Human Adenovirus Research Articles

Initial safety assessment of the endoscopically injected oncolytic virus OBP-301 in medically inoperable esophageal cancer: NRG-GI007.

341 Background: Definitive chemoradiation (CRT) is a standard-of-care for patients (Pts) with medically inoperable esophageal cancer (EC). The NRG/RTOG 0436 study of cisplatin/paclitaxel and RT (+/- cetuximab) as definitive therapy showed that about half of Pts have locally persistent disease. OBP-301 is a conditionally restricted, replication-competent adenovirus derived from human adenovirus type 5 that adds a human Telomerase Reverse Transcriptase (hTERT) gene promoter, replicating only in tumor cells to cause lysis. It may cause immunogenic cell death, enhance RT, and improve local control. OBP-301 was previously studied with RT in Japanese esophageal cancer Pts and shown to be safe and has promising activity.1 Methods: In NRG-GI007, a phase 1 study (NCT04391049), OBP-301 is added to weekly carboplatin/paclitaxel and RT (50.4 Gy/28 fractions) for Pts with medically inoperable EC. Pts receive 1-2mL of intratumoral OBP-301 1×1012virus particles/ml via endoscopy 3 days prior to and then at days 12 and 26 of CRT. The primary endpoint is dose-limiting toxicity (DLT), defined as adverse events (AEs, by CTCAE v5) definitely or probably attributed to OBP-301 that meet either of the following: (1) leading to a >14-day cumulative delay in CRT or (2) any grade ≥ 3 AE EXCEPT: grade 3 nausea/vomiting, grade 3 esophagitis or dehydration, first occurrence of grade 3/4 neutropenia, or grade 3/4 lymphopenia. Initially, 6 evaluable Pts (eligible and started protocol treatment) are enrolled. If protocol defined DLT occurs in ≤1 of 6 Pts, the dose will be deemed safe and an expansion cohort of 9 more will be enrolled to further assess safety and obtain a preliminary assessment of clinical complete response. If ≥2 Pts have a DLT then one de-escalated OBP-301 dose will be assessed. Results: From June 2020 to April 2023, 6 evaluable Pts were enrolled. Median age was 73.5 years, all male. Five Pts had adenocarcinoma and 1 had squamous cell carcinoma; 3 Pts had node-positive disease. All Pts received all planned OBP-301 injections, along with 50.4 Gy RT. Four out of 6 Pts received all 5 weekly doses of chemo for > 85% of planned total dose, and 2 got 4 weekly doses for > 70% of planned total dose. No Pt experienced ³7 day treatment delay. The following treatment-related (attributed as definitely, probably, or possibly related to CRT and/or OBP-301) grade 3 AEs were reported across 5 Pts: decreased neutrophil count (3 Pts), decreased lymphocyte count (2 Pts), decreased WBC (2 Pts), and fever and fatigue (1 Pt each). There were 4 Pts with AEs reported as definitely or probably related to OBP-301, all grade ≤2. No DLT occurred. Conclusions: No DLTs in the 6 evaluable Pts were observed and it is concluded that the initial OBP-301 dose level is safe. NRG-GI007 was reopened on August 7, 2023 to the expansion cohort. Full toxicity and treatment data will be presented. 1. Eur J Cancer 2021;153:98. Clinical trial information: NCT04391049 .

Inhibition of Bromodomain Proteins Enhances Oncolytic HAdVC5 Replication and Efficacy in Pancreatic Ductal Adenocarcinoma (PDAC) Models.

Pancreatic ductal adenocarcinoma (PDAC) is the most aggressive type of pancreatic cancer, which rapidly develops resistance to the current standard of care. Several oncolytic Human AdenoViruses (HAdVs) have been reported to re-sensitize drug-resistant cancer cells and in combination with chemotherapeutics attenuate solid tumour growth. Obstacles preventing greater clinical success are rapid hepatic elimination and limited viral replication and spread within the tumour microenvironment. We hypothesised that higher intratumoural levels of the virus could be achieved by altering cellular epigenetic regulation. Here we report on the screening of an enriched epigenetics small molecule library and validation of six compounds that increased viral gene expression and replication. The greatest effects were observed with three epigenetic inhibitors targeting bromodomain (BRD)-containing proteins. Specifically, BRD4 inhibitors enhanced the efficacy of Ad5 wild type, Ad∆∆, and Ad-3∆-A20T in 3-dimensional co-culture models of PDAC and in vivo xenografts. RNAseq analysis demonstrated that the inhibitors increased viral E1A expression, altered expression of cell cycle regulators and inflammatory factors, and attenuated expression levels of tumour cell oncogenes such as c-Myc and Myb. The data suggest that the tumour-selective Ad∆∆ and Ad-3∆-A20T combined with epigenetic inhibitors is a novel strategy for the treatment of PDAC by eliminating both cancer and associated stromal cells to pave the way for immune cell access even after systemic delivery of the virus.

