ADV Infection Research Articles

Adenovirus expressing nc886, an anti-interferon and anti-apoptotic non-coding RNA, is an improved gene delivery vector

Recombinant adenovirus (rAdV) vector is the most promising vehicle to deliver an exogenous gene into target cells and is preferred for gene therapy. Exogenous gene expression from rAdV is often too inefficient to induce phenotypic changes and the amount of administered rAdV must be very high to achieve a therapeutic dose. However, it is often hampered because a high dose of rAdV is likely to induce cytotoxicity by activating immune responses. nc886, a 102-nucleotide non-coding RNA that is transcribed by RNA polymerase III, acts as an immune suppressor and a facilitator of AdV entry into the nucleus. Therefore, in this study, we have constructed an rAdV expressing nc886 (AdV:nc886), to explore whether AdV:nc886 overcomes the aforementioned drawbacks of conventional rAdV vectors. When infected into mouse cell lines and mice, AdV:nc886 expresses a sufficient amount of nc886, which suppresses the induction of interferon-stimulated genes and apoptotic pathways triggered by AdV infection. As a result, AdV:nc886 is less cytotoxic and produces more rAdV-delivered gene products, compared with the parental rAdV vector lacking nc886. In conclusion, this study demonstrates that the nc886-expressing rAdV could become a superior gene delivery vehicle with greater safety and higher efficiency for in vivo gene therapy.

Liver injury and prolonged hospitalization as indicators of severity in patients with adenovirus infections

BackgroundAdenovirus (ADV) is a prevalent infective virus in children, accounting for around 5–10% of all cases of acute respiratory illnesses and 4–15% of pneumonia cases in children younger than five years old. Without treatment, severe ADV pneumonia could result in fatality rates of over 50% in cases of emerging strains or disseminated disease. This study aims to uncover the relationship of clinical indicators with primary ADV infection severity, regarding duration of hospitalization and liver injury.MethodsIn this retrospective study, we collected and analyzed the medical records of 1151 in-patients who met the inclusion and exclusion criteria. According to duration of hospitalization, all patients were divided into three groups. Then the difference and correlation of clinical indicators with ADV infection were analyzed, and the relationship among liver injury, immune cells and cytokines was evaluated.ResultsThe study revealed that patients with a duration of hospitalization exceeding 14 days had the highest percentage of abnormalities across most indicators. This was in contrast to the patients with a hospitalization duration of either less than or equal to 7 days or between 7 and 14 days. Furthermore, correlation analysis indicated that a longer duration of body temperature of ≥ 39°C, bilateral lung lobes infiltration detected by X ray, abnormal levels of AST, PaO2, and SPO2, and a lower age were all predictive of longer hospital stays. Furthermore, an elevated AST level and reduced liver synthesis capacity were related with a longer hospital stay and higher ADV copy number. Additionally, AST/ALT was correlated positively with IFN-γ level and IFN-γ level was only correlated positively with CD4+ T cells.ConclusionsThe study provided a set of predicting indicators for longer duration of hospitalization, which responded for primary severe ADV infection, and elucidated the possible reason for prolonged duration of hospitalization attributing to liver injury via higher ADV copy number, IFN-γ and CD4+ T cells, which suggested the importance of IFN-γ level and liver function monitoring for the patients with primary severe ADV infection.

Adenovirus and Mycoplasma pneumoniae co-infection as a risk factor for severe community-acquired pneumonia in children.

To investigate the pathogenic characteristics and risk factors of pediatric severe community-acquired pneumonia (CAP). We retrospectively analyzed the clinical data of hospitalized children with severe CAP from April 2014 to June 2019 in China. Data of age, sex and pathogenic results were collected: bacterial and fungal cultures, respiratory viruses from sputum or bronchoalveolar lavage fluid (BALF), serum Mycoplasma pneumoniae (MP)-IgM and Chlamydia Pneumoniae-IgM, and BALF or blood (1-3)-β-D-glucan/galactomannan test. A total of 679 children with severe CAP were included in the analysis. The number of cases infected with MP was higher in males than in females. There were significant differences between the ≤1-year and >1-year groups in terms of pathogen. The top three bacteria cultured were Haemophilus influenzae (57/679, 8.4%), Streptococcus pneumoniae (50/679, 7.4%), and Pseudomonas aeruginosa (25/679, 3.7%). The top three viruses detected were adenovirus (AdV, 124/679, 18.3%), respiratory syncytial virus (24/679, 3.5%), and parainfluenza virus (21/679, 3.1%). AdV and MP were the leading pathogens, detected in 18.3% and 32.6% cases, respectively. MP infection increased the risk of AdV infection (OR 3.77, p < 0.0001). MP infection was a risk factor for severe AdV-infected pneumonia, while sex, age, bacteria, Chlamydia Pneumoniae, fungal, and AdV infections were risk factors for severe MP-infected pneumonia. AdV and MP were dominant pathogens in children with severe CAP. AdV and MP infection predisposed each other to develop severe illness. AdV-MP co-infection may lead to severe pneumonia.

