Human Adenovirus Serotype Research Articles

Adenovirus-5 and adenovirus-37 seropositivity in obese patients

Background and Objectives: Obesity is a major health issue linked to conditions like diabetes, hypertension, and hy- perlipidemia. Infectobesity suggests that certain microorganisms may contribute to obesity. Human adenovirus serotypes, particularly Human adenovirus type-36 (HAdV-36), Human adenovirus type-5 (HAdV-5), and Human adenovirus type-37 (HAdV-37), are thought to influence body fat regulation. This study investigates the relationship between Immunoglobulin G (IgG) positivity for HAdV-5 and HAdV-37 and obesity, aiming to provide data on the infectious etiology of obesity. Materials and Methods: Blood samples separated into serums from obese (BMI ≥30) and non-obese (BMI 18.5-25) indi- viduals were tested for HAdV-5 and HAdV-37 seropositivity using ELISA kits and seropositivity rates between the groups were compared. Results: HAdV-37 antibody positivity was significantly higher in obese patients (39/48) compared to the control group (24/42) (p=0.011). For HAdV-5, antibody positivity was similar in both groups (26 individuals each) with no significant difference (p=0.461). No significant gender-related differences were found for either serotype. Conclusion: The study suggests HAdV-37 may be associated with obesity, while no such relationship was found for HAdV- There was no gender association for either serotype. These results align with existing literature on HAdV-37, but further research is needed to confirm the link between adenoviruses and obesity and explore potential treatment options.

Intranasal delivery of a recombinant adenovirus vaccine encoding the PEDV COE elicits potent mucosal and systemic antibody responses in mice.

Porcine epidemic diarrhea virus (PEDV) is an enteropathogenic coronavirus that causes substantial economic loss to the global pig industry. The emergence of PEDV variants has increased the need for new vaccines, as commercial vaccines confer inferior protection against currently circulating strains. It is well established that the induction of mucosal immunity is crucial for PEDV vaccines to provide better protection against PEDV infection. In this study, we constructed a recombinant adenovirus expressing the core neutralization epitope (COE) of G2b PEDV based on human adenovirus serotype 5 (Ad5). We evaluated the effects of different administration routes and doses of vaccine immunogenicity in Balb/c mice. Both intramuscular (IM) and intranasal (IN) administration elicited significant humoral responses, including COE-specific IgG in serum and mucosal secretions, along with serum-neutralizing antibodies. Moreover, IN delivery was more potent than IM in stimulating IgA in serum and mucosal samples and in dampening the immune response to the Ad5 vector. The immune response was stronger after high versus low dose IM injection, whereas no significant difference was observed between high and low IN doses. In summary, our findings provide important insights for developing novel PEDV vaccines.IMPORTANCEPorcine epidemic diarrhea (PED) is a highly contagious disease that has severe economic implications for the pork industry. Developing an effective vaccine against PEDV remains a necessity. Here, we generated a recombinant adenovirus vaccine based on Ad5 to express the COE protein of PEDV (rAd5-PEDV-COE) and systematically evaluated the immunogenicity of the adenovirus-vectored vaccine using different administration routes (intramuscular and intranasal) and doses in a mouse model. Our results show that rAd5-PEDV-COE induced potent systemic humoral response regardless of the dose or immunization route. Notably, intranasal delivery was superior to induce peripheral and mucosal IgA antibodies compared with intramuscular injection. Our data provide valuable insights into designing novel PEDV vaccines.

