Adenovirus Serotype Research Articles

Heterologous saRNA Prime, DNA Dual-Antigen Boost SARS-CoV-2 Vaccination Elicits Robust Cellular Immunogenicity and Cross-Variant Neutralizing Antibodies.

We assessed if immune responses are enhanced in CD-1 mice by heterologous vaccination with two different nucleic acid-based COVID-19 vaccines: a next-generation human adenovirus serotype 5 (hAd5)-vectored dual-antigen spike (S) and nucleocapsid (N) vaccine (AdS+N) and a self-amplifying and -adjuvanted S RNA vaccine (AAHI-SC2) delivered by a nanostructured lipid carrier. The AdS+N vaccine encodes S modified with a fusion motif to increase cell-surface expression and an N antigen modified with an Enhanced T-cell Stimulation Domain (N-ETSD) to direct N to the endosomal/lysosomal compartment and increase MHC class I and II stimulation potential. The S sequence in the AAHI-SC2 vaccine comprises the D614G mutation, two prolines to stabilize S in the prefusion conformation, and 3 glutamines in the furin cleavage region to confer protease resistance. CD-1 mice received vaccination by homologous and heterologous prime > boost combinations. Humoral responses to S were the highest with any regimen that included the AAHI-SC2 vaccine, and IgG bound to wild type and Delta (B.1.617.2) variant S1 at similar levels. An AAHI-SC2 prime followed by an AdS+N boost particularly enhanced CD4+ and CD8+ T-cell responses to both wild type and Delta S peptides relative to all other vaccine regimens. Sera from mice receiving AAHI-SC2 homologous or heterologous vaccination were found to be highly neutralizing for all pseudovirus strains tested: Wuhan, Beta, Delta, and Omicron strains. The findings here, taken in consideration with the availability of both vaccines in thermostable formulations, support the testing of heterologous vaccination by an AAHI-SC2 > AdS+N regimen in animal models of SARS-CoV-2 infection to assess its potential to provide increased protection against emerging SARS-CoV-2 variants particularly in regions of the world where the need for cold-chain storage has limited the distribution of other vaccines.

P-624 The type of vaccine received against SARS-CoV-2 does not affect ovarian function in an Assisted Reproduction cycle

Study questionDo the different types of vaccines against SARS-CoV-2 influence the results of an Assisted Reproduction treatment?Summary answerThe type of vaccine administered against SARS-CoV-2 does not affect the results in women performing an Assisted Reproduction treatment.What is known alreadySince the COVID-19 pandemic was declared, the search for vaccines has become the priority, so its development has represented a step towards herd immunity in a short period of time. Despite this encouraging advance, vaccine hesitancy in reproductive-aged women has been heightened because of the spread of misinformation stating that COVID-19 vaccines will cause sterility. Due to the lack of information and the clinical relevance, the objective of this work was to evaluate the impact of the different types of vaccines on women’s fertility.Study design, size, durationRetrospective and observational study during January-October 2021 in women vaccinated against SARS-CoV-2 and performing an Assisted Reproductive treatment in any of the 11 clinics belonging to the IVIRMA group in Spain. The Control group included patients performing a treatment during the same study period but who had not yet been vaccinated. The study was approved by an Institutional Review Board (2109-MADR-084-AR)Participants/materials, setting, methodsAll women, those who had received the complete vaccination schedule, regardless of the type of vaccine administered mRNA or viral vector, and women from the control group underwent the same ovarian stimulation protocol. The Assisted Reproduction treatment was performed with their own oocytes in all cases. Statistical analyses were performed using the Statistical Package for Social Sciences 19.0 (IBM Corporation, Armonk, NY, USA).Main results and the role of chanceWe included 510 patients distributed as follows: 13.5% (n = 69) received a viral vector vaccine either the adenovirus serotype 26 vector vaccine (Ad26.CoV2.S; Johnson & Johnson; n = 31) or the chimpanzee adenovirus vector vaccine (ChAdOx; AstraZeneca; n = 38). The remaining 86.5% (n = 441) received an mRNA vaccine from either Pfizer-BioNTech (n = 336) or Moderna (n = 105). Sample size for control group was n = 1190Our results showed that women vaccinated with Johnson & Johnson have a higher average age (39.7 ± 4.3) than the other groups, although no statistical difference was observed (p = 0.072); that is, AstraZeneca (36.8 ± 1.7), Moderna (35.7 ± 1.5), Pfizer (34.6 ± 1.6) and the control group (37.8 ± 2.7). This circumstance did not affect other parameters such as the days of stimulation (p = 0.336) or the doses of FSH administered (p = 0.392), where no statistical differences were recorded between the vaccinated and the control group. Finally, the number of oocytes were as follows, Johnson & Johnson (9.2 ± 2.6), AstraZeneca (7.7 ± 1.2), Moderna (11.3 ± 1.8), Pfizer (12.6 ± 1.0), and the control group (10.2 ± 1.5), p = 0.06.Limitations, reasons for cautionThis is an observational study, and thus possible confounders cannot be excluded entirely. More data are needed to draw firm conclusions, and it will be critical to increase the sample size to check if the results observed in this work remains in the general populationWider implications of the findingsThis is the first study to assess whether the type of vaccine administered against SARS-CoV-2, mRNA, or viral vector, affects ovarian function in ART. These early findings suggest no measurable detrimental on ovarian response regardless of vaccine received.Trial registration numbernot applicable

Probiotics Surface-Delivering Fiber2 Protein of Fowl Adenovirus 4 Stimulate Protective Immunity Against Hepatitis-Hydropericardium Syndrome in Chickens.