In Vitro Antiviral and Anti-Inflammatory Activities of N-Acetylglucosamine: Development of an Alternative and Safe Approach to Fight Viral Respiratory Infections.

Viral respiratory tract infections (RTIs) are responsible for significant morbidity and mortality worldwide. A prominent feature of severe respiratory infections, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, is the cytokine release syndrome. Therefore, there is an urgent need to develop different approaches both against viral replication and against the consequent inflammation. N-acetylglucosamine (GlcNAc), a glucosamine (GlcN) derivative, has been developed as an immunomodulatory and anti-inflammatory inexpensive and non-toxic drug for non-communicable disease treatment and/or prevention. Recent studies have suggested that GlcN, due to its anti-inflammatory activity, could be potentially useful for the control of respiratory virus infections. Our present study aimed to evaluate in two different immortalized cell lines whether GlcNAc could inhibit or reduce both viral infectivity and the inflammatory response to viral infection. Two different viruses, frequent cause of upper and lower respiratory tract infections, were used: the H1N1 Influenza A virus (IAV) (as model of enveloped RNA virus) and the Human adenovirus type 2 (Adv) (as model of naked DNA virus). Two forms of GlcNAc have been considered, bulk GlcNAc and GlcNAc in nanoform to overcome the possible pharmacokinetic limitations of GlcNAc. Our study suggests that GlcNAc restricts IAV replication but not Adv infection, whereas nano-GlcNAc inhibits both viruses. Moreover, GlcNAc and mainly its nanoformulation were able to reduce the pro-inflammatory cytokine secretion stimulated by viral infection. The correlation between inflammatory and infection inhibition is discussed.

2161. Posoleucel, an Allogeneic, Off-the-Shelf Multi-Virus Specific T Cell Therapy, for Severe, Drug-Refractory Viral Infections in Pediatric Patients Following HCT: Results from a Phase 2 Trial

Abstract Background Serious viral infections are a leading cause of morbidity and mortality in pediatric recipients of allogeneic hematopoietic cell transplant (allo-HCT). Most available antivirals have significant toxicities and suboptimal efficacy. Posoleucel (PSL) is an allogeneic, off-the-shelf multi-virus specific T cell investigational product to prevent or treat 6 viral infections (AdV, BKV, CMV, EBV, HHV-6, and JCV). In the phase 2 CHARMS trial of PSL in allo-HCT recipients with intractable viral infections, 95% of patients had a clinical response. Here we present the efficacy and safety data in pediatric patients (&amp;lt; 18 years) enrolled in CHARMS. Methods Allo-HCT patients who had failed antiviral therapy or were unable to tolerate antivirals to control BKV, CMV, AdV, EBV, HHV-6, and/or JCV infection received a single PSL infusion of 2×107 cells/m2. Patients with a response could receive up to 4 more doses after 4 weeks, at 2-week intervals. The primary endpoint of the study was safety and feasibility. Other endpoints included complete and partial clinical response, with complete response (CR) as return to normal range and resolution of signs/symptoms and partial response (PR) defined as ≥50% decrease in viral load and/or ≥50% improvement of clinical signs/symptoms assessed by 6 weeks post infusion. Results 18 pediatric patients received PSL (1 was treated for 2 infections separately). Of the 13 with 1 viral infection, 6 (46%) had BKV, 3 (23%) CMV, 2 (15%) AdV, 2 (15%) HHV-6, and none EBV or JCV. Five (38%) had &amp;gt;1 infection: 3 had 2 infections, 1 had 3 infections, and 1 had 1 viral infection at each of 2 enrollments. 56% of patients received one infusion. PSL was generally well tolerated. Besides fever in 7 patients, we observed no toxicities or symptoms of CRS. Five cases of acute graft-versus-host disease (aGVHD) were observed; 3 in patients with history of aGVHD (2 had aGVHD at baseline). Both de novo cases of GVHD were Grade 1. 100% of patients achieved a clinical response (CR or PR) by 6 weeks including all 5 patients infected with multiple target viruses. Four of 6 patients with AdV infection had CR. Conclusion PSL treatment was well tolerated. All pediatric patients receiving PSL had a clinical response. Phase 3 studies including pediatric patients are in progress. Disclosures Ifigeneia Tzannou, MD, AlloVir: Advisor/Consultant Carlos Ramos, MD, Novartis: Advisor/Consultant Swati Naik, MD, Bellicum: Travel, Accomodation, Expenses Iain Fraser, MD, DPhil, AlloVir: Employee|AlloVir: Stocks/Bonds Marshelle Warren, MD, AlloVir: Advisor/Consultant Badrish Patel, MD, AlloVir: Employee|AlloVir: Stocks/Bonds Dany Ward, RN, AlloVir: Employee|AlloVir: Stocks/Bonds Ann Leen, PhD, AlloVir: Advisor/Consultant|AlloVir: Stocks/Bonds Bilal Omer, MD, AlloVir: Grant/Research Support.