Intranasal adenovirus-vectored Omicron vaccine induced nasal immunoglobulin A has superior neutralizing potency than serum antibodies

The upper respiratory tract is the initial site of SARS-CoV-2 infection. Nasal spike-specific secretory immunoglobulin A (sIgA) correlates with protection against Omicron breakthrough infection. We report that intranasal vaccination using human adenovirus serotype 5 (Ad5) vectored Omicron spike in people who previously vaccinated with ancestral vaccine could induce robust neutralizing sIgA in the nasal passage. Nasal sIgA was predominantly present in dimeric and multimeric forms and accounted for nearly 40% of total proteins in nasal mucosal lining fluids (NMLFs). A low-level IgG could also be detected in NMLFs but not IgM, IgD, and IgE. After a complete nasal wash, sIgA in the nasal passage could be replenished rapidly within a few hours. A comparison of purified paired serum IgA, serum IgG, and nasal sIgA from the same individuals showed that sIgA was up to 3-logs more potent than serum antibodies in binding to spikes and in neutralizing Omicron subvariants. Serum IgG and IgA failed to neutralize XBB and BA.2.86, while nasal sIgA retained potent neutralization against these newly emerged variants. Further analysis showed that sIgA was more effective than IgG or IgA in blocking spike-mediated cell-to-cell transmission and protecting hACE2 mice from XBB challenge. Using a sIgA monoclonal antibody as a reference, we estimated that the total nasal sIgA contains about 2.6–3.9% spike-specific sIgA in NMLFs collected approximately one month after intranasal vaccination. Our study provided insights for developing intranasal vaccines that can induce sIgA to build an effective and mutation-resistant first-line immune barrier against constantly emerging variants.

Development of a novel adenovirus serotype 35 vector vaccine possessing an RGD peptide in the fiber knob and the E4 orf 4, 6, and 6/7 regions of adenovirus serotype 5

Adenovirus (Ad) vectors based on human adenovirus serotype 5 (Ad5) have attracted significant attention as vaccine vectors for infectious diseases. However, the effectiveness of Ad5 vectors as vaccines is often inhibited by the anti-Ad5 neutralizing antibodies retained by many adults. To overcome this drawback, we focused on human adenovirus serotype 35 (Ad35) vectors with low seroprevalence in adults. Although Ad35 vectors can circumvent anti-Ad5 neutralizing antibodies, vector yields of Ad35 vectors are often inferior to those of Ad5 vectors. In this study, we developed novel Ad35 vectors containing the Ad5 E4 orf 4, 6, and 6/7 or the Ad5 E4 orf 6 and 6/7 for efficient vector production, and compared their properties. These E4-modified Ad35 vectors efficiently propagated to a similar extent at virus titers comparable to those of Ad5 vectors. An Ad35 vector containing the Ad5 E4 orf 4, 6, and 6/7 mediated more efficient transduction than that containing the Ad5 E4 orf 6 and 6/7 in human cultured cells. Furthermore, insertion of an arginine-glycine-aspartate (RGD) peptide in the fiber region of an Ad35 vector containing the Ad5 E4 orf 4, 6, and 6/7 significantly improved the transgene product-specific antibody production following intramuscular administration in mice. The Ad35 vector containing the RGD peptide mediated efficient vaccine effects even in the mice pre-immunized with an Ad5.

Human Adenovirus Serotype 5 (HAdV-5) Infection of Kidney Fibroblast Cells Disrupts Cysteine, Purine, and Unsaturated Fatty Acid Metabolism in a Dose- and Time-Dependent Manner

