differentiate into antibody-secreting or germinal center B cells. While memory B cells are an important component of long-term protective immunity following vaccination, they also contribute to the progression of diseases such as autoimmunity and allergy. Numerous subsets of memory B cells have been identified in mice and humans that possess important phenotypic and functional differences. Here, we review the transcriptional circuitry governing memory B cell differentiation and function. We then summarize emerging evidence that the inflammatory environment in which memory B cells develop has an important role in shaping their phenotype and examine the pathways regulating the development of memory B cells during a type 1-skewed and type-2 skewed immune response.

Host combats porcine reproductive and respiratory syndrome virus infection at non-coding RNAs level.

Porcine reproductive and respiratory syndrome virus (PRRSV) poses a significant threat to the global swine industry. The emergence of new, highly virulent strains has precipitated recurrent outbreaks worldwide, underscoring the ongoing battle between host and virus. Thus, there is an imperative to formulate a more comprehensive and effective disease control strategy. Studies have shown that host non-coding RNA (ncRNA) is an important regulator of host - virus interactions in PRRSV infection. Hence, a thorough comprehension of the roles played by ncRNAs in PRRSV infection can augment our understanding of the pathogenic mechanisms underlying PRRSV infection. This review focuses on elucidating contemporary insights into the roles of host microRNAs (miRNAs), long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs) in PRRSV infection, providing both theoretical foundations and fresh perspectives for ongoing research into the mechanisms driving PRRSV infection and its pathogenesis.

Equine coronavirus infection and replication in equine intestinal enteroids.

In this study, equine intestinal enteroids (EIEs) were generated from the duodenum, jejunum, and ileum and inoculated with equine coronavirus (ECoV) to investigate their suitability as in vitro models with which to study ECoV infection. Immunohistochemistry revealed that the EIEs were composed of various cell types expressed in vivo in the intestinal epithelium. Quantitative reverse-transcription PCR (qRT-PCR) and virus titration showed that ECoV had infected and replicated in the EIEs. These results were corroborated by electron microscopy. This study suggests that EIEs can be novel in vitro tools for studying the interaction between equine intestinal epithelium and ECoV.