Neutralizing monoclonal antibodies protect against human adenovirus type 55 infection in transgenic mice and tree shrews

ABSTRACT Re-emerging human adenovirus type 55 (HAdV55) has become a significant threat to public health due to its widespread circulation and the association with severe pneumonia, but an effective anti-HAdV55 agent remains unavailable. Herein, we report the generation of macaque-derived, human-like monoclonal antibodies (mAbs) protecting against HAdV55 infection with high potency. Using fluorophore-labelled HAdV55 virions as probes, we isolated specific memory B cells from rhesus macaques (Macaca mulatta) that were immunized twice with an experimental vaccine based on E1-, E3-deleted, replication-incompetent HAdV55. We cloned a total of 19 neutralizing mAbs, nine of which showed half-maximal inhibitory concentrations below 1.0 ng/ml. These mAbs recognized the hyper-variable-region (HVR) 1, 2, or 7 of viral hexon protein, or the fibre knob. In transgenic mice expressing human desmoglein-2, the major cellular receptor for HAdV55, a single intraperitoneal injection with hexon-targeting mAbs efficiently prevented HAdV55 infection, and mAb 29C12 showed protection at a dose as low as 0.004 mg/kg. Fibre-targeting mAb 28E8, however, showed protection only at a dose up to 12.5 mg/kg. In tree shrews that are permissive for HAdV55 infection and disease, mAb 29C12 effectively prevented HAdV55-caused pneumonia. Further analysis revealed that fibre-targeting mAbs blocked the attachment of HAdV55 to host cells, whereas hexon-targeting mAbs, regardless of their targeting HVRs, mainly functioned at post-attachment stage via inhibiting viral endosomal escape. Our results indicate that hexon-targeting mAbs have great anti-HAdV55 activities and warrant pre-clinical and clinical evaluation.

Human adenovirus-associated health risk in the recreational waters of the Yal-ku lagoon in the Mexican Caribbean.

The study objective was to evaluate human faecal contamination impacts in the Yal-ku lagoon in the Mexican Caribbean and to estimate adenovirus infection and illness risks associated with recreational exposure during water activities. A total of 20 water samples (10 from each site × two sites) (50 L) were collected monthly over a period of 12 months from two selected sampling sites in the swimming area of the Yal-ku lagoon. The occurrence of faecal-associated viruses was explored, and human adenovirus (HAdV) and pepper mild mottle virus (PMMoV) concentrations were quantified. A quantitative microbial risk assessment (QMRA) model was used to estimate exposure and subsequent adenovirus infection and illness risk for 1 h of swimming or snorkelling. Somatic and F + -specific coliphages occurred in 100% of the samples. Both HAdV and PMMoV were detected at a 60% frequency thereby indicating persistent faecal inputs. PMMoV concentrations (44-370 GC/L) were relatively lower than the concentrations of HAdV (64-1,000 GC/L). Estimated mean adenovirus risks were greater for snorkelling than for swimming by roughly one to two orders of magnitude and estimated mean illness risks for snorkelling were >32/1,000. Human faecal contamination is frequent in the Yal-ku lagoon, which is associated with human gastrointestinal illness.

Epidemiology of Keratoconjunctivitis Across India from 2017 to 2019: A Multicentric Hospital-Based Study

ABSTRACT Purpose Conjunctivitis is one of the most common ocular conditions in clinical practice. Human adenoviruses have been the common causative agents known to cause epidemic kerato-conjunctivitis (EKC) in India from 1996 to 2019 with a positivity range of 13.8%–65.2%. The current study was initiated to throw light on the distribution of keratoconjunctivitis causing agents across India covering a span of 3 years. Methods A total of 709 swabs were collected from patients in viral transport medium (VTM), and real-time PCR was done to identify agents including Adenovirus (HAdV), Enterovirus, HSV, and Chlamydia. Results 47.8% of the samples were positive for HAdV followed by HSV (3.4%), Enterovirus (2.7%), and Chlamydia (0.6%). Overall, 386 people (54.4%) tested positive for one of these infections, with Chandigarh (88.4%) and Port Blair (71.7%) showing higher positivity rate. Pre-auricular lymphadenopathy and follicles were significantly associated with increased risk of conjunctivitis. Conclusion Epidemiology of keratoconjunctivitis in the current study revealed HAdV to be predominant causative agent. Knowledge gained in such epidemiological studies guide us in outbreak expectations, limit antibiotic over-prescription, and enhance disease prevention.