Background and ObjectivesHepatitis-hydropericardium syndrome (HHS) caused by Fowl adenoviruses serotype 4 (FAdV-4) leads to severe economic losses to the poultry industry. Although various vaccines are available, vaccines that effectively stimulate intestinal mucosal immunity are still deficient. In the present study, novel probiotics that surface-deliver Fiber2 protein, the major virulence determiner and efficient immunogen for FAdV-4, were explored to prevent this fecal–oral-transmitted virus, and the induced protective immunity was evaluated after oral immunization.MethodsThe probiotic Enterococcus faecalis strain MDXEF-1 and Lactococcus lactis NZ9000 were used as host strains to deliver surface-anchoring Fiber2 protein of FAdV-4. Then the constructed live recombinant bacteria were orally vaccinated thrice with chickens at intervals of 2 weeks. Following each immunization, immunoglobulin G (IgG) in sera, secretory immunoglobulin A (sIgA) in jejunum lavage, immune-related cytokines, and T-cell proliferation were detected. Following challenge with the highly virulent FAdV-4, the protective effects of the probiotics surface-delivering Fiber2 protein were evaluated by verifying inflammatory factors, viral load, liver function, and survival rate.ResultsThe results demonstrated that probiotics surface-delivering Fiber2 protein stimulated humoral and intestinal mucosal immune responses in chickens, shown by high levels of sIgA and IgG antibodies, substantial rise in mRNA levels of cytokines, increased proliferative ability of T cells in peripheral blood, improved liver function, and reduced viral load in liver. Accordingly, adequate protection against homologous challenges and a significant increase in the overall survival rate were observed. Notably, chickens orally immunized with E. faecalis/DCpep-Fiber2-CWA were completely protected from the FAdV-4 challenge, which is better than L. lactis/DCpep-Fiber2-CWA.ConclusionThe recombinant probiotics surface-expressing Fiber2 protein could evoke remarkable humoral and cellular immune responses, relieve injury, and functionally damage target organs. The current study indicates a promising method used for preventing FAdV-4 infection in chickens.

Acute Hepatitis of Unknown Origin in Children: Early Observations from the 2022 Outbreak.

Recent reports of acute hepatitis of unknown origin in previously healthy children have been increasing worldwide. The main characteristics of the affected children were jaundice and gastrointestinal symptoms. Their serum aminotransaminase levels were above 500 IU/L, with negative tests for hepatitis viruses A–E. By 31 May 2022, the outbreak had affected over 800 children under the age of 16 years in more than 40 countries, resulting in acute liver failure in approximately 10%, including at least 21 deaths and 38 patients requiring liver transplantation. There was still no confirmed cause or causes, although there were several different working hypotheses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), adenovirus serotype 41, or SARS-CoV-2 superantigen-mediated immune cell activation. Here, we review early observations of the 2022 outbreak which may inform diagnosis, treatment, and prevention in the context of an overlapping COVID-19 pandemic.

Abstract 3551: Oncolytic adenovirus serotype 35-mediated activation of NK cells leads to efficient anti-tumor effects