92. Incidence of Adenovirus Respiratory Infection and Coinfection in a Longitudinal Birth Cohort

Abstract Background Adenoviruses (AdVs) are a common cause of acute respiratory illness (ARI) in children and are often detected with other viruses (coinfection). However, the community incidence of AdV infections is poorly understood due to a lack of prospective studies outside the hospital setting. Here, we aim to characterize respiratory AdV infections and coinfections in a prospective birth cohort of children. Methods The PREVAIL cohort is a CDC-funded, 2-year birth cohort, conducted from 2017–2020 in Cincinnati, OH. ARI was defined as the presence of cough or fever identified with weekly maternal text surveys and medical chart review. Mid-turbinate nasal swabs were collected weekly. Swabs were tested using Luminex Respiratory Pathogen Panel. AdV infection was defined as a swab positive for AdV and included subsequent positives &amp;lt; 30 days apart. Coinfection was defined as detection of any other virus(es) during an AdV infection. Children who submitted at least 70% of weekly samples were included in our analysis. Results 101 children met inclusion criteria, representing 165 child-years. 137 distinct AdV infections were identified (incidence 0.84 infections per child year), with 98 (97%) children having ≥1 AdV infection. Only 40% (n=55) of AdV infections were symptomatic. Of those with symptomatic infections, 51% (n=28) sought medical care, with 42% (n=23) presenting to a primary care provider and 9% (n=5) resulting in an ED visit or hospital admission. Coinfections were detected in 67% (n=92) of AdV infections, with 45% (n=62) coinfected with 1 virus, 19% (n=26) with 2 viruses, and 3% (n=4) with ≥3 viruses. 77% of coinfections (n=71) were rhino/enterovirus. The number of coinfections or the specific coinfection virus was not associated with an increase in symptom prevalence or symptom severity (all p &amp;gt; 0.05). Viral Coinfection Frequency with Adenovirus Infection in the PREVAIL Cohort. Adenovirus Infection and Coinfection in the PREVAIL Cohort. Conclusion In this cohort of healthy children, AdVs were a common cause of respiratory infection. Most infections were asymptomatic or resulted in mild symptoms. Two-thirds of AdV infections involved viral coinfections, but coinfection was not associated with more frequent or severe symptoms. Our findings suggest studies that only include symptomatic or hospitalized patients may overestimate AdVs disease severity. Disclosures All Authors: No reported disclosures.

Trained innate immunity and human adenovirus infection as a novel mechanism linked to the severe acute hepatitis of unknown origin in children during the COVID‐19 pandemic

Key points Recent clinical findings indicate that in severe acute hepatitis of unknown origin in children, the innate immune response underlies the main pathogenesis, regardless of the involved pathogens, including adenoviruses (AdVs) that predominantly affect the gut. The present work suggests that in the presence of trained immunity, the inflammation triggered by AdV infection is mild and confined to the gut, whereas without trained immunity, AdV infection can lead to viremia, severe inflammation, and even cytokine storms.

Newly recruited intraepithelial Ly6A+CCR9+CD4+ T cells protect against enteric viral infection

The intestinal epithelium comprises the body's largest surface exposed to viruses. Additionally, the gut epithelium hosts a large population of intraepithelial T lymphocytes, or IELs, although their role in resistance against viral infections remains elusive. By fate-mapping Tcells recruited to the murine intestine, we observed an accumulation of newly recruited CD4+ Tcells after infection with murine norovirus CR6 and adenovirus type-2 (AdV), but not reovirus. CR6- and AdV-recruited intraepithelial CD4+ Tcells co-expressed Ly6A and chemokine receptor CCR9, exhibited T helper 1 and cytotoxic profiles, and conferred protection against AdV invivo and in an organoid model in an IFN-γ-dependent manner. Ablation of the Tcell receptor (TCR) or the transcription factor ThPOK in CD4+ Tcells prior to AdV infection prevented viral control, while TCR ablation during infection did not impact viral clearance. These results uncover a protective role for intraepithelial Ly6A+CCR9+CD4+ Tcells against enteric adenovirus.

Adenovirus: Epidemiology, Global Spread of Novel Types, and Approach to Treatment.

Adenoviruses (AdVs) are DNA viruses that typically cause mild infections involving the upper or lower respiratory tract, gastrointestinal tract, or conjunctiva. Rare manifestations of AdV infections include hemorrhagic cystitis, hepatitis, hemorrhagic colitis, pancreatitis, nephritis, or meningoencephalitis. AdV infections are more common in young children, due to lack of humoral immunity. Epidemics of AdV infection may occur in healthy children or adults in closed or crowded settings (particularly military recruits). The vast majority of cases are self-limited. However, the clinical spectrum is broad and fatalities may occur. Dissemination is more likely in patients with impaired immunity (e.g., organ transplant recipients, human immunodeficiency virus infection). Fatality rates for untreated severe AdV pneumonia or disseminated disease may exceed 50%. More than 100 genotypes and 52 serotypes of AdV have been identified and classified into seven species designated HAdV-A through -G. Different types display different tissue tropisms that correlate with clinical manifestations of infection. The predominant types circulating at a given time differ among countries or regions, and change over time. Transmission of novel strains between countries or across continents and replacement of dominant viruses by new strains may occur. Treatment of AdV infections is controversial, as prospective, randomized therapeutic trials have not been done. Cidofovir has been the drug of choice for severe AdV infections, but not all patients require treatment. Live oral vaccines are highly efficacious in reducing the risk of respiratory AdV infection and are in routine use in the military in the United States but currently are not available to civilians.