PDA-mediated colorimetric-fluorescence co-enhanced immunochromatography assay for the simultaneous and rapid detection of SARS-CoV-2 and HAdV

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and human adenovirus (HAdV) are highly contagious to humans and cause similar symptoms, with HAdV occurring mostly in children. Therefore, a flexible, sensitive, and accurate immunochromatographic assay (ICA) capable of recognizing SARS-CoV-2 and HAdV is urgently needed in the face of infected pediatric patients. Here, we developed a colorimetric-fluorescent co-enhanced dual-mode ICA sensor for the rapid and accurate detection of SARS-CoV-2 and HAdV. A multilayered composite nanomaterial named polydopamine@dual shell quantum dots nanobead (PDQB), which consisted of a polydopamine core (colorimetric) and a two-layered shell (fluorescent) integrated with thousands of small quantum dots, was prepared to replace conventional colorimetric nanotags (colloidal gold, AuNPs) and fluorescent nanotags (quantum nanobeads, QBs) while providing an easily readable colorimetric signal and an easily quantifiable fluorescent signal. The sensitivities of the PDQB-based ICA for SARS-CoV-2 and HAdV were 0.1 ng/mL and 104 copies/mL in colorimetric mode, respectively, and 0.005 ng/mL and 500 copies/mL in fluorescent mode, respectively. The limit of detection (LOD) of the PDQB-based ICA in colorimetric mode was 5 times lower than that of conventional AuNP-ICA. The LOD of the PDQB-based ICA in fluorescent mode was 10 times lower than that of commercial QB-ICA. This work provides a novel strategy for the detection of respiratory viruses, that realizes the co-enhancement and integration of colorimetric and fluorescent methods with high accuracy and convenience. Excitingly, the co-enhancement of colorimetric and fluorescent modes can enable the detection of respiratory viruses in different scenarios, thus guiding epidemic prevention and control.

Inhibition of adenovirus transport from the endosome to the cell nucleus by rotenone.

Regardless of the clinical impact of human adenovirus (HAdV) infections in the healthy population and its high morbidity in immunosuppressed patients, a specific treatment is still not yet available. In this study, we screened the CM1407 COST Action's chemical library, comprising 1,233 natural products to identify compounds that restrict HAdV infection. Among them, we identified rotenolone, a compound that significantly inhibited HAdV infection. Next, we selected four isoflavonoid-type compounds (e.g., rotenone, deguelin, millettone, and tephrosin), namely rotenoids, structurally related to rotenolone in order to evaluate and characterized in vitro their antiviral activities against HAdV and human cytomegalovirus (HCMV). Their IC50 values for HAdV ranged from 0.0039µM for rotenone to 0.07µM for tephrosin, with selective indices ranging from 164.1 for rotenone to 2,429.3 for deguelin. In addition, the inhibition of HCMV replication ranged from 50% to 92.1% at twice the IC50 concentrations obtained in the plaque assay for each compound against HAdV. Our results indicated that the mechanisms of action of rotenolone, deguelin, and tephrosin involve the late stages of the HAdV replication cycle. However, the antiviral mechanism of action of rotenone appears to involve the alteration of the microtubular polymerization, which prevents HAdV particles from reaching the nuclear membrane of the cell. These isoflavonoid-type compounds exert high antiviral activity against HAdV at nanomolar concentrations, and can be considered strong hit candidates for the development of a new class of broad-spectrum antiviral drugs.