Abstract Oncolytic adenoviruses (OAds) are getting much attention as cancer immunotherapy agents. Almost all OAds are based on human adenovirus serotype 5 (Ad5) (OAd5). However, OAd5 has several drawbacks which possibly attenuate the anti-tumor effects of OAd5. More than 80% of adults have neutralizing antibodies against Ad5. The expression of coxsackievirus-adenovirus receptor (CAR), which is the primary infection receptor of Ad5, often declines on malignant tumor cells. Previously, we have developed a novel OAd fully composed of human adenovirus serotype 35 (Ad35) (OAd35) (Ono et al., Mol.Ther.Oncolytics, 2021). Only 20% or fewer adults have neutralizing antibodies against Ad35. Ad35 recognizes CD46, which is highly expressed on malignant tumor cells, as an infection receptor. OAd35 showed efficient tumor cell lysis activities in many types of tumor cells; however, the mechanism of anti-tumor effects of OAd35 remains to be fully evaluated. In this study, we examined the mechanism of anti-tumor effects of OAd35. Although OAd35 showed efficient tumor growth suppression at levels comparable to OAd5 following intratumoral administration in nude mice bearing H1299 human lung cancer xenografts, the genome copy numbers of OAd35 in the tumor were about 500-folds lower than those of OAd5 at 18 days after administration and were almost background levels. These data suggest that other mechanisms different from infection of tumor cells with OAd35 were involved in the anti-tumor effects of OAd35. Since previous studies reported that Ad35 efficiently activated innate immunity in natural killer (NK) cells, we examined whether OAd35-mediated activation of NK cells contributed to the anti-tumor effects of OAd35. Intratumoral administration of OAd35 resulted in efficient infiltration and activation of NK cells in H1299 tumors. In contrast, OAd5 did not apparently induce the infiltration and activation of NK cells in the tumors. OAd35-mediated activation and infiltration of NK cells were also found in B16 tumor-bearing immunocompetent mice. Next, in order to examine the roles of NK cells in the anti-tumor effects of OAd35, NK cells were depleted by intravenous injection of anti-GM1 antibody in nude mice, followed by intratumoral administration of OAd35. The depletion of NK cells significantly attenuated the anti-tumor effects of OAd35 on H1299 tumors. Furthermore, pretreatment with anti-GM1 antibody significantly increased the genome copy numbers of OAd35 in the tumor, suggesting that activated NK cells removed the OAd35-infected tumor cells. This study demonstrated that OAd35 significantly activated NK cells following intratumoral administration and that activated NK cells efficiently removed the tumor cells, leading to efficient tumor growth suppression. Citation Format: Ryosuke Ono, Kosuke Takayama, Fuminori Sakurai, Hiroyuki Mizuguchi. Oncolytic adenovirus serotype 35-mediated activation of NK cells leads to efficient anti-tumor effects [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3551.

Abstract LB097: A simian challenge model to evaluate the CD8-positive T cell response with cancer therapeutics

Abstract The demonstration that antibodies can be used to treat cancers has revolutionized the field of cancer immunotherapy and gave hope to many patients. Originally targeting tumor-specific antigen, new therapeutic antibodies target now immune checkpoint markers expressed on T cells. The advantage is to generate an active and long-lasting antitumor immunity with the achievement of a broad and polyclonal antitumor immunity, addressing better the heterogeneity of cancers and reducing the chances of immune escape. The development of such highly specific molecules is particularly challenging however when it comes to preclinical safety assessment and efficacy studies. An ideal animal model needs to possess the target so that the drug product is pharmacologically active. Moreover, such model should ideally provide further insight in determining the pharmacological action of the product and the determination of safety to maximize the usefulness in animal studies. Intriguingly, no equivalent of the traditional T-cell-dependent antibody response (TDAR) assay, which evaluates mainly the CD4+ T cells response, is currently available for the CD8+ T cell response. The goal of this study is to fill this gap with a Non-Human Primate (NHP) challenge model that will elicit a potent CD8+ T cell response. MHC I-genotyped Mauritian cynomolgus macaques (MCMs) were immunized with 3 replication incompetent recombinant adenovirus serotype 5 (Ad5) vectors encoding Gag, Nef or Pol SIV proteins. MCMs were distributed into 3 groups and received two intramuscular injections of adenoviruses 4 weeks apart. One group received the anti-PD-L1 Atezolizumab and a second received a molecule in early development targeting another immune checkpoint molecule. T cell activation was monitored with blood samples taken on a weekly basis for 8 weeks. Blood samples were used to characterize the T cell immunophenotypes and to correlate them with some functional assays. Vaccination was well tolerated and could be conducted on standard safety toxicology studies without compromising other endpoints. Adv5-SIVs modulated total T cells 2 weeks after a single vaccination, induced T-cell activation, T-cell proliferation and the expression of several checkpoint molecules. Moreover, functional assays confirmed that T-cell activation profile correlated with an antigen-specific CD8 T-cell response. Compounds administered differently enhanced the profile of T-cell activation, the upregulation of immune checkpoint molecules and antigen specific responses of their T cells. Moreover, both compounds extended the cytolytic activity of the antigen-specific CD8+ T cell up to 8 weeks. Taken together, the vaccination model we developed seems appropriate to study the pharmacology of new protein-based therapeutics targeting CD8+ T cells. Citation Format: Richard Graveline, Morad Haida, Carolyne Dumont, Dominic Poulin, Florence Poitout-Belissent, Rana Samadfan, Sven Kronenberg, Franziska Regenass-Lechner, Rodney Prell, Marie-Soleil Piche. A simian challenge model to evaluate the CD8-positive T cell response with cancer therapeutics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB097.

Transcriptome Analysis of Genes Responding to Infection of Leghorn Male Hepatocellular Cells With Fowl Adenovirus Serotype 4.