Kidneys partake in numerous metabolic responsibilities, notably clearance of insulin from circulation, gluconeogenesis, and reabsorption of amino acids. Viral-induced kidney damage, either acutely or chronically, can occur from direct impact on kidney cells and from systemic and local immunological responses to counter the viral infection. Thus, it is likely that metabolic dysfunction is a consequence of infections. The detriments of viral infection on kidney cell metabolism remains an unelucidated area. We used human adenovirus serotype 5 (HAdV-5) to infect human embryonic kidney cells (HEK293T) in a dose- and time-dependent manner. We hypothesized that optimal metabolic dysfunction results after 24 hours of infection and at the highest viral dose. Cells were exposed to three viral dosages (0.5,1.0,2.0 multiplicity of infection [MOI]) and metabolism assessed by untargeted metabolomics at four time points (6-,12-,24-, 36-hours post infection [HPI]). A total of 182 metabolites were identified in our study. As early as 6 hrs, the metabolic profile of infected cells exhibited downregulation of cystathionine, cysteine, homocysteine, hypoxanthine, linoleic acid, and oleic acid with upregulation of 5'-Deoxy-5'-methylthioadenosine (MTA) and adenine. Peak dysregulation for carbohydrate, cysteine, organic acid, purine, and unsaturated fatty acid-related metabolites was observed at 12 hrs. At 24 hours, tapering of the overall metabolite response was observed with further dysregulation seen at 36 hours. Across all three dosages, responses to infection were similar. Perturbation of the cysteine, purine, and unsaturated fatty acid metabolic pathways has not previously been considered with HAdV-5 infection and a commonly used cell line exhibiting properties of kidney, and historically neuronal components. Although not investigated, uncovering adenoviral impact on enzymatic activity in these pathways could elucidate more understanding of renal impairment. These data altogether suggest that substantial DNA viral infection disrupts cellular metabolic pathways that may impair renal functions but need to be evaluated in an in vivo model. USDA HSI Educational Grant 2021-03397. Bailey-J Sanchez was supported by USDA HSI Educational Grant 2021-03397. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Comparative toxicological-hygienic assessment, structural-morphological, physicochemical characteristics, and virucidal properties of new nanopowder materials TiO2 and TiO2@Ag

In order to address safety concerns related to the acquisition and utilization of TiO2 and TiO2@Ag nanomaterials, as well as to investigate their disinfectant and biological effects, the structural-morphological, morpho­metry, toxicological, cytotoxic, and virucidal properties of these specified nanomaterials have been studied through experiments conducted on laboratory animals and in vitro. It has been demonstrated that the TiO2@Ag nanocomposite exhibited distinct physicochemical characteristics: it consisted of TiO2 nanoparticles ranging in size from 13 nm to 20 nm and Ag nanoparticles ranging from 35 nm to 40 nm with 4.0 wt% of silver localized on the surface of titanium dioxide. The purity of the modification of synthesized nano-TiO2 and nano-TiO2@Ag has been confirmed. Acute intraperitoneal administration of nanopowders revealed LD50 values of 4783.30 mg/kg for nano-TiO2 and 724.44 mg/kg for nano-TiO2@Ag. A slight accumulation was observed upon repeated (28-fold) intragastric administration of nano-TiO2. The cumulative dose administered, which equated to 15.9 multiples of the LD50 (76040 mg/kg), did not result in animal mortality but led to retardation in body weight gain. TiO2 and TiO2@Ag nanopowders do not irritate the skin, induce mild conjunctival irritation, and may exhibit a weak sensitizing effect. Nano-TiO2 and nano-TiO2@Ag powders accumulate in the tissues of internal organs and cause damage to the liver, kidneys, and lungs of laboratory animals upon intraperitoneal administration. The most characteristic morphological signs of the toxic effect of nano-TiO2 on liver tissue were observed at a level of 67.7% (cytoplasmic vacuolization in hepatocytes), while in the case of nano-TiO2@Ag initial necrotic changes were at a level of 70.0% (hepatocytes with pyknotic nuclei). Immunoassay analysis has demonstrated that TiO2@Ag and TiO2 nanomaterials at concentrations of 30 µg/ml can enhance the functional activity of peripheral blood mononuclear cells in vitro by increasing the production of cytokines IL-1, IL-6, TNF-α, and IL-4 in donors (p<0.05). This indicates the potential for chronic inflammation and allergic reactions among synthesis operators. In the study of the impact of nanomaterials on murine germ cells, it has been established that they affect the activity of mitochondrial enzymes and exert a damaging effect on mitochondrial membranes and overall cell integrity. Estimated approximate safe exposure levels in the workplace air are 0.3 mg/m3 for nano-TiO2 and 0.2 mg/m3 for nano-TiO2@Ag. Nano-TiO2@Ag and nano-TiO2 at a concentration of 100 µg/ml exhibit pronounced extracellular virucidal activity against human adenovirus serotype 2. The TiO2@Ag nanocomposite has a less damaging effect on Нер-2 cells compared to nano-TiO2.