Evidence of MHC class I and II influencing viral and helminth infection via the microbiome in a non-human primate.

Until recently, the study of major histocompability complex (MHC) mediated immunity has focused on the direct link between MHC diversity and susceptibility to parasite infection. However, MHC genes can also influence host health indirectly through the sculpting of the bacterial community that in turn shape immune responses. We investigated the links between MHC class I and II gene diversity gut microbiome diversity and micro- (adenovirus, AdV) and macro- (helminth) parasite infection probabilities in a wild population of non-human primates, mouse lemurs of Madagascar. This setup encompasses a plethora of underlying interactions between parasites, microbes and adaptive immunity in natural populations. Both MHC classes explained shifts in microbiome composition and the effect was driven by a few select microbial taxa. Among them were three taxa (Odoribacter, Campylobacter and Prevotellaceae-UCG-001) which were in turn linked to AdV and helminth infection status, correlative evidence of the indirect effect of the MHC via the microbiome. Our study provides support for the coupled role of MHC diversity and microbial flora as contributing factors of parasite infection.

Safety and Efficacy of Tcralpha/Beta and CD19 Depleted Haploidentical Stem Cell Transplantation Following Reduced Intensity Conditioning in Children: Results of a Prospective Multicenter Phase I/II Clinical Trial

We report one year follow-up data on safety and efficacy from the first prospective, multi-center, open-label, single-arm phase I/II clinical trial of TCRalpha/beta and CD19-depleted haploidentical stem cells grafts which were generated with the CliniMACS plus System (Miltenyi Biotec, Germany) and transplanted after reduced-intensity conditioning in pediatric patients with various malignant and non-malignant diseases (www.clinicaltrialsregister.org; 2011-005562-38). 30 patients were enrolled in the pediatric cohort of the trial and received single agent MMF as short-term GVHD prophylaxis (40mg/kg/day for 30 days). The grafts were manufactured in 4 GMP sites. The kinetics of immune cell reconstitution were measured in two core labs using standardized phenotypic analyses and the MACSQuant flow cytometer (Miltenyi Biotec, Germany). We previously reported the outcome of the first 100 days. This report covers a median follow-up of 367 days.Results: Thirty patients from six hospitals were treated (13 female, 17 male; median age 7 years, range 1 - 17 years). 10 patients had ALL, 8 AML, 6 solid tumors (soft tissue sarcomas and neuroblastomas), 3 MDS/MPS, and 1 each with lysosomal storage disorder, SCID, and Wiskott Aldrich syndrome. Disease status in acute leukemias/MDS were: CR1 (n=4), relapsed/refractory (n=17). 5/6 patients with solid tumors had relapsed metastatic disease. The conditioning regimen consisted of 15 or 30 mg ATG (Fresenius/Grafalon) or 7 Gy total nodal irradiation, 160 mg/m2 fludarabine, 10 mg/kg thiotepa, and 140 mg/m2 melphalan. The median numbers of CD34+ cells, TCRab+ cells and CD20+ cells infused per kg bw were 14.6 x 106 (range, 4 - 54.9), 14 x 103 (range, 0.62 - 40.6) and 0.55 x 105 (range, 0.04 - 1.85), respectively. Main cellular components of the graft were NK and TCRgd+ cells (median 6.67 x 107 and 1.58 x 107 cells per kg bw, respectively) with consistently high log depletions of TCRab+ and CD19+ cells: 4.75 and 3.44, respectively.None of the 30 recipients developed severe acute GVHD grades III - IV. The vast majority of patients (97%) experienced no or only grade I acute GVHD despite minimal GVHD prophylaxis after transplantation. One patient had acute GVHD grade II. Chronic GVHD was observed in two out of 23 patients (9%): one with skin and one with gut/liver involvement. In both cases GVHD was extensive.Twenty-four patients were evaluable for assessment of immune reconstitution until 1 year or discontinuation. Figure 1 shows fast and stable reconstitution of T, B and NK cells. TCRab+ T cells continued to increase during the observation period up to a median value of 741 cells/µL (range, 245 - 3132), whereas TCRgd+ T cells remained stable after reaching a plateau around day 20.ADV reactivation in 16 patients contributed most to infectious complications following transplantation. Of these 6 patients developed ADV disease. Within the first year after transplantation 3 patients died of overwhelming of ADV infection. One EBV reactivation was observed but recovered completely. 8 patients experienced CMV reactivation after transplantation. 7 of 8 patients recovered. Another 2 patients had CMV reactivation prior to transplant, but recovered shortly after engraftment. No CMV disease was observed within the first year. 6 patients had bacterial sepsis but recovered. In one patient CMV reactivation continued until death due to sepsis after graft failure.At a median of 367 days (range, 14 - 809) among the 30 recipients, 16 are alive at time of last follow-up, 8 died of relapse, 3 died of ADV infection, 1 died of sepsis after graft failure and 1 died of ARDS. 1 patient is lost to follow-up. Of the 8 patients who died of relapsed disease, 4 had active disease at time of transplant, 2 were in remission and 2 had disease status not evaluable. Cumulative incidences of relapse and NRM were 27% and 16% at one year, respectively. Kaplan-Meyer estimates of overall survival and DFS were 64% and 60% at one year, respectively.Conclusion: The CliniMACS TCRab+ / CD19+ depletion system yielded a large number of CD34+ cells, NK cells and TCRgd+ cells, that could be infused safely into pediatric patients with minimal risk of severe acute and chronic GVHD. The immune reconstitution was rapid. Coupled with a reduced-intensity regimen, the overall TRM was low. However, further efforts have to be made to avoid ADV associated infections. [Display omitted] DisclosuresLang:Miltenyi Biotec GmbH: Patents & Royalties, Research Funding. Bader:Novartis, Medac, Amgen, Riemser, Neovii: Consultancy, Honoraria, Research Funding. Karitzky:Miltenyi Biotec GmbH: Employment. Holtkamp:Miltenyi Biotec GmbH: Employment. Siewert:Miltenyi Biotec GmbH: Employment. Pflitsch:Miltenyi Biotec GmbH: Employment. Handgretinger:Miltenyibiotec GmbH: Patents & Royalties: Co-patent holder of TcR alpha/beta depletion technologies, Research Funding.