Fowl adenovirus serotype 4 (FAdV-4) is a highly pathogenic virus with a broad host range that causes huge economic losses for the poultry industry worldwide. RNA sequencing has provided valuable and important mechanistic clues regarding FAdV-4–host interactions. However, the pathogenic mechanism and host's responses after FAdV-4 infection remains limited. In this study, we used transcriptome analysis to identify dynamic changes in differentially expressed genes (DEGs) at five characteristic stages (12, 24, 36, 48, and 60 h) post infection (hpi) with FAdV-4. A total of 8,242 DEGs were identified based on comparison of five infection stages: 0 and 12, 12 and 24, 24 and 36, 36 and 48, and 48 and 60 hpi. In addition, at these five important time points, we found 37 common upregulated or downregulated DEGs, suggesting a common role for these genes in host response to viral infection. The predicted function of these DEGs using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that these DEGs were associated with viral invasion, host metabolic pathways and host immunosuppression. Interestingly, genes involved in viral invasion, probably EGR1, SOCS3, and THBS1, were related to FAdV-4 infection. Validation of nine randomly selected DEGs using quantitative reverse-transcription PCR produced results that were highly consistent with those of RNA sequencing. This transcriptomic profiling provides valuable information for investigating the molecular mechanisms underlying host–FAdV-4 interactions. These data support the current molecular knowledge regarding FAdV-4 infection and chicken defense mechanisms.

Vaccines based on the replication-deficient simian adenoviral vector ChAdOx1: Standardized template with key considerations for a risk/benefit assessment

Replication-deficient adenoviral vectors have been under investigation as a platform technology for vaccine development for several years and have recently been successfully deployed as an effective COVID-19 counter measure. A replication-deficient adenoviral vector based on the simian adenovirus type Y25 and named ChAdOx1 has been evaluated in several clinical trials since 2012.The Brighton Collaboration Benefit-Risk Assessment of VAccines by TechnolOgy (BRAVATO) was formed to evaluate the safety and other key features of new platform technology vaccines. This manuscript reviews key features of the ChAdOx1-vectored vaccines.The simian adenovirus Y25 was chosen as a strategy to circumvent pre-existing immunity to common human adenovirus serotypes which could impair immune responses induced by adenoviral vectored vaccines. Deletion of the E1 gene renders the ChAdOx1 vector replication incompetent and further genetic engineering of the E3 and E4 genes allows for increased insertional capability and optimizes vaccine manufacturing processes. ChAdOx1 vectored vaccines can be manufactured in E1 complementing cell lines at scale and are thermostable. The first ChAdOx1 vectored vaccines approved for human use, against SARS-CoV-2, received emergency use authorization in the UK on 30th December 2020, and is now approved in more than 180 countries.Safety data were compiled from phase I-III clinical trials of ChAdOx1 vectored vaccines expressing different antigens (influenza, tuberculosis, malaria, meningococcal B, prostate cancer, MERS-CoV, Chikungunya, Zika and SARS-CoV-2), conducted by the University of Oxford, as well as post marketing surveillance data for the COVID-19 Oxford-AstraZeneca vaccine. Overall, ChAdOx1 vectored vaccines have been well tolerated. Very rarely, thrombosis with thrombocytopenia syndrome (TTS), capillary leak syndrome (CLS), immune thrombocytopenia (ITP), and Guillain-Barre syndrome (GBS) have been reported following mass administration of the COVID-19 Oxford-AstraZeneca vaccine. The benefits of this COVID-19 vaccination have outweighed the risks of serious adverse events in most settings, especially with mitigation of risks when possible.Extensive immunogenicity clinical evaluation of ChAdOx1 vectored vaccines reveal strong, durable humoral and cellular immune responses to date; studies to refine the COVID-19 protection (e.g., via homologous/heterologous booster, fractional dose) are also underway.New prophylactic and therapeutic vaccines based on the ChAdOx1 vector are currently undergoing pre-clinical and clinical assessment, including vaccines against viral hemorrhagic fevers, Nipah virus, HIV, Hepatitis B, amongst others.

Structure-Morphology-Antimicrobial and Antiviral Activity Relationship in Silver-Containing Nanocomposites Based on Polylactide.

Green synthesis of silver-containing nanocomposites based on polylactide (PLA) was carried out in two ways. With the use of green tea extract, Ag+ ions were reduced to silver nanoparticles with their subsequent introduction into the PLA (mechanical method) and Ag+ ions were reduced in the polymer matrix of PLA-AgPalmitate (PLA-AgPalm) (in situ method). Structure, morphology and thermophysical properties of nanocomposites PLA-Ag were studied by FTIR spectroscopy, wide-angle X-ray scattering (WAXS), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC) methods. The antimicrobial, antiviral, and cytotoxic properties were studied as well. It was found that the mechanical method provides the average size of silver nanoparticles in the PLA of about 16 nm, while in the formation of samples by the in situ method their average size was 3.7 nm. The strong influence of smaller silver nanoparticles (3.7 nm) on the properties of nanocomposites was revealed, as with increasing nanosilver concentration the heat resistance and glass transition temperature of the samples decreases, while the influence of larger particles (16 nm) on these parameters was not detected. It was shown that silver-containing nanocomposites formed in situ demonstrate antimicrobial activity against gram-positive bacterium S. aureus, gram-negative bacteria E. coli, P. aeruginosa, and the fungal pathogen of C. albicans, and the activity of the samples increases with increasing nanoparticle concentration. Silver-containing nanocomposites formed by the mechanical method have not shown antimicrobial activity. The relative antiviral activity of nanocomposites obtained by two methods against influenza A virus, and adenovirus serotype 2 was also revealed. The obtained nanocomposites were not-cytotoxic, and they did not inhibit the viability of MDCK or Hep-2 cell cultures.