Abstract 7241: Combination of CDK4/6 inhibitor promotes the antitumor effect of oncolytic adenovirus against canine breast cancer cells

Abstract Background: Oncolytic virotherapy has emerged as a novel antitumor platform to induce tumor-specific cytopathic effect against human cancer cells. However, whether oncolytic virotherapy is effective to canine cancer cells remains unclear. We developed telomerase-specific oncolytic adenoviruses OBP-301 and a p53-armed OBP-702 based on human adenovirus serotype 5. OBP-301 and OBP-702 have therapeutic potential against various types of human cancer cells. As recent report has suggested that human adenovirus serotype 5 induces cytopathic effect against canine cancer cells, OBP-301 and OBP-702 are expected to have therapeutic effects against canine cancer cells. Furthermore, it has been reported that CDK4/6 inhibitors clinically used for human breast cancers promote the replication of oncolytic adenoviruses, suggesting that combination of CDK4/6 inhibitors promote the therapeutic potential of oncolytic adenoviruses. In this study, we investigated the therapeutic potential of OBP-301 and OBP-702 against canine breast cancer cells in monotherapy and combination therapy with CDK4/6 inhibitor. Methods: Two canine breast cancer cell lines (CMT-U27, AZA-CB) were used in this study. The antitumor effect of OBP-301 and OBP-702 against canine breast cancer cells was analyzed using the XTT assay. The therapeutic potential of combination therapy with CDK4/6 inhibitor (palibociclib) was further analyzed. Virus-mediated induction of adenovirus E1A and p53 expression was analyzed by Western blotting. The in vivo antitumor effects of OBP-301 and OBP-702 were assessed using CMT-U27 subcutaneous tumor model. Results: OBP-301 and OBP-702 had cytopathic effect against CMT-U27 and AZA-CB cells. The antitumor activity of OBP-702 was superior to that of OBP-301. The combination of palbociclib and OBP-301 showed a more profound antitumor effect compared with monotherapy against CMT-U27 and AZA-CB cells. The expression of E1A and p53 was increased in virus-treated CMT-U27 and AZA-CB cells in a dose-dependent manner. Intratumoral administration of OBP-301 and OBP-702 significantly suppressed the growth of subcutaneous CMT-U27 tumors compared with mock treatment. Conclusion: Our results suggest that telomerase-specific oncolytic adenoviruses have cytopathic effect against canine breast cancer cells via virus infection and replication. Furthermore, combination of CDK4/6 inhibitor may be a promising strategy for promoting the antitumor effect of oncolytic adenovirus against canine breast cancers. Citation Format: Naoyuki Hashimoto, Hiroshi Tazawa, Satoru Kikuchi, Shinji Kuroda, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara. Combination of CDK4/6 inhibitor promotes the antitumor effect of oncolytic adenovirus against canine breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7241.

Respiratory adenovirus infections in children: a focus on Africa.

Lower respiratory tract infections (LRTIs) are an important cause of child morbidity and mortality globally, especially in children under the age of 5 years in Africa. Respiratory viruses, including human adenoviruses (HAdVs), are common causes of LRTIs in children. This review aims to shed light on the epidemiology, clinical manifestations, sequelae, and treatment options specific to adenovirus respiratory infections in African children. Recent evidence has challenged the perception that adenovirus is a negligible cause of LRTIs. Studies show HAdV emerging as the third most common viral pathogen in fatal pneumonias among under-5 children in low-income and middle-income African countries, contributing to 5.5% of all pneumonia deaths and ranking second in hospital-associated viral pneumonia deaths. Predominant HAdV serotypes associated with disease differ by country and region, and have changed over time. Risk factors for increased disease severity and long-term respiratory sequelae in previously healthy African children with HAdV LRTIs are not well established. Although respiratory viruses, including HAdV, are recognized contributors to LRTIs, the prevalence and impact of adenovirus infections have been under-recognized and understated. Available data suggests that African children, particularly those under 5 years old, are at risk of severe sequelae from respiratory HAdV infections. Long-term sequelae, including bronchiectasis and postinfectious bronchiolitis obliterans, further underscore the significant impact of HAdV infections. However, the scarcity of comprehensive data hampers our understanding of the extent of the impact of HAdV infections on child lung health in Africa. We recommend scaled-up HAdV surveillance, ensuring its consistent inclusion in population-level LRTI assessments, and expanded and equitable access to diagnostics for early recognition of African children at risk of developing chronic sequelae and death. Enhanced understanding of adenovirus epidemiology and clinical outcomes and the availability of therapeutic options are essential for informed public health strategies and clinical care.

