OBJECTIVE: To construct an antisense RNA recombinant adenovirus vector of the human PDE5A1 promoter gene.. METHODS: A cDNA fragment containing the human PDE5A1 promoter and the human PDE5A1-specific exon was determined according to the gene bank. The antisense RNA fragment was synthesized artificially and subcloned into the pENTR. Then, the sequence of pENTR fragment was detected, and the recombinant adenovirus vector pAd/CMV/V5/antisense-PDE5A1 was constructed by LR reaction with the Gateway expression system. The identified recombinant adenovirus plasmid was digested with Pac I and transformed into 293A cells to package recombinant adenovirus particles. The recombinant adenovirus particles were tested with PCR technique and purified after acquisition by CsCl density gradient ultracentrifugation.. RESULTS: The sequencing result showed a 145 bp fragment in pENTR, which was proved to be the domain of the antisense RNA fragment. PCR confirmed that the antisense RNA fragment was cloned into the ...

TY - JOUR. T1 - Combination gene therapy with adenoviral vector-mediated HSV-tk + GCV and IL-12 in an orthotopic mouse model for prostate cancer. AU - Nasu, Y.. AU - Bangma, C. H.. AU - Hull, G. W.. AU - Yang, G.. AU - Wang, J.. AU - Shimura, S.. AU - Mccurdy, M. A.. AU - Ebara, S.. AU - Lee, H. M.. AU - Timme, T. L.. AU - Thompson, T. C.. N1 - Funding Information: This work was supported by grants from CaP CURE and NIH (CA68814 and SPORE P50-58204). We are most grateful to Drs Frank L Graham and Jack Gauldie for providing AdmIL-12 virus stock.. PY - 2001. Y1 - 2001. N2 - We previously demonstrated significant therapeutic activities associated with adenoviral vector-mediated Herpes Simplex Virus/thymidine kinase (AdHSV-tk) with ganciclovir (GCV) in situ gene therapy in the RM-1 orthotopic mouse prostate cancer model and interleukin-12 (AdmIL-12) in situ gene therapy in the RM-9 orthotopic mouse prostate model for prostate cancer. In both protocols, local cytotoxicity and activities against ...

TY - JOUR. T1 - Retrograde transfer of replication deficient recombinant adenovirus vector in the central nervous system for tracing studies. AU - Kuo, Hui. AU - Ingram, Donald K.. AU - Crystal, Ronald. AU - Mastrangeli, Andrea. PY - 1995/12/24. Y1 - 1995/12/24. N2 - We assessed the application of a replication deficient recombinant adenovirus vector as a retrograde tracer in neural pathway studies. The adenovirus vector, Ad.RSVBgal, containing the intracellular marker gene, β-galactosidase, was injected directly into the laterodorsal striatum of rats. The retrograde transport of the vector from the injection site was clearly visible in the cerebral cortex, thalamic nucleus, and substantia nigra. No evidence for anterograde transport of the vector was found. When the vector was injected into the genu of the corpus callosum, little uptake of the vector by fibers was noted which suggested that uptake by fibers-of-passage should not be a problem in tracing studies. The present study demonstrates ...

Title: Transductional Targeting with Recombinant Adenovirus Vectors. VOLUME: 2 ISSUE: 3. Author(s):Valerie Legrand, Philippe Leissner, Arend Winter, Majid Mehtali and Monika Lusky. Affiliation:CAREXS.A., 11 rue Humann, 67000 Strasbourg, France. Keywords:Adenovirus Vectors, TROPISM, Fiber Protein, Monoclonal antibody. Abstract: Replication-deficient adenoviruses are considered as gene delivery vectors for the genetic treatment of a variety of diseases. The ability of such vectors to mediate efficient expression of therapeutic genes in a broad spectrum of dividing and non-dividing cell types constitutes an advantage over alternative gene transfer vectors. However, this broad tissue tropism may also turn disadvantageous when genes encoding potentially harmful proteins (e.g. cytokines, toxic proteins) are expressed in surrounding normal tissues. Therefore, specific restrictions of the viral tropism would represent a significant technological advance towards safer and more efficient gene delivery ...

OBJECTIVES To investigate the validity of catalase recombinant adenovirus on the treatment of oxidative cataract. METHODS The coding sequence of catalase was cloned and the catalase recombinant adenovirus was constructed. The expression time course of catalase gene in rat lens infected by recombinant adenovirus was determined by Western blotting. Cultured rat lens were randomly divided into 3 groups: the control group, the group treated by hydrogen peroxide and the group treated by hydrogen peroxide combined with catalase recombinant adenovirus. The transparence and apoptosis ratio of lens on the time points of 6, 12, 18, 24 hours were determined by image analysis and double colour flowcytometry. RESULTS The coding sequence of catalase was cloned and recombinant adenovirus was successfully constructed. The expression of catalase in cultured rat lens infected by recombinant adenovirus reached peak point on 9 hours post infection and maintained the level in the whole experiment period. The