Evaluation of a custom designed hybridisation assay for whole genome sequencing of human adenoviruses direct from clinical samples

Background: Human Adenoviruses are a common cause of disease and can cause significant morbidity and mortality in immunocompromised patients. Nosocomial transmission events can occur with whole genome sequencing playing a crucial role. This study evaluates the performance of a custom designed SureSelectXT target enrichment assay based on 14 adenovirus genomes for sequencing direct from clinical samples.Methods: Modifications were made to the SureSelectXT low input protocol to enhance performance for viral targets. Consensus sequences were generated using an in-house designed three stage bioinformatics pipeline. We assessed, percentage of on target reads, average depth of coverage and percentage genome coverage to determine assay performance across a range of sample matrices.Results: Whole genome sequences were successfully generated for 91.6 % of samples assessed. Adenovirus DNA concentration was a good indicator of enrichment success. Highly specific enrichment was observed with only 6 % of samples showing < 50 % on target reads. Respiratory and faecal samples performed well where bloods showed higher levels of non-specific enrichment likely confounded by low adenovirus DNA concentrations. Protocol performance did not appear impacted by Adenovirus type or species.Conclusion: Overall performance of this modified SureSelectXT protocol appears in line with previously published works although there are some confounding factors requiring further investigation. The use of a small RNA bait set has the potential to reduce associated costs which can be prohibitive.

Human conjunctiva organoids to study ocular surface homeostasis and disease

The conjunctival epithelium covering the eye contains two main cell types: mucus-producing goblet cells and water-secreting keratinocytes, which present mucins on their apical surface. Here, we describe long-term expanding organoids and air-liquid interface representing mouse and human conjunctiva. A single-cell RNA expression atlas of primary and cultured human conjunctiva reveals that keratinocytes express multiple antimicrobial peptides and identifies conjunctival tuft cells. IL-4/-13 exposure increases goblet and tuft cell differentiation and drastically modifies the conjunctiva secretome. Human NGFR+ basal cells are identified as bipotent conjunctiva stem cells. Conjunctival cultures can be infected by herpes simplex virus 1 (HSV1), human adenovirus 8 (hAdV8), and SARS-CoV-2. HSV1 infection was reversed by acyclovir addition, whereas hAdV8 infection, which lacks an approved drug therapy, was inhibited by cidofovir. We document transcriptional programs induced by HSV1 and hAdV8. Finally, conjunctival organoids can be transplanted. Together, human conjunctiva organoid cultures enable the study of conjunctival (patho)-physiology.

Molecular detection of Human Adenovirus and Human Bocavirus in wastewater and drain water in the western Nile Delta of Egypt

Abstract HAdV and HBoV are frequently found in different water sources throughout the year. Due to their health impact on humans, continuous monitoring of these viruses is considered an important tool to evaluate their circulation within society. This study investigates the prevalence of HAdV and HBoV in different environmental samples collected from the western Nile Delta of Egypt. In total, 264 samples were collected from treated and untreated sewage (48), sewage sludge (24), drainage water (96), and drainage sediment (96) from December 2019 to November 2021. Molecular detection was performed to detect both viruses in the collected samples. The results indicated that HAdV was more prevalent than HBoV in all types of sewage samples, with the highest detection rate in untreated sewage. Both viruses were relatively lower in sludge sewage samples, while in drainage water and sediment samples were detected with similar frequencies, although the detection rates were generally lower than those observed in sewage samples. Moreover, no significant seasonal distribution of both viruses was observed throughout the year. The results of this study emphasize the importance of monitoring the presence of these viruses in environmental samples with proper treatment and disposal of sewage to ensure public health and safety.

TNIK regulation of interferon signaling and endothelial cell response to virus infection.

Traf2 and Nck-interacting kinase (TNIK) is known for its regulatory role in various processes within cancer cells. However, its role within endothelial cells (ECs) has remained relatively unexplored. Leveraging RNA-seq data and Ingenuity Pathway Analysis (IPA), we probed the potential impact of TNIK depletion on ECs. Examination of RNA-seq data uncovered more than 450 Differentially Expressed Genes (DEGs) in TNIK-depleted ECs, displaying a fold change exceeding 2 with a false discovery rate (FDR) below 0.05. IPA analysis unveiled that TNIK depletion leads to the inhibition of the interferon (IFN) pathway [-log (p-value) >11], downregulation of IFN-related genes, and inhibition of Hypercytokinemia/Hyperchemokinemia [-log (p-value) >8]. The validation process encompassed qRT-PCR to evaluate mRNA expression of crucial IFN-related genes, immunoblotting to gauge STAT1 and STAT2 protein levels, and ELISA for the quantification of IFN and cytokine secretion in siTNIK-depleted ECs. These assessments consistently revealed substantial reductions upon TNIK depletion. When transducing HUVECs with replication incompetent E1-E4 deleted adenovirus expressing green fluorescent protein (Ad-GFP), it was demonstrated that TNIK depletion did not affect the uptake of Ad-GFP. Nonetheless, TNIK depletion induced cytopathic effects (CPE) in ECs transduced with wild-type human adenovirus serotype 5 (Ad-WT). Our findings suggest that TNIK plays a crucial role in regulating the EC response to virus infections through modulation of the IFN pathway.