Development of monoclonal antibodies targeting the conserved fragment of hexon protein to detect different serotypes of human adenovirus.

In this study, we selected the three most conserved antigenic fragments of human adenovirus to prepare a murine monoclonal antibody for the first time, and human adenovirus antigenic fragments with heretofore unheard of degrees of conservatism were isolated. The three monoclonal antibodies with the ability to recognize human respiratory adenovirus over a broad spectrum were screened by hybridoma and monoclonal antibody preparation. Human adenovirus infections are serious; however, therapeutic drugs and diagnostic reagents are scarce. Thus, to reduce the serious consequences of human viral infections and adenovirus pneumonitis, early diagnosis of infection is required. The present study provides three monoclonal antibodies capable of recognizing a wide range of human adenoviruses, thereby offering guidance for subsequent research and development.

High-Throughput Screening for the Prevalence of Neutralizing Antibodies against Human Adenovirus Serotype 5.

Adenoviral vectors based on the human adenovirus species C serotype 5 (HAdV-C5) are commonly used for vector-based gene therapies and vaccines. In the preclinical stages of development, their safety and efficacy are often validated in suitable animal models. However, pre-existing neutralizing antibodies may severely influence study outcomes. Here, we generated a new HAdV-C5-based reporter vector and established a high-throughput screening assay for the multivalent detection of HAdV-C5-neutralizing antibodies in serum. We screened the sera of rhesus macaques at different primate centers, and of rabbits, horses, cats, and dogs, showing that HAdV-C5-neutralizing antibodies can be found in all species, albeit at different frequencies. Our results emphasize the need to prescreen model animals in HAdV-C5-based studies.

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.

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).

Development of a Nanoplate-Based Digital PCR Test Method for Quantitative Detection of Human Adenovirus DNA.

Digital polymerase chain reaction (dPCR) assay is an advanced PCR technique that allows for the simultaneous detection and absolute quantification of diverse pathogens.Commercially validated kits available for detecting all subtypes of human adenovirus (HAdV) are limited. This study aimed to demonstrate the development of an in-house nanoplate-based dPCR assay with high sensitivity, even at low copy numbers. In this methodological study, the standardized HAdV DNA was prepared by amplifying the specific hexon gene region with real-time PCR and purifying the HAdV DNA using magnetic beads from HAdV-positive extractions. Dilutions were tested in triplicate during three independent runs to determine the dynamic range, the limit of detection (LoD), the limit of quantification (LoQ), precision, and reproducibility. The primer and probe sequences used in the study were selected based on a literature review to ensure the detection of all HAdV serotypes in a single run. The selected primers were verified using the US National Center for Biotechnology Information (NBCI) nBLAST tools, and the target sequence was determined using the BioEdit software. The DNA concentration of the stock solution was measured using a Qubit fluorometer. The estimated copy number of the stock solution per milliliter was calculated based on the length of the amplified base sequence and fluorometer measurement. The dynamic range of the test was determined to be from 770.4 to 0.9476 cp/μl, with the LoD and LoQ values both being 0.9476 cp/μl. The coefficient of determination (r 2) value of the test was 0.9986. The results demonstrated that the dPCR method could be an ideal tool for the diagnosis and absolute quantification of human adenoviruses, especially in low copy numbers. In order to determine the reproducibility of the test and validate the method for field use, it needs to be developed and adapted in various laboratories and supported by clinical studies.