ML-AdVInfect: A Machine-Learning Based Adenoviral Infection Predictor

Adenoviruses (AdVs) constitute a diverse family with many pathogenic types that infect a broad range of hosts. Understanding the pathogenesis of adenoviral infections is not only clinically relevant but also important to elucidate the potential use of AdVs as vectors in therapeutic applications. For an adenoviral infection to occur, attachment of the viral ligand to a cellular receptor on the host organism is a prerequisite and, in this sense, it is a criterion to decide whether an adenoviral infection can potentially happen. The interaction between any virus and its corresponding host organism is a specific kind of protein-protein interaction (PPI) and several experimental techniques, including high-throughput methods are being used in exploring such interactions. As a result, there has been accumulating data on virus-host interactions including a significant portion reported at publicly available bioinformatics resources. There is not, however, a computational model to integrate and interpret the existing data to draw out concise decisions, such as whether an infection happens or not. In this study, accepting the cellular entry of AdV as a decisive parameter for infectivity, we have developed a machine learning, more precisely support vector machine (SVM), based methodology to predict whether adenoviral infection can take place in a given host. For this purpose, we used the sequence data of the known receptors of AdVs, we identified sets of adenoviral ligands and their respective host species, and eventually, we have constructed a comprehensive adenovirus–host interaction dataset. Then, we committed interaction predictions through publicly available virus-host PPI tools and constructed an AdV infection predictor model using SVM with RBF kernel, with the overall sensitivity, specificity, and AUC of 0.88 ± 0.011, 0.83 ± 0.064, and 0.86 ± 0.030, respectively. ML-AdVInfect is the first of its kind as an effective predictor to screen the infection capacity along with anticipating any cross-species shifts. We anticipate our approach led to ML-AdVInfect can be adapted in making predictions for other viral infections.

E3 ubiquitin ligase Mindbomb 1 facilitates nuclear delivery of adenovirus genomes.

The journey from plasma membrane to nuclear pore is a critical step in the lifecycle of DNA viruses, many of which must successfully deposit their genomes into the nucleus for replication. Viral capsids navigate this vast distance through the coordinated hijacking of a number of cellular host factors, many of which remain unknown. We performed a gene-trap screen in haploid cells to identify host factors for adenovirus (AdV), a DNA virus that can cause severe respiratory illness in immune-compromised individuals. This work identified Mindbomb 1 (MIB1), an E3 ubiquitin ligase involved in neurodevelopment, as critical for AdV infectivity. In the absence of MIB1, we observed that viral capsids successfully traffic to the proximity of the nucleus but ultimately fail to deposit their genomes within. The capacity of MIB1 to promote AdV infection was dependent on its ubiquitination activity, suggesting that MIB1 may mediate proteasomal degradation of one or more negative regulators of AdV infection. Employing complementary proteomic approaches to characterize proteins proximal to MIB1 upon AdV infection and differentially ubiquitinated in the presence or absence of MIB1, we observed an intersection between MIB1 and ribonucleoproteins (RNPs) largely unexplored in mammalian cells. This work uncovers yet another way that viruses utilize host cell machinery for their own replication, highlighting a potential target for therapeutic interventions that counter AdV infection.

Adenovirus Infection-associated Central Nervous System Disease in Children.