SNP IFNG rs1861494 gene polymorphism in pediatric patients with gastroenteritis and its relation to co-infectious with Adenovirus and Cryptosporidium

Introduction and Aim: The most common cause of infantile gastroenteritis is enteric adenoviruses, while Cryptosporidiosis is caused by Cryptosporidium. The function of IFN-gamma (IFNG/IFN-y) in immunological responses, inflammation, and autoimmune disease is increasingly recognized. This study aimed to examine intestinal co-infections of Cryptosporidium and adenovirus in pediatrics and the role of IFNG rs1861494 polymorphism to gastroenteritis among Iraqi children. Materials and Methods: This case control study included 75 pediatric patients with severe gastroenteritis, aged 3-120 months, with a mean age of 30.64 months. The apparently healthy control (AHC) in this study included 25 pediatric individuals with a mean age of 27.64 months. Total genomic DNA extracted from stool specimens were subjected to PCR detection of human adenovirus virus. SNP IFNG rs1861494 polymorphism was detected using the ARMS-PCR technique. Results: The percentage of males to female ratio in this study was 54:46. The positive PCR result of HADV7 was 28% (14 of 50 cases) while the positive result of Cryptosporidium was 6% (6 of 75 cases).Genotyping studies showed the GG genotype increased at rate OR=2.67 as compared to AG and AA genotypes for the SNP IFNG rs1861494. The SNP IFNG rs1861494 results in gastroenteritis were highly correlated with HADV7 and Cryptosporidium (respectively: r = 0.968, P = 0.007 as well as r = 0.984, P = 0.008). Conclusions: Adenovirus diarrhea in Iraqi pediatric patients is associated with SNP rs1861494 polymorphism and could possibly be used as a biomarker in assessing the susceptibility of Iraqi children to the virus.

Oral USC-093, a novel homoserinamide analogue of the tyrosinamide (S)-HPMPA prodrug USC-087 has decreased nephrotoxicity while maintaining antiviral efficacy against human adenovirus infection of Syrian hamsters

Adenovirus infections of immunocompromised humans are a significant source of morbidity and mortality. Presently, there is no drug specifically approved for the treatment of adenovirus infections by the FDA. The state-of-the-art treatment of such infections is the off-label use of cidofovir, an acyclic nucleotide phosphonate. While cidofovir inhibits adenovirus replication, it has dose-limiting kidney toxicity. There is an apparent need for a better compound to treat adenovirus infections. To this end, we have been developing acyclic nucleotide phosphonate prodrugs that utilize an amino acid scaffold equipped with a lipophilic modifier. Here, we compare the antiviral potential of two prodrugs of HPMPA that differ only in the amino acid-based promoiety: USC-087, based on an N-hexadecyl tyrosinamide, and USC-093, based on an N-hexadecyl serinamide. Oral administration of both compounds was very efficacious against disseminated HAdV-C6 infection in immunosuppressed Syrian hamsters, suppressing virus replication and mitigating pathology even when treatment was withheld until 4 days after challenge. We saw only marginal efficacy after respiratory infection of hamsters, which may reflect suboptimal distribution to the lung. Importantly, neither compound induced intestinal toxicity, which was observed as the major adverse effect in clinical trials of brincidofovir, a prodrug of cidofovir which also contains a C-16 modifier. Notably, we found that there was a significant difference in the nephrotoxicity of the two compounds: USC-087 caused significant kidney toxicity while USC-093 did not, at effective doses. These findings will be valuable guidepoints in the future evolution of this new class of potential prodrugs to treat adenovirus infections.

Rapid detection of human adenovirus subgroup B using recombinase polymerase amplification assay.