Alterations in Lung Carcinoma Cells and Cell-Derived Extracellular Vesicles after Human Adenovirus Type 5 Administration

Human adenoviruses (HAdVs) affect the respiratory system of healthy individuals and people with pre-existing health conditions. These highly infectious viruses can alter a myriad of cellular and pathophysiological processes. HAdVs can modulate their infection ability and pathogenicity by regulating extracellular vesicles (EVs) formation and function. EVs are nanovesicles that facilitate intracellular communication, cell signaling/trafficking, and immune regulation. We propose that HAdVs can exploit EV formation, secretion, and release various pathways to promote infection and transmission between neighboring cells. In the present study, we explored the impact of HAdV serotype 5 (HAdV-5) on the A549-epithelial carcinoma cells and EVs derived from A549 cells. A549 cells were cultured in an exosome-depleted medium and administered different concentrations of HAdV-5 at varying time points. The effect of HAdV-5 infection on A549 cells and A549-derived EVs was determined by 4’,6-diamidino2-phenylindole (DAPI) stain, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay, electron microscopy, particle size, zeta potential analysis, immunosorbent analysis, total RNA levels, and total protein content. Our findings illustrated that HAdV-5 reduces A549-cell viability and promotes variations within the protein content of A549 cells. Our data explicitly demonstrated that HAdV-5 significantly altered EV numbers, biogenesis, and composition. These findings suggest that HAdV-5 modulates EVs’ biological properties and physiological functions in human lung carcinoma cells.

Silver and gold nanoparticles: Eco-friendly synthesis, antibiofilm, antiviral, and anticancer bioactivities

For the first time in this study, silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) were green synthesized by the cost-effective and eco-friendly procedure using Cotton seed meal and Fodder yeast extracts. The biosynthesized NPs were characterized by UV–Vis spectroscopy, dynamic light scattering analysis (DLS), transmission electron microscopy (TEM), selected area electron diffraction (SAED), and fourier-transform infrared (FTIR) spectroscopy. Furthermore, the biosynthesized NPs were tested in vitro against biofilm formation by some pathogenic negative bacteria (Escherichia coli, Proteus mirabilis, Klebsiella sp., Salmonella sp., and Pseudomonas aeruginosa) and negative bacteria (staphylococcus aureus) as well as against human denovirus serotype 5 (HAdV-5) and anticancer activity using HepG2 hepatocarcinoma cells. UV–Vis absorption spectra of reaction mixture of AgNPs and AuNPs exhibited maximum absorbance at 440 nm and 540 nm, respectively. This finding was confirmed by DLS measurements that the highest intensity of the AgNPs and AuNPs were 84 nm and 73.9 nm, respectively. FTIR measurements identified some functional groups detected in Cotton seed meal and Fodder yeast extracts that could be responsible for reduction of silver and gold ions to metallic silver and gold. The morphologies and particle size of AgNPs and AuNPs were confirmed by the TEM and SAED pattern analysis. Biosynthesized AgNPs and AuNPs showed good inhibitory effects against biofilms produced by Escherichia coli, Proteus mirabilis, Klebsiella sp., Salmonella sp., Pseudomonas aeruginosa, and Staphylococcus aureus. In addition, they showed anticancer activities against hepatocellular carcinoma (HepG-2) and antiviral activity against human adenovirus serotype 5 infection in vitro. Finally, the results of this study is expected to be extremely helpful to nano-biotechnology, pharmaceutical, and food packing applications through developing antimicrobial and/or an anticancer drugs from ecofriendly and inexpensive nanoparticles with multi-potentiality.

Development of oncolytic and gene therapy vectors based on adenovirus serotype 4 as an alternative to adenovirus serotype 5.