Oral immunization of recombinant Saccharomyces cerevisiae expressing fiber-2 of fowl adenovirus serotype 4 induces protective immunity against homologous infection

Hydropericardium-hepatitis syndrome (HHS) caused by fowl adenovirus (FAdV) serotype 4 strains is a highly contagious disease that causes significant economic loss to the global poultry industry. However, subunit vaccine against FAdV-4 infection is not yet commercially available to date. This study aims to explore the potential for oral immunization of recombinant Saccharomyces cerevisiae expressing Fiber-2 of FAdV-4 as a subunit vaccine. Here, we constructed recombinant S. cerevisiae (ST1814G/Fiber-2) expressing recombinant Fiber-2 (rFiber-2), which was displayed on the cell surface. To evaluate the immune response and protective effect of live recombinant S. cerevisiae, chickens were orally immunized with the constructed live ST1814G/Fiber-2, three times at 5-day intervals, and then challenged with FAdV-4. The results showed that oral administration of live ST1814G/Fiber-2 could stimulate the production of humoral immunity, enhance the body's antiviral activity and immune regulation ability, improve the composition of gut microbiota, provide protection against FAdV-4 challenge, reduce viral load in the liver, and alleviate the pathological damage of heart, liver, and spleen for chicken. In addition, we found the synergistic effect in combining the ST1814G/Fiber-2 yeast and inactivated vaccine to trigger stronger humoral immunity and mucosal immunity. Our results suggest that oral live ST1814G/Fiber-2 is a potentially safer auxiliary preparation strategy in controlling FAdV-4 infection.

Improving Control of Gene Therapy-Based Neurotrophin Delivery for Inner Ear Applications.

Background: Survival and integrity of the spiral ganglion is vital for hearing in background noise and for optimal functioning of cochlear implants. Numerous studies have demonstrated that supplementation of supraphysiologic levels of the neurotrophins BDNF and NT-3 by pumps or gene therapy strategies supports spiral ganglion survival. The endogenous physiological levels of growth factors within the inner ear, although difficult to determine, are likely extremely low within the normal inner ear. Thus, novel approaches for the long-term low-level delivery of neurotrophins may be advantageous. Objectives: This study aimed to evaluate the long-term effects of gene therapy-based low-level neurotrophin supplementation on spiral ganglion survival. Using an adenovirus serotype 28-derived adenovector delivery system, the herpes latency promoter, a weak, long expressing promoter system, has been used to deliver the BDNF or NTF3 genes to the inner ear after neomycin-induced ototoxic injury in mice. Results: Treatment of the adult mouse inner ear with neomycin resulted in acute and chronic changes in endogenous neurotrophic factor gene expression and led to a degeneration of spiral ganglion cells. Increased survival of spiral ganglion cells after adenoviral delivery of BDNF or NTF3 to the inner ear was observed. Expression of BDNF and NT-3 could be demonstrated in the damaged organ of Corti after gene delivery. Hearing loss due to overexpression of neurotrophins in the normal hearing ear was avoided when using this novel vector–promoter combination. Conclusion: Combining supporting cell-specific gene delivery via the adenovirus serotype 28 vector with a low-strength long expressing promoter potentially can provide long-term neurotrophin delivery to the damaged inner ear.

The Cellular and Viral circRNAs Induced by Fowl Adenovirus Serotype 4 Infection.

Circular RNAs (circRNAs) are a new class of noncoding RNAs that play vital roles in many biological processes. Virus infection induces modifications in cellular circRNA transcriptomes and expresses viral circRNAs. The outbreaks of Hydropericardium-hepatitis syndrome (HHS) caused by fowl adenovirus serotype 4 (FAdV-4) have resulted in huge economic losses to the poultry industry worldwide. To investigate the expression of circRNAs during FAdV-4 infection, we performed transcriptome analysis of FAdV-4-infected leghorn male hepatoma (LMH) cells. In total, 19,154 cellular circRNAs and 135 differentially expressed (DE) cellular circRNAs were identified. The characteristics of the DE cellular circRNAs were analyzed and most of them were related to multiple biological processes according to GO and KEGG enrichment analysis. The accuracy of 10 cellular circRNAs were verified by semiquantitative RT-PCR and sequencing. The change trend was consistent with the RNA sequencing results. Moreover, 2014 viral circRNAs were identified and 10 circRNAs were verified by the same methods. Our analysis showed that seven circRNAs with the same 3′ terminal and variable 5′ terminal regions were located at pTP protein and DNA pol protein of FAdV-4, which may be generated via alternative splicing events. Moreover, the expression level of viral circRNAs was closely related to the replication efficiency of the virus and partial of the viral circRNAs promoted the replication of FAdV-4. Competing endogenous RNA analysis further showed that the effects of cellular and viral circRNAs on host or viral genes may act via miRNAs. Collectively, our findings first indicate that FAdV-4 infection induced the differential expression of cellular circRNAs and FAdV-4 also expressed viral circRNAs, some of which affected FAdV-4 replication. These findings will provide new clues for further understanding FAdV-4 and provide a basis for investigating host-virus interactions.