Human adenovirus subgroup B (HAdV B) is one of the major pathogens of human respiratory virus infections, which has considerable transmission and morbidity in a variety of populations. Therefore, rapid and specific detection of HAdV B in clinical samples is essential for diagnosis. This study aimed to develop a product for rapid nucleic acid detection of HAdV B using recombinase polymerase amplification assay (RPA) and validate the performance of this method by using clinical samples. Results showed that this method achieved a lower limit of detection (LOD) of 10 copies/μL and had no cross-reactivity with other adenovirus subgroups or respiratory pathogens. In addition to high sensitivity, it can be completed within 30min at 40°C. There is no need to perform nucleic acid extraction on clinical samples. Taking qPCR as the gold standard, the RPA assay possessed a high concordance (Cohen's kappa, 0.896; 95% CI 0.808-0.984; P < 0.001), with a sensitivity of 87.80% and a specificity of 100.00%. The RPA assay developed in this study provided a simple and highly specific method, making it an important tool for rapid adenovirus nucleic acid detection and facilitating large-scale population screening in resource-limited settings.

NEAR INFRARED-RESPONSIVE HYDROGELS CONTAINING ADENOVIRAL VECTORS: APPLICATION IN BONE REGENERATION

We have developed plasmonic fibrin-based hydrogels that incorporate gold nanoparticles which transduce incident near-infrared (NIR) light into heat. Human adenovirus serotype type-5 vectors encoding a firefly luciferase (fLuc) coding sequence driven by a heat-inducible promoter were incorporated into the hydrogels. Transmission electronic microscopic analysis revealed that the adenoviral vectors were associated to the fibrin fibers. In vitro experiments in which human cells were cultured with plasmonic hydrogels showed that the adenoviral vectors can diffuse from the hydrogels, transduce the cells, and stimulate heat-induced transgene expression upon NIR irradiation. The hydrogels were implanted in 4.2 mm drill hole defects generated in the humerus of male rabbits. Three days after implantation, the defects were NIR-irradiated. Six h later, the animals were euthanized and samples from the bone defect zone were processed for immunohistochemical analyses using a specific fLuc antibody. The results showed strong expression of fLuc in tissues surrounding the implants of NIR-irradiated rabbits, while non- irradiated animals exhibited negligible expression. We next aimed to use the temperature increase to induce the production of transgenic bone morphogenetic protein 6 (BMP-6), using safe gene switches that can provide tighter control of in vivo transgene expression than heat-inducible promoters. These switches are only activated by heat in the presence of rapamycin and maintain a high level of targeted transgene expression for several days after heat activation. Adenoviral vectors encoding the safe switches that control the expression of BMP-6 were incorporated to the composites. The resulting NIR-responsive hydrogels were implanted in the bone defects generated in rabbits and used as a platform to transduce host cells, generate local hyperthermia and stimulate BMP-6 production.Acknowledgements: This research was supported by grants RTI2018-095159-B-I00 and PID2021-126325OB-I00 (MCIN/AEI/10.13039/501100011033 and “ERDF A way of making Europe”), by grant P2022/BMD- 7406 (Regional Government of Madrid). M.A.L-J. is the recipient of predoctoral fellowship PRE2019-090430 (MCIN/AEI/10.13039/501100011033).

Severe lower respiratory tract infections are associated with human adenovirus in hospitalised children in a high HIV prevalence area.

Viral causes of lower respiratory tract infections (LRTIs) are associated with increased mortality in children aged <5 years (U5). Human adenovirus (HAdV) has been associated with severe LRTI; however, its relationship with HIV and malnutrition in South Africa (SA) is not understood. To identify the prevalence of and factors associated with HAdV LRTIs in hospitalised U5 childen. Clinical and viral data on U5 children hospitalised with severe LRTI from January 2018 to June 2020 at King Edward VIII Hospital, Durban, SA, including results of a multiplex polymerase chain reaction (PCR) panel assay for respiratory viruses, were retrieved from inpatient files and laboratory databases and retrospectively analysed. Standard descriptive statistics and Pearson's χ², Fisher's exact and Mann-Whitney tests were used to determine significant associations with HAdV LRTI. Among the 206 viral assays analysed (15.6% of all LRTI admissions), HAdV was the most common virus identified. The cohort had a median (interquartile range) age of 5 (2 - 13) months, 47.3% had perinatal HIV exposure, and 34.5% had severe acute malnutrition (SAM). No seasonal pattern with HAdV could be demonstrated. SAM and prematurity were significant risk factors for readmission, and perinatal HIV exposure was a significant risk factor for presence of multiple viruses on analysis of a respiratory specimen. Detection of HAdV was not associated with an increased risk of requiring oxygen or ventilatory support. HAdV was the most common virus found on analysis of multiplex PCR panel results in children hospitalised with severe LRTI in SA, where high rates of HIV exposure may result in increased susceptibility to viral co-infections. The role of HAdV as a cause of severe LRTI in SA infants, who have high rates of HIV exposure, requires greater scrutiny. What the study adds. This study provides retrospective data identifying human adenovirus (HAdV) as the most common cause of severe lower respiratory tract infection (LRTI) in children aged <5 years (U5). The impact of respiratory syncytial virus as a common pathogen in children is well established. The study confirms anecdotal evidence that HAdV is an important disease-causing pathogen associated with LRTI. Children with perinatal HIV exposure and severe acute malnutrition (SAM) may be particularly susceptible.Implications of the findings. HAdV must be considered a major cause of severe LRTI in U5 children. Children with LRTI who had perinatal HIV exposure and those with SAM need to be tested for HAdV and to be monitored for severe disease.