Adenoviral vectors are among the most frequently used vectors for gene therapy and cancer treatment. Most vectors are derived from human adenovirus (Ad) serotype 5 despite limited applicability caused by pre-existing immunity and unfavorable liver tropism, whereas the other more than 100 known human serotypes remain largely unused. Here, we screened a library of human Ad types and identified Ad4 as a promising candidate vector. Reporter-gene-expressing viruses representative of the natural human Ad diversity were used to transduce an array of muscle cell lines and two- or three-dimensional tumor cultures. The time-course of transgene expression was monitored by fluorescence or luminescence measurements. To generate replication-deficient Ad4 vector genomes, successive homologous recombination was applied. Ad4, 17 and 50 transduced human cardiomyocytes more efficiently than Ad5, whereas Ad37 was found to be superior in rhabdomyocytes. Despite its moderate transduction efficiency, Ad4 showed efficient and long-lasting gene expression in papillomavirus (HPV) positive tumor organoids. Therefore, we aimed to harness the potential of Ad4 for improved muscle transduction or oncolytic virotherapy of HPV-positive tumors. We deleted the E1 and E3 transcription units to produce first generation Ad vectors for gene therapy. The E1- and E1/E3-deleted vectors were replication-competent in HEK293 cells stably expressing E1 but not in the other cell lines tested. Furthermore, we show that the Ad5 E1 transcription unit can complement the replication of E1-deleted Ad4 vectors. Our Ad4-based gene therapy vector platform contributes to the development of improved Ad vectors based on non-canonical serotypes for a broad range of applications.

Genetic Evolution and Variation of Human Adenovirus Serotype 31 Epidemic Strains in Beijing, China, during 2010-2022.

Human adenovirus serotype 31 (HAdV-31) is closely associated with gastroenteritis in children and can cause fatal systemic disseminated diseases in immunocompromised patients. The lack of genomic data for HAdV-31, especially in China, will greatly limit research on its prevention and control. Sequencing and bioinformatics analyses were performed for HAdV-31 strains from diarrheal children in Beijing, China, during 2010-2022. Three capsid protein genes (hexon, penton, and fiber) were obtained in 37 cases, including one in which the whole genome was sequenced. HAdV-31 strains clustered into three distinct clades (I-III) in a phylogenetic tree constructed based on concatenated genes and the whole genome; the endemic strains only gathered into clade II, and most of the reference strains clustered into clade I. Compared with penton and hexon, fiber had a faster evolutionary rate (1.32 × 10-4 substitutions/site/year), an earlier divergence time (1697), lower homology (98.32-100% at the amino acid level), and greater genetic variation (0.0032). Four out of the six predicted positive selection pressure codons were also in the knob of fiber. These results reveal the molecular evolution characteristics and variations of HAdV-31 in Beijing, and fiber may be one of the main evolution driving forces.

Increased dose and duration of infection with human adenovirus serotype 5 shifts amino acid, nucleic acid, TCA, and carbohydrate metabolism in human kidney cells

Viruses are obligate intracellular parasites that overtake host cellular processes and establish an environment optimal for replication. Human adenoviruses are double stranded DNA viruses known to possess a unique tissue tropism including respiratory, gastrointestinal, ocular, and renal tissues. Furthermore, infection can result in mild to moderate symptoms with increased severity in those with dampened immunity. Disruptions to host metabolism is a well-known consequence of viral infection. Nonetheless, the relationship between adenoviruses and host metabolism remains a largely unelucidated area. Few studies have coupled metabolomic techniques with the examination of infection across the adenoviruses’ range of tissue targets. Fewer studies have investigated metabolic shifts with manipulation of viral dosage and duration of infection simultaneously. The objective for this study was to determine metabolic shifts in human kidney cells with different dosages of human adenovirus serotype 5 and couple this to duration of infection. It was hypothesized that prolonged viral infection with increased viral dosage would result in greater shifts in metabolites involved in amino acid metabolism, carbohydrate metabolism, nucleotide metabolism, and the tricarboxylic acid cycle. Three dosages of human adenovirus serotype 5 were used: 0.5. 1.0, and 2.0 multiplicity of infection. Infections were prolonged for 6, 12, 24, and 36 hours and cytosolic fractions were subsequently collected. Samples were then subjected to gas chromatography coupled to time-of-flight mass spectrometry. We found distinct separations in many primary carbon classes between uninfected controls and infected cells. Greater separation was seen in the higher dosed samples compared to the 0.5 MOI group. Analyses revealed 186 identified metabolites with significant upregulated and downregulated metabolites involved in nucleotide biosynthesis, amino acid metabolism, carbohydrate formation, and tricarboxylic acid cycle. Metabolites such as urea and pyrophosphate displayed the greatest degree of upregulation and downregulation, respectively. This was evident across all concentrations at 24 HPI. An increase in duration of infection was correlated with the emergence of more metabolites, with greater detection at increased viral concentration. Our results identify specific metabolic pathways perturbed by viral-host interactions, which have not been previously implicated in adenoviral infection and in kidney cells. Our study also provides a framework to investigate individual adenoviral protein contribution to metabolic disruptions. UC Merced USDA Hispanic Education Training Program Grant This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Modified adenovirus induces antigen specific T cells and effector memory T cells in mice