Genetic diversity of fowl adenovirus serotype 8b isolated from cases of inclusion body hepatitis in commercial broiler chickens

Inclusion body hepatitis (IBH) is a devastating disease of chickens caused by Fowl adenovirus (FAdV) and molecular studies of hexon gene is important for classification of FAdV isolates, epidemiology of IBH infection and development of effective control strategy. The objective of the study was to molecularly characterize FAdV isolates obtained from the field outbreaks of IBH in broiler chicken from 2017 to 2019. Liver and gizzard samples named UPM1701, UPM1801, UPM1802, UPM1901 and UPM1902 were collected and processed for FAdV detection and characterization by PCR. Swollen, necrotic and haemorrhagic liver; erosion of koilin layer of gizzard and enlargement of the proventriculus were observed. The samples were positive for FAdV with 98% to 100% identical to serotype 8b reference strains based on NCBI Blast. Thirty-one nucleotide changes that produced 10 amino acid substitutions were observed in L1 loop region of UPM1701, UPM1901 and UPM1902 isolates. All isolates clustered together with FAdV 8b reference strains and shared common ancestor with UPM1137E2 and UPM04217. Gizzard erosion in FAdV 8b infection is not common and conciding with mutations indicate evolving novel pathogenicity. These mutations could have effect on the epidemiology of IBH and be useful in designing effective prevention and control strategy.

Hepatitis of Unknown Origin and Etiology (Acute Non HepA-E Hepatitis) among Children in 2021/2022: Review of the Current Findings

Several clusters and individual cases of acute hepatitis have been reported in the US, Europe and recently in Asia and Central America since October 2021. A laboratory investigation of the common viral hepatitis agents (HAV, HBV, HCV, HDV and HEV) yielded negative results prompting the use of the term “acute non HepA-E hepatitis” to describe this condition. The cases were characterized by the manifestations of acute hepatitis (abdominal pain, vomiting, diarrhea, jaundice and very high levels of liver enzymes) affecting children with a median age of 3–4 years. The exact underlying etiology has not been revealed yet; however, a leading hypothesis is that an infectious agent is the culprit, underlying cause or a risk factor for acute non HepA-E hepatitis occurrence. So far, laboratory testing has shown the presence of the group F human adenovirus serotype 41 (HAdV-F41) in about three-fourths of the investigated cases. As of 13 May 2022, more than 450 cases were reported worldwide, the majority of which were in the UK (n = 176), the US (n = 109), 13 European countries (at least 103 cases) and in Argentina, Brazil, Canada, Costa Rica, Indonesia, Israel, Japan, Palestine, Panama, Singapore and South Korea. Vigilant surveillance and epidemiologic investigations to identify further cases are warranted to delineate the features of this emergent public health issue. The possible role of environmental and toxic agents including foodborne toxins should also be considered. Specific guidelines for identification of further cases are necessary, particularly in low-income settings where testing for adenoviruses is not considered routinely. A genetic analysis of HAdV-F41 isolates is recommended to assess the potential changes in the virus genome with subsequent possible altered virus behavior. Immunopathogenesis is another possibility that should be evaluated considering the lack of viral structures in liver biopsies of the affected children in the US.

Arginine regulates inflammation response-induced by Fowl Adenovirus serotype 4 via JAK2/STAT3 pathway

BackgroundFowl Adenovirus serotype 4 (FAdV-4) infection causes severe inflammatory response leading to hepatitis-hydropericardium syndrome (HHS) in poultry. As an essential functional amino acid of poultry, arginine plays a critical role in anti-inflammatory and anti-oxidative stress.ResultsIn this study, the differential expression genes (DEGs) were screened by transcriptomic techniques, and the DEGs in gene networks of inflammatory response-induced by FAdV-4 in broiler’s liver were analyzed by Kyoto encyclopedia of genes and genomes (KEGG) enrichment. The results showed that the cytokines pathway and JAK/STAT pathway were significantly enriched, in which the DEGs levels of IL-6, IL-1β, IFN-α, JAK and STAT were significantly up-regulated after FAdV-4 infection. It was further verified with real-time fluorescence quantitative polymerase chain reaction (Real-time qPCR) and Western blotting (WB) in vitro and in vivo. The findings demonstrated that FAdV-4 induced inflammatory response and activated JAK2/STAT3 pathway. Furthermore, we investigated whether arginine could alleviate the liver inflammation induced by FAdV-4. After treatment with 1.92% arginine level diet to broilers or 300 μg/mL arginine culture medium to LMH cell line with FAdV-4 infection at the same time, we found that the mRNA levels of IL-6, IL-1β, IFN-α and the protein levels of p-JAK2, p-STAT3 were down-regulated, compared with FAdV-4 infection group. Furthermore, we confirmed that the inflammation induced by FAdV-4 was ameliorated by pre-treatment with JAK inhibitor AG490 in LMH cells, and it was further alleviated in LMH cells treatment with AG490 and ARG.ConclusionsThese above results provide new insight that arginine protects hepatocytes against inflammation induced by FAdV-4 through JAK2/STAT3 signaling pathway.