Improvement of transduction and augmentation of cytotoxicity are crucial for adenoviruses (Ad)-mediated gene therapy for cancer. Down-regulated expression of type 5 Ad (Ad5) receptors on human tumors hampered Ad-mediated transduction. Furthermore, a role of the p53 pathways in cytotoxicity mediated by replication-competent Ad remained uncharacterized. We constructed replication-competent Ad5 of which the E1 region genes were activated by a transcriptional regulatory region of the midkine or the survivin gene, which is expressed preferentially in human tumors. We also prepared replication-competent Ad5 which were regulated by the same region but had a fiber-knob region derived from serotype 35 (AdF35). We examined the cytotoxicity of these Ad and a possible combinatory use of the replication-competent AdF35 and Ad5 expressing the wild-type p53 gene (Ad5/p53) in esophageal carcinoma cells. Expression levels of molecules involved in cell death, anti-tumor effects in vivo and production of viral progenies

Granulocyte macrophage-colony stimulating factor (GM-CSF) is a hematopoietic growth factor involved in differentiation, survival and activation of myeloid and non-myeloid cells with important implications for lung antibacterial immunity. Here we examined the effect of pulmonary adenoviral vector-mediated delivery of GM-CSF (AdGM-CSF) on anti-mycobacterial immunity in M. bovis BCG infected mice. Exposure of M. bovis BCG infected mice to AdGM-CSF either applied on 6h, or 6h and 7days post-infection substantially increased alveolar recruitment of iNOS and IL-12 expressing macrophages, and significantly increased accumulation of IFNγpos T cells and particularly regulatory T cells (Tregs). This was accompanied by significantly reduced mycobacterial loads in the lungs of mice. Importantly, diphtheria toxin-induced depletion of Tregs did not influence mycobacterial loads, but accentuated immunopathology in AdGM-CSF-exposed mice infected with M. bovis BCG. Together, the data demonstrate that AdGM-CSF ...

TY - JOUR. T1 - Postischemic gene transfer of midkine, a neurotrophic factor, protects against focal brain ischemia. AU - Takada, J.. AU - Ooboshi, H.. AU - Ago, T.. AU - Kitazono, T.. AU - Yao, H.. AU - Kadomatsu, K.. AU - Muramatsu, T.. AU - Ibayashi, S.. AU - Iida, M.. PY - 2005/3/1. Y1 - 2005/3/1. N2 - Gene therapy may be a promising approach for treatment of brain ischemia. In this study, we examined the effect of postischemic gene transfer of midkine, a heparin-binding neurotrophic factor, using a focal brain ischemia model with the photothrombotic occlusion method. At 90 min after induction of brain ischemia in spontaneously hypertensive rats, a replication-deficient recombinant adenovirus encoding mouse midkine (AdMK, n = 7) or a control vector encoding β-galactosidase (Adβgal, n = 7) was injected into the lateral ventricle ipsilateral to ischemia. At 2 days after ischemia, we determined infarct volume by 2,3,5-triphenyltetrazolium chloride staining. There were no significant ...

Control of the HIV pandemic can only be achieved with the development of a safe and effective preventive HIV vaccine. A vaccine that will prevent HIV infection will elicit a strong immune response from both CD4 and CD8 cells. Recombinant adenovirus serotype vectors have been shown to elicit just such a response. The purpose of this study is to determine the safety and immunogenicity of the recombinant adenovirus serotype 5 preventive HIV-1 vaccine.. This study will last 18 to 24 months. Participants will be randomly assigned to one of four arms that will receive vaccine or placebo administered via intramuscular injection. Participants in Arms 1, 2, and 3 will all receive 3 injections. Participants in Arm 4 will receive one injection. For most participants, there will be 10 study visits in this study; for participants in Arm 4, there will be only 7 visits. For Arms 1, 2, and 3, study visits will occur at baseline and on Days 0, 14, 28, 42, 56, 168, 182,196, and 365. Participants in Arms 1, 2, and ...

Adenoviral Vectors for Gene treatment, moment Edition offers special, entire assurance of the gene supply cars which are in line with the adenovirus thats rising as an immense instrument in gene remedy. those fascinating new healing brokers have nice capability for the therapy of illness, making gene remedy a fast-growing box for learn. This ebook provides themes starting from the fundamental biology of adenoviruses, during the building and purification of adenoviral vectors, state-of-the-art vectorology, and using adenoviral vectors in preclinical animal types, with ultimate attention of the regulatory concerns surrounding human medical gene remedy trials. This extensive scope of knowledge offers a superb evaluate of the sector, permitting the reader to achieve an entire figuring out of the advance and use of adenoviral vectors.. ...