Abstract We engineered a novel adenovirus with a modified capsid for use as an integrated anticancer gene therapy and to circumvent pre-existing adenovirus immunity. The modified adenovirus, SynBAd, when administered intravenously, induced antigen-specific CD8 T cell immunity in the liver of mice (C57BL/6) and a splenic population of effector memory T cells at levels equivalent with those induced by human Adenovirus serotype 5 (Ad5). Although the virus elicited equivalent liver and splenic T cell responses, SynBAd did not elevate liver enzymes as commonly seen with Ad5. SynBAd bound factor X with similar affinity to Ad5, though it localized preferentially to the spleen relative to Ad5, suggesting greater systemic immune stimulation. CD8 T cells in the liver and spleen increased with SynBAd administration, indicating that the virus acts as a potent mobilizer of T cells. Although SynBAd is a potent immunogen, antibodies generated in mice immunized with Ad5 only weakly neutralized SynBAd infection in vitro, suggesting that preexisting Ad5 antibodies will not dramatically influence SynBAd efficacy. Localization of the adenovirus after administration, activation of antigen specific T cells, and mobilization of effector memory T cells, suggest that this approach will promote an antitumor immune response that could be used in multiple tumor indications.

In English

Zinc oxide nanostructures (NS) were grown on thin discontinuous films of noble metals of silver and gold in order to study their structure, optical properties as well as photocatalytic and antiviral activity. The paper presents the results of X-ray diffraction study, scanning electron microscopy study, photoluminescence and Raman measurements. X-ray diffraction experiments demonstrate similar patterns for all grown ZnO nanostructures. The SEM images of ZnO NS grown on Ag/Si and Au/Si wafers demonstrate more dense surface microstructure compared to ZnO NS grown on bare Si wafers. The most intensive ultraviolet and deep level emissions are observed for ZnO NS grown on Ag/Si wafers. Increase in thicknesses of Ag island film from 5 nm to 10 nm gives significant increase in intensity of ultraviolet and deep level emissions. Photocatalysis of grown ZnO nanostructures was studied by methyl orange dye degradation. Superior photocatalytic results are demonstrated by ZnO nanostructures grown on Ag/Si wafers, for which constants of dye degradation were twice higher than for ZnO nanostructures grown on Si and Au/Si substrates. The photocatalytic results correlates with photoluminescence spectra: more intensive photoluminescence in ultraviolet and visible ranges of optical spectrum leads to better photocatalytic performance. The cytotoxic effect of ZnO nanostructures was studied without photoactivation by the help of cell cultures MDCK and Hep-2 while the virucidal effect of ZnO nanostructures was studied by the help of Influenza A virus (H1N1) (strain FM / 1/47) and human adenovirus serotype 2 (HAdV2). ZnO nanostructures in a 1:10 dilution were not toxic to Hep-2 and MDCK cells. Most of the tested ZnO nanostructures exhibited no virucidal activity against human adenovirus serotype 2 (HAdV2) and influenza A virus (H1N1) (strain FM / 1/47) in the absence of photoexcitation.