The Role of Hexon Amino Acid 188 Varies in Fowl Adenovirus Serotype 4 Strains with Different Virulence.

ABSTRACTHepatitis-hydropericardium syndrome (HHS) induced by fowl adenovirus serotype 4 (FAdV-4) has caused huge economic losses to poultry industries. The key genes responsible for different virulence of FAdV-4 strains are not fully elucidated. Previous studies indicated that hexon of pathogenic FAdV-4 has a conserved arginine (R) at position 188, and a conserved isoleucine (I) is present at this position in reported nonpathogenic FAdV-4. Recently, it was reported that R188 of hexon is the determinant site for pathogenicity of the emerging Chinese FAdV-4 strain. However, the role of hexon amino acid 188 (aa188) has not been examined in the nonpathogenic FAdV-4 strain. In this study, three recombinant FAdV-4 viruses, H/H/R188I, O/O/I188R, and H/O/I188R, were constructed by mutating hexon aa188 of FAdV-4 pathogenic strain CH/HNJZ/2015 (H) and nonpathogenic strain ON1 (O), and pathogenicity was assessed in specific-pathogen-free (SPF) chickens. Consistent with previous findings, H/O/I188R exhibited pathogenicity similar to that of CH/HNJZ/2015, yet H/H/R188I induced no mortality. Unexpectedly, all chickens infected with O/O/I188R survived. Postmortem examination of O/O/I188R-infected chickens showed typical lesions of inclusion body hepatitis rather than HHS. Expression of proinflammatory cytokines in CH/HNJZ/2015- and H/O/I188R-infected chickens was significantly higher than that in H/H/R188I-, ON1-, and O/O/I188R-infected chickens. Analysis of predicted hexon protein structures indicated that aa188 mutation leads to conformational changes in the L1 loop of HNJZ-hexon but not in ON1-hexon. In summary, the present study demonstrated that the role of hexon aa188 in the virulence of FAdV-4 varies between different strains. Induction of HHS requires factors aside from hexon aa188 in the emerging Chinese FAdV-4 strain.IMPORTANCE HHS induced by FAdV-4 has caused huge economic losses to the poultry industry. The key determinants for the different virulence of FAdV-4 have not been fully elucidated. Here, we investigated the role of hexon aa188 in FAdV-4 strains with different virulence and showed that the role of hexon aa188 varies in FAdV-4 strains with different genetic contents. The hexon R188 may be the key amino acid for causing inclusion body hepatitis by the pathogenic FAdV-4 strain, and induction of HHS by FAdV-4 may need other viral cofactors. Moreover, the hexon R188I mutation greatly affected the expression of proinflammatory cytokines induced by the pathogenic strain CH/HNJZ/2015, but no significant difference was observed between the nonpathogenic strain ON1 and ON1 with hexon I188R mutation. We found that hexon aa188 mutation induced conformational changes to hexon protein in CH/HNJZ/2015 but not in ON1, which might be the underlying reason for the changing virulence.

Therapeutic efficacy of an Ad26/MVA vaccine with SIV gp140 protein and vesatolimod in ART-suppressed rhesus macaques

Developing an intervention that results in virologic control following discontinuation of antiretroviral therapy (ART) is a major objective of HIV-1 cure research. In this study, we investigated the therapeutic efficacy of a vaccine consisting of adenovirus serotype 26 (Ad26) and modified vaccinia Ankara (MVA) with or without an SIV Envelope (Env) gp140 protein with alum adjuvant in combination with the TLR7 agonist vesatolimod (GS-9620) in 36 ART-suppressed, SIVmac251-infected rhesus macaques. Ad26/MVA therapeutic vaccination led to robust humoral and cellular immune responses, and the Env protein boost increased antibody responses. Following discontinuation of ART, virologic control was observed in 5/12 animals in each vaccine group, compared with 0/12 animals in the sham control group. These data demonstrate therapeutic efficacy of Ad26/MVA vaccination with vesatolimod but no clear additional benefit of adding an Env protein boost. SIV-specific cellular immune responses correlated with virologic control. Our findings show partial efficacy of therapeutic vaccination following ART discontinuation in SIV-infected rhesus macaques.