Epidemiology and molecular detection of human adenovirus and non-polio enterovirus in fecal samples of children with acute gastroenteritis: A five-year surveillance in northern Brazil.

Acute gastroenteritis (AGE) is a common pediatric infection that remains a significant cause of childhood morbidity and mortality worldwide, especially in low-income regions. Thus, the objective of this study was to detect human adenovirus (HAdV) and non-polio enterovirus (NPEV) in fecal samples from the Gastroenteritis Surveillance Network, and to identify circulating strains by nucleotide sequencing. A total of 801 fecal samples were tested using qPCR/RT-qPCR, and 657 (82.0%) were inoculated into HEp-2C and RD cell lines. The HAdV and NPEV positivity rates obtained using qPCR/RT-qPCR were 31.7% (254/801) and 10.5% (84/801), respectively, with 5.4% (43/801) co-detection. Cytopathic effect was observed in 9.6% (63/657) of patients, 2.7% (18/657) associated with HAdV, and 6.2% (41/657) associated with NPEV after testing by ICC-PCR. A comparison of the two methodologies demonstrated an agreement of 93.5% for EVNP and 64.4% for HAdV. These two viruses were detected throughout the study period, with HAdV positivity rates ranging from 41% in Amapá to 18% in Pará. The NEPV varied from 18% in Pará/Rondônia to 3% in Acre. The most affected age group was over 60 months for both HAdV and NPEV. Samples previously positive for rotavirus and norovirus, which did not show a major difference in the presence or absence of diarrhea, fever, and vomiting, were excluded from the clinical analyses of these two viruses. These viruses circulated over five years, with a few months of absence, mainly during the months corresponding to the waves of SARS-CoV-2 infection in Brazil. Five HAdV species were identified (A, B, C, D, and F), with a greater predominance of HAdV-F41 (56.5%) followed by HAdV-C (15.2%). Three NPEV species (A, B, and C) were detected, with serotypes E14 (19.3%) and CVA-24 (16.1%) being the most prevalent. The present study revealed a high diversity of NPEV and HAdV types circulating in children with AGE symptoms in the northern region of Brazil.

Genotypes and Phylogenetic Analysis of Human Adenovirus in Hospitalized Pneumonia and Influenza-Like Illness Patients in Jiangsu Province, China (2013-2021).

Human adenovirus (HAdV) is common pathogens that cause various respiratory diseases. The genetic diversity of viruses caused by recombination is considered to be the main source of emerging outbreaks. The aim of this study is to explore the evolutionary relationship and recombination events of HAdV genome in respiratory tract infections in Jiangsu Province. Whole-genome sequencing (WGS) technology was used to sequence 66 patients with HAdV infection (37 patients with influenza-like illness (ILI) and 29 hospitalized patients with pneumonia) from Jiangsu Province. Epidemiological analysis was performed on hospitalized pneumonia and ILI patients infected with HAdV. Subsequently, phylogenetic, recombination, and nucleotide and amino acid identity analyses were performed. Epidemiological analysis of patients undergoing WGS showed that 75.7% of ILI patients were infected with the HAdVB strain and 69.0% of hospitalized pneumonia patients were infected with the HAdVC strain. Moreover, the hospitalized pneumonia and ILI patients infected with HAdV were different in region and time. The strains of HAdVB3 and HAdVB7 genotypes were mainly infected in 2015 and 2017, and the strains of HAdVC1 and HAdVC2 genotypes were mainly infected in 2020. The results of histogram analysis showed that the HAdV strain mainly infected children under 5 years old. In addition, 36 novel recombinant strains were identified. The discovery of these recombinant strains may contribute to understanding the epidemiology of HAdV and research on related vaccines. Furthermore, the percentage of nucleotide and amino acid identities revealed a high level of genetic conservation within isolates from HAdVB3, HAdVB7, HAdVC1, HAdVC2 and HAdVC5 genotypes. The WGS analysis reveals the evolutionary relationships and recombination events of HAdV strains in Jiangsu Province, which is helpful to deepen the understanding of HAdV epidemiology and evolution. In addition, it provides a basis for the formulation of public health strategies in Jiangsu Province.