Adenovirus (Adv) is a frequent etiology of acute respiratory tract infections. Although rare, neurologic manifestations are known to occur during Adv infection. We retrospectively analyzed clinical, laboratory, outcome and the relationship between clinical characteristics and viral detection results in the cerebrospinal fluid (CSF) in children with Adv-associated central nervous system (CNS) dysfunction. (1.5%) cases had Adv-associated CNS manifestations. The median age was 1.4 years and 20 (95%) were less than 5 years of age. Six (28%) were male. The most frequently cited CNS symptoms were altered consciousness (100%) and seizure (14.3%). Fourteen cases (73.7%) had abnormal electroencephalogram examination and 6 cases (37.5%) had abnormal imaging. None of the patients had received cidofovir administration. Twenty children recovered without sequelae and 1 patient died of respiratory failure. Patients with positive Adv polymerase chain reaction (n = 11) presented lower onset age compared with that of patients with negative Adv polymerase chain reaction (n = 10) in the CSF. Clinical manifestation, laboratory findings, imaging studies and electroencephalogram showed no significant difference between the 2 groups. Adv is a rare cause of CNS disease in children, mainly causing altered consciousness. Adv was detected in more cases in the respiratory tract than the CSF, but the majority of patients had the virus detected in both. The lack of Adv in the CSF does not exclude CNS involvement. Furthermore, the viral detection results in the CSF do not seem useful as an indicator of the severity of CNS disease.

Treatment of Severe, Drug-Refractory Viral Infections with Allogeneic, Off-the-Shelf Multi-Virus Specific T Cells in Patients Following HSCT: Results from a Phase 2 Study

Background. Serious viral infections associated with delayed immune reconstitution are a leading cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). In approximately 90% of allogeneic HSCT patients, the suppressed immune system allows viruses that were previously in a latent, quiescent state to reactivate, with up to 60% of patients experiencing a reactivation of 2 or more viruses, including BKV, CMV, AdV, EBV and HHV-6; HSCT also predisposes for persistent acute infections, such as AdV. There is a lack of effective antiviral drugs to treat the majority of these infections and, when used clinically, available antiviral therapies are associated with significant toxicities, resulting in a substantial unmet medical need in this population. Viralym-M is an allogeneic, off-the-shelf multi-virus specific T cell (VSTs) product targeting five common viruses (BKV, CMV, AdV, EBV and HHV-6) that can lead to devastating diseases in allogeneic HSCT recipients. As T cells play a central role in response to viral infection, Viralym-M is intended to restore VST immunity to treat or prevent viral diseases and infections. Efficacy and safety of Viralym-M in allogeneic HSCT recipients with at least one treatment-refractory infection were evaluated in a phase II proof-of-concept trial (CHARMS study). In a previously published interim analysis from the study, infusion of Viralym-M was well tolerated and associated with clinical responses in most patients. Here we present the efficacy and safety data from the complete cohort of patients enrolled in the CHARMS study. Methods. Allogeneic HSCT patients who had either failed antiviral therapy or were unable to tolerate standard antivirals to control BKV, CMV, AdV, EBC, HHV-6, and/or JCV infection were enrolled to the study. Viralym-M was given as a single infusion of 2x107 VSTs/m2. If a partial response was achieved, patients could receive up to 4 additional doses after 4 weeks, at 2-week intervals. Study end points included resolution of the target infections as measured by viral load, resolution of clinical signs and symptoms as determined by the primary investigator, and safety. A complete response (CR) was defined as viral load returning to normal range and resolution of clinical signs/symptoms, while a partial response (PR) as a ≥50% decrease in viral load and/or 50% improvement of clinical signs/symptoms. Clinical end points were assessed by 6 weeks post infusion, and patients were followed until the end of study at 12 months. Persistence of the VSTs was examined by the epitope specificity of circulating T cells. Results. Fifty-eight patients were treated with Viralym-M, with one patient who was treated for two different infections separately (N=59). Nineteen (32.2%) were pediatric and 40 (67.8%) were adults. Of the patients with one viral infection, 30.5% had BKV, 28.8% CMV, 13.6% AdV, 5.1% HHV-6, 1.7% EBV, and 1.7% JCV. Eleven patients (18.6%) had more than one virus infection: 10 were co-infected with 2 different viruses, and 1 with 3 different viruses. The majority of patients received one infusion (74.6%). Overall, more than 90% of patients achieved a clinical response (PR or CR) by 6 weeks post Viralym-M infusion. 11 patients on the trial were co-infected with multiple viruses and 19 of the 23 viral infections in these patients responded to Viralym-M. We tested for persistence in 16 patients and 11 (69%) had circulating VSTs with persistence of VSTs of third party origin for up to 12 weeks. Treatment with Viralym-M was generally well tolerated. No immediate toxicities were observed and, other than isolated fever in a few patients, we observed no symptoms associated with CRS. Fourteen cases of acute graft-versus-host disease (GVHD) were observed; 8 of these were pre-existing. Of the six patients that experienced de novo GVHD, all had transient grade 1 skin GVHD that resolved with treatment. Conclusions. More than 90% of patients achieved a clinical response by 6 weeks post Viralym-M treatment, which was generally well tolerated, suggesting that Viralym-M can be safely and effectively used to treat viral infections, including multiple infections, in patients following allogeneic HSCT. Viralym-M has now received Regenerative Medicine Advanced Therapy (RMAT) and PRIority MEdicine (PRIME) designation from both the FDA and EMA. Phase 3 and proof-of-concept studies are in progress. Disclosures Tzannou: Allovir: Consultancy. Watanabe:AlloVir: Consultancy. Kuvalekar:AlloVir: Consultancy. Ramos:Novartis: Membership on an entity's Board of Directors or advisory committees; Tessa Therapeutics: Research Funding; Kuur Therapeutics: Research Funding. Heslop:Novartis: Consultancy; Gilead Biosciences: Consultancy; PACT Pharma: Consultancy; Kiadis: Consultancy; Tessa Therapeutics: Consultancy, Research Funding; Marker Therapeutics: Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees; AlloVir: Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees. Leen:AlloVir: Consultancy, Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees; Marker Therapeutics: Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees. Omer:Allovir: Consultancy, Research Funding.