Burden, Clinical Characteristics, Risk Factors, and Seasonality of Adenovirus 40/41 Diarrhea in Children in Eight Low-Resource Settings

BackgroundThe application of molecular diagnostics has identified enteric group adenovirus serotypes 40 and 41 as important causes of diarrhea in children. However, many aspects of the epidemiology of adenovirus 40/41 diarrhea have not been described.MethodsWe used data from the 8-site Etiology, Risk Factors, and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development Project birth cohort study to describe site- and age-specific incidence, risk factors, clinical characteristics, and seasonality.ResultsThe incidence of adenovirus 40/41 diarrhea was substantially higher by quantitative polymerase chain reaction than enzyme immunoassay and peaked at ∼30 episodes per 100 child-years in children aged 7–15 months, with substantial variation in incidence between sites. A significant burden was also seen in children 0–6 months of age, higher than other viral etiologies with the exception of rotavirus. Children with adenovirus 40/41 diarrhea were more likely to have a fever than children with norovirus, sapovirus, and astrovirus (adjusted odds ratio [aOR], 1.62; 95% CI, 1.16–2.26) but less likely than children with rotavirus (aOR, 0.66; 95% CI, 0.49–0.91). Exclusive breastfeeding was strongly protective against adenovirus 40/41 diarrhea (hazard ratio, 0.64; 95% CI, 0.48–0.85), but no other risk factors were identified. The seasonality of adenovirus 40/41 diarrhea varied substantially between sites and did not have clear associations with seasonal variations in temperature or rainfall.ConclusionsThis study supports the situation of adenovirus 40/41 as a pathogen of substantial importance, especially in infants. Fever was a distinguishing characteristic in comparison to other nonrotavirus viral etiologies, and promotion of exclusive breastfeeding may reduce the high observed burden in the first 6 months of life.

Preparation, Characterization, and Antimicrobial and Antiviral Properties of Silver-Containing Nanocomposites Based on Polylactic Acid-Chitosan.

Antimicrobial and antiviral nanocomposites based on polylactic acid (PLA) and chitosan were synthesized by a thermochemical reduction method of Ag+ ions in the PLA-Ag+-chitosan polymer films. Features of the structural, morphological, thermophysical, antimicrobial, antiviral, and cytotoxic properties of PLA-Ag-chitosan nanocomposites were studied by X-ray diffraction (XRD), transmission electron microscopy (TEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and antiviral, antimicrobial, and cytotoxic studies. The effects of temperature and the duration of reduction of Ag+ ions on the structure of PLA-Ag-chitosan nanocomposites were established. During the thermochemical reduction (T = 160 °C, t = 5 min) of silver palmitate ions in PLA-Ag+-chitosan polymer films, Ag nanoparticles with an average size of 4.2 nm were formed. PLA-Ag-chitosan polymer nanocomposites have strong antimicrobial activity against S.aureus and E.coli strains. In particular, for PLA-chitosan samples containing 4% Ag, the diameters of the S.aureus and E.coli growth inhibition zones were 25.8 and 25.0 mm, respectively. The antiviral activity of the nanocomposites against influenza A virus, herpes simplex virus type 1, and adenovirus serotype 2 was also revealed. The PLA-4%Ag-chitosan nanocomposites completely inhibited the cytopathic effect (CPE) of herpes virus type 1 by 5.12 log10TCID50/mL (high antiviral activity) and the development of the CPE of influenza virus and adenovirus by 0.60 and 1.07 log10TCID50/mL (relative antiviral activity). The obtained nanocomposites were not cytotoxic; they did not inhibit the viability of MDCK, BHK-21, and Hep-2 cell cultures.

Gga-miR-30c-5p Enhances Apoptosis in Fowl Adenovirus Serotype 4-Infected Leghorn Male Hepatocellular Cells and Facilitates Viral Replication through Myeloid Cell Leukemia-1.

Fowl adenovirus serotype 4 (FAdV-4) is the primary causative agent responsible for the hepatitis-hydropericardium syndrome (HHS) in chickens, leading to considerable economic losses to stakeholders. Although the pathogenesis of FAdV-4 infection has gained attention, the underlying molecular mechanism is still unknown. Here, we showed that the ectopic expression of gga-miR-30c-5p in leghorn male hepatocellular (LMH) cells enhanced apoptosis in FAdV-4-infected LMH cells by directly targeting the myeloid cell leukemia-1 (Mcl-1), facilitating viral replication. On the contrary, the inhibition of endogenous gga-miR-30c-5p markedly suppressed apoptosis and viral replication in LMH cells. Importantly, the overexpression of Mcl-1 inhibited gga-miR-30c-5p or FAdV-4-induced apoptosis in LMH cells, reducing FAdV-4 replication, while the knockdown of Mcl-1 by RNAi enhanced apoptosis in LMH cells. Furthermore, transfection of LMH cells with gga-miR-30c-5p mimics enhanced FAdV-4-induced apoptosis associated with increased cytochrome c release and caspase-3 activation. Thus, gga-miR-30c-5p enhances FAdV-4-induced apoptosis by directly targeting Mcl-1, a cellular anti-apoptotic protein, facilitating FAdV-4 replication in host cells. These findings could help to unravel the mechanism of how a host responds against FAdV-4 infection at an RNA level.