Detection of Human Adenovirus from Clinical Samples of Conjunctivitis by Real-time PCR: A Cross-sectional Descriptive Study

Introduction: Human Adenoviruses (HAdV) have been implicated in a variety of infections, including conjunctivitis, respiratory tract infections, genitourinary infections and gastroenteritis. Epidemic Keratoconjunctivitis (EKC) is a severe ocular surface infection strongly associated with HAdV, known to occur in widespread outbreaks. HAdV species B, D and E are associated with ocular manifestations ranging from simple follicular conjunctivitis (types 3, 4, 7) and pharyngoconjunctival fever (types 3, 7, 14) to the more severe EKC (types 8, 19, 37, 53, 54, 56 and 64). Viral cell cultures of conjunctival specimens help confirm adenovirus infection through the characteristic Cytopathic Effect (CPE) and Polymerase Chain Reaction (PCR) is the best standard method to diagnose viral conjunctivitis due to its sensitivity, accuracy and rapidity. Aim: To diagnose HAdV in clinically suspected cases of viral conjunctivitis using real-time PCR. To isolate HAdV in viral cell culture and confirm it by observing its characteristic CPE and performing adenoviral real-time PCR. Materials and Methods: A cross-sectional descriptive study was conducted from July 2023 to September 2023 at the state-level Virus Research and Diagnostic Laboratory (VRDL) at Bangalore Medical College and Research Institute (BMCRI), Bengaluru, Karnataka, India. A total of 45 conjunctival swab samples from patients with suspected viral conjunctivitis, who attended the Ophthalmology Outpatient Department (OPD) or were admitted to the wards, were included in the study. Conjunctival swab samples collected in Viral Transport Medium (VTM) were centrifuged at 3000 rpm for five minutes at 4°C. Real-time PCR was performed and samples positive for HAdV by real-time PCR were taken up for viral cell culture. Confirmatory PCR was conducted for samples showing CPEs in the cell line. The data collected were analysed using descriptive statistics. Results: The majority of patients in the present study were in the age group of 18-45 years, comprising 25 (56%) of the total patients. Out of 45 conjunctival samples tested, 8 samples were positive in adenovirus real-time PCR. All eight PCR-positive samples showed a CPE in viral cell culture on the A549 cell line. The study found PCR positivity in 8 samples (17.7%), while adenoviral recovery from cell culture was observed in 6 samples (13.3%). Conclusion: Real-time PCR has become the standard diagnostic procedure for detecting adenovirus conjunctivitis. Rapid and accurate diagnosis is key to interrupting the contagious spread of adenoviral conjunctivitis, along with timely treatment.

Severe Adenovirus Infections in Young Adults

&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Despite traditionally affecting individuals with compromised immune systems, severe human adenoviruses (HAdV) infections have been observed in young adults without immunodeficiency over recent decades.&lt;br /&gt;&lt;strong&gt;Case Presentation:&lt;/strong&gt; This case study documents two young patients infected with HAdV who experienced multi-organ failure,&lt;br /&gt;manifesting as respiratory distress, vasoplegia, myocardial injury, and kidney impairment. The resolution of organ failure&lt;br /&gt;correlated with the clearance of viral load, suggesting a potential prognostic indicator for infection outcomes. While the first&lt;br /&gt;case demonstrated a positive outcome with the resolution of viremia, regrettably, the second case succumbed to myocardial&lt;br /&gt;infarction followed by cardiac arrest. The use of cardiac monitoring may aid in the identification of significant myocardial&lt;br /&gt;complications associated with HAdV. Although various treatments, including the antiviral CIDOFOVIR, have been reported for&lt;br /&gt;immunocompromised individuals, it was not administered to our patients due to the favorable trajectory of the first case.&lt;br /&gt;Tragically, the second case passed away before any potential antiviral intervention could be initiated.&lt;br /&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;HAdV infections can cause severe acute respiratory distress syndrome with multi-organ failure including severe&lt;br /&gt;cardiac complications in young non-immunocompromised adults.&lt;/p&gt;