Epidemiological study on respiratory syncytial virus-caused respiratory infection in children and preliminary analysis of changes in immune function

Objective To analyze the epidemiological characteristics of respiratory syncytial virus (RSV)-caused respiratory diseases in children in Hebei Province and the changes in peripheral blood lymphocyte subsets and inflammatory indexes. Methods A total of 9 491 sputum specimens and 9 491 paired peripheral blood specimens were collected from children admitted to Hebei Children′s Hospital for respiratory infection in 2018. RSV-positive sputum specimens were screened by multiple detection reagents for 13 kinds of respiratory pathogens. Flow cytometry was uses to detect T and B lymphocytes and NK cells in peripheral blood samples of randomly screened RSV-positive children. Procalcitonin (PCT) was measured by Roche E411 luminescence analyzer. Hypersensitive C-reactive protein (hs-CRP) was detected by Roche Cobas 8000 C701 biochemical analyzer. White blood cells (WBC) were measured by Sysmex XN-BN3 hematometer. Results The positive rate of RSV in children with respiratory diseases was 13.08% in Hebei Province in 2018. There were significant differences in RSV-positive rates among different age groups (χ2=479.297 6, P<0.000 1). The positive rate of RSV decreased gradually with age (χ2=-20.282 7, P<0.000 1) and was higher in male than in female (χ2=34.552 7, P<0.000 1). The incidence of co-infection of RSV with other respiratory pathogens was 29.49% (366/1 241), mainly caused by human rhinovirus (HRV, 150/1 241) and adenovirus (ADV, 40/1 241). The main epidemic seasons of RSV infection were winter and spring. The epidemic trends of simple RSV infection and co-infection were consistent. There were significant differences in inflammatory indexes, WBC (P<0.01), CD4+ (P=0.015) and CD4+ /CD8+ cells (P=0.016) between simple RSV infection and co-infection groups. Conclusions RSV was a common pathogen causing respiratory diseases in children in Hebei Province. The younger the children were, the more likely they were to be infected with RSV. RSV infection was easily complicated by HRV or ADV infection. The epidemic seasons of RSV infection in Hebei were winter and spring. Both simple infection and co-infection of RSV might result in immune dysfunction. Key words: Respiratory syncytial virus; Lymphocyte subsets; Molecular epidemiology; Immunity

Characteristics of the Tongue Coating&amp;nbsp;Microbiome&amp;nbsp;in Patients Infected With Adenovirus

Background: Adenovirus(AdV) infection, which distributes globally when regional epidemics are reported throughout the year, has no specific treatment yet. As the significance of the oral microbiome has been increasingly recognized in virus infection, this study aims to explore the characteristics of the tongue coating microbiome in AdV infected patients for the first time. Methods: 26 patients with AdV infection and 34 matched healthy controls were enrolled to collect tongue coating samples for 16S rRNA gene sequencing. Information on the tongue coating microbiome was annotated by the Human Oral Microbiome Database(HOMD). Results: The characteristics of AdV infection include decreased abundance and evenness of microbiome among individuals (p<0.001) and significant differences in the relative abundance at different levels (phylum, class, order, family, genus) (p<0.05), compared with the control group. It should be noted that the combination of 3 genera, including Neisseria, Fusobacterium and Haemophilus, can be used as potential biomarkers to accurately distinguish AdV infection from controls (AUC:0.9714). In addition, Neisseria and Haemophilus were also associated with platelet, AST and CRE. Fifteen KEGG pathways were found associated with AdV infection which was closely related to three biomarkers, with transporters the most important functional module. Conclusions: This study described the characteristics of tongue coating microbiome of ADV infection, provided not only new evidence for the biomarkers and pathways associated with AdV infection, but also novel insights into pathogenesis, prevention and treatment of AdV infections as well. Funding Statement: This work was supported by the National Natural Science Foundation of China (nos. 81673806), Emergency Scientific Research Project of Beijing administration of traditional Chinese medicine(nos.YJ2020-03), Emergency Research Project (nos.BWS20J006), Medical Research Program(CX19015) and Bio-safety Construction Plan (nos.A3704041902-02). Declaration of Interests: The authors report no conflict of interest, and no commercial or other association that might pose a conflict of interest (e.g., pharmaceutical stock ownership, consultancy, advisory board membership, relevant patents, or research funding). Ethics Approval Statement: The study complied with the Ethical Principles of the World Medical Association Declaration of Helsinki and experimental protocols were approved by the committee on ethics of the 5th Medical Centre of Chinese PLA General Hospital (formerly the 302 Military Hospital).

Investigation on Atypical Pathogens related with Community Acquired Pneumonia and the Factors Associated with Mycoplasma Pneumoniae Infection in Jiangsu, China

Community acquired pneumonia remains a leading infectious cause of hospitalization with substantial morbidity and mortality in China. At present, the role of atypical pathogens in CAP arouses peoples' concern. Previous studies indicated that MP was the dominant pathogen of CAP, but the associated factors of MP infection were rarely reported. We retrospectively investigated the distribution of atypical pathogens related with CAP and compared their differences among various populations. Furthermore, we accessed the associated factors of MP infection in various population. A total of 3,675 patients were enrolled and divided into three groups. One thousand and eighty-nine subjects (29.6%) were infected with at least one atypical pathogen. MP was the most predominant pathogen in these CAP patients. Our study found that infection rates of the atypical pathogens were significantly different among three groups. Our results also revealed, in a pediatric group, as the temperature increased, so did the infection rate of MP, while it was the opposite in adult and elderly groups. Furthermore, in preschool and school-age children, high temperature, female, PIVs, ADV, and INFB infection were independent risk factors for MP infection, INFA infection was a protection factor for MP infection. However, in adult and elderly groups, the associated factors might be different. The infection of atypical pathogens related with CAP is quite serious, and MP infection plays a key role in CAP. Besides, the infection rates of the atypical pathogens are different in various populations, as are the associated factors for MP infection.

The E3 Ubiquitin Ligase Mind Bomb 1 Controls Adenovirus Genome Release at the Nuclear Pore Complex

Adenoviruses (AdVs) cause respiratory, ocular, and gastrointestinal tract infection and inflammation inimmunocompetent people and life-threatening disease upon immunosuppression. AdV vectors arewidely used in gene therapy and vaccination. Incoming particles attach to nuclear pore complexes (NPCs) of post-mitotic cells, then rupture and deliver viral DNA (vDNA) to the nucleus or misdeliver to the cytosol. Our genome-wide RNAi screen in AdV-infected cells identified the RING-type E3 ubiquitin ligase Mind bomb 1 (Mib1) as a proviral host factor for AdV infection. Mib1 is implicated in Notch-Delta signaling, ciliary biogenesis, and RNA innate immunity. Mib1 depletion arrested incoming AdVs at NPCs. Induced expression of full-length but not ligase-defective Mib1 in knockout cells triggered vDNA uncoating from NPC-tethered virions, nuclear import, misdelivery of vDNA, and vDNA expression. Mib1 is an essential host factor for AdV uncoating in human cells, and it provides a new concept for licensing virion DNA delivery through the NPC.

Cold Weather Viruses.

1. Asif Noor, MD* 2. Theresa Fiorito, MD* 3. Leonard R. Krilov, MD*,† 1. *Department of Pediatrics, Children's Medical Center, NYU Winthrop Hospital, Mineola, NY 2. †Department of Pediatrics, State University of New York, Stony Brook School of Medicine, Stony Brook, NY * Abbreviations: AAP: : American Academy of Pediatrics Adv: : adenovirus CDC: : Centers for Disease Control and Prevention FDA: : Food and Drug Administration hMPV: : human metapneumovirus PCR: : polymerase chain reaction PIV: : parainfluenza virus RSV: : respiratory syncytial virus RV: : rhinovirus Clinicians must learn to identify viral infections in children during the winter months and must practice caution with the use of unnecessary medications in such cases. Recognition of the clinical pattern of viral infection (eg, bronchiolitis) in conjunction with judicious use of viral tests (either office-based immunoassays or newer molecular tests) may assist in epidemiological monitoring, cohorting patients in the hospital, withholding unnecessary therapies, and providing a definitive diagnosis. After completing this article, readers should be able to: 1. Review the epidemiological aspects and clinical signs and symptoms of common cold weather viruses. 2. Recognize situations in which viral testing is indicated. 3. Recognize situations in which treatment is indicated. In early November you are evaluating a 9-month-old boy born at 33 weeks of gestation. The infant presents with 2 days of fevers (101°F–102°F [38.3°C–38.8°C]), copious rhinorrhea, and 1 day of coughing with difficulty breathing. He is otherwise feeding well and has had adequate urination. His 4-year-older sister has an upper respiratory tract infection. On physical examination, the infant has a respiratory rate of 45 breaths/min without chest wall retractions. On auscultation there is good air entry with scattered rhonchi bilaterally. What is the most appropriate next step in management? 1. Obtain respiratory syncytial virus (RSV) and influenza antigen testing. 2. Obtain a chest radiograph to look for focal infiltrate. 3. Provide supportive care with nasal …