Intraperitoneal (i.p.) recurrence of cisplatin-refractory and p53 mutant ovarian cancer is a major clinical problem, despite surgery and chemotherapy. dl1520 (ONYX-015) is an E1B-55 kDa gene-deleted adenovirus engineered selectively to replicate in and destroy cancer cells lacking functional p53. Ho …

Recombinant adenovirus vectors are promising, highly characterized platforms from which novel vaccines can be produced. Conventional Ad5[E1−] vaccines possess the ability to promote strong immunologic responses against their expressed transgenes, but the same properties also trigger host antiviral responses, which can lead to increased toxicity, limited persistence, and decreased efficacy in the face of preexisting anti-Ad5 immunity (7, 28, 29, 34, 46). Alternatively, multiply deleted Ad5[E1−,E2b−] vaccines have been found to overcome several of these obstacles. Head-to-head comparisons of traditional Ad5[E1−] vaccines and E2b gene-deleted, Ad5[E1−,E2b−] vaccines have shown that the latter produce virtually no hepatotoxicity after systemic administration (29), persist and express transgenes for longer durations than Ad5[E1−] vaccines (28, 34), and remain therapeutically efficacious in hosts harboring substantial preexisting anti-Ad5 immunity (5, 14-16, 30).. It has been largely ...

TY - JOUR. T1 - An effective gene-knockdown using multiple shRNA-expressing adenovirus vectors. AU - Motegi, Yukari. AU - Katayama, Kazufumi. AU - Sakurai, Fuminori. AU - Kato, Takuya. AU - Yamaguchi, Tomoko. AU - Matsui, Hayato. AU - Takahashi, Masahide. AU - Kawabata, Kenji. AU - Mizuguchi, Hiroyuki. PY - 2011/7/30. Y1 - 2011/7/30. N2 - Viral vectors expressing short hairpin RNA (shRNA) are attractive for efficient and tissue-specific RNA interference (RNAi) delivery. We and others previously reported that recombinant adenovirus (Ad) vector-mediated RNAi has great potential for a variety of applications in molecular biology studies and gene therapy. In the present study, we have developed an efficient Ad vector-mediated RNAi system, in which an Ad vector carries four shRNA-expression cassettes (Ad-multi-shRNA vector), a simple and effective strategy for enhancing the RNAi response per Ad vector particle. The data demonstrated that the Ad-multi-shRNA vectors showed an enhanced RNAi effect ...

Recombinant adenoviruses currently are used for a variety of purposes, including gene transfer in vitro, vaccination in vivo, and gene therapy (1-4). Several features of adenovirus biology have made such viruses the vectors of choice for certain of these applications. For example, adenoviruses transfer genes to a broad spectrum of cell types, and gene transfer is not dependent on active cell division. Additionally, high titers of viruses and high levels of transgene expression generally can be obtained.. Decades of study of adenovirus biology have resulted in a detailed picture of the viral life cycle and the functions of the majority of viral proteins (5, 6). The genome of the most commonly used human adenovirus (serotype 5) consists of a linear, 36-kb, double-stranded DNA molecule. Both strands are transcribed and nearly all transcripts are heavily spliced. Viral transcription units are conventionally referred to as early (E1, E2, E3, and E4) and late, depending on their temporal expression ...

The plasminogen activation (PA) system is involved in vascular remodelling. Modulating its activity in vascular cells might be a way to interfere in processes such as angiogenesis and restenosis. Adenoviral vectors have become a favourable tool for direct gene transfer into vascular cells. In the interest of using adenoviral vectors to modulate plasminogen activator activity and endothelial and smooth muscle cell migration, we studied the effects of endothelial and smooth muscle cell transduction in vitro and in the umbilical vein ex vivo with a replication-defective adenoviral vector containing the β-galactosidase gene (AdCMVLacZ). Segments of the umbilical vein were infected with AdCMVLacZ (109-1010 pfu/ml). After 48 h strong β-galactosidase expression could be observed in the vessel wall, which was restricted to the endothelial layer. Although some heterogeneity in the transduction throughout the vessel could be seen, β-galactosidase expression was detectable for 21 days in explant. ...

A recombinant nucleic acid used for the production of a defective adenovirus containing an inserted sequence coding for a cytokine under the control of a promoter in the genomic sequence of the recombinant adenovirus. This recombinant adenovirus is useful in the preparation of anti-tumoral drugs which can be directly injected into the tumor of the host.

A simplified Ebola vaccine that consists of a modified GP protein (which is well-tolerated by human cells even at high concentrations) in a replication-defective adenoviral vector protects macaques.

La terapia génica consiste en la manipulación y utilización de material genético para el tratamiento de patologías. No obstante, esta estrategia requiere el uso de vectores de terapia génica para transportar el material genético al tejido diana de forma eficiente. Los vectores de terapia génica más comunes son los basados en el adenovirus humano de serotipo 5 (Ad5), porque, respecto a otros serotipos, el Ad5 tiene como ventaja su bioseguridad y facilidad de producción. Sin embargo, para que una célula sea transducida por el Ad5 ha de expresar un receptor reconocible por el Ad5, principalmente la proteína CAR, al que se une a través de su proteína fiber. El fiber de otros serotipos de adenovirus se une a receptores diferentes, permitiendo así la transducción de tipos celulares alternativos a los transducidos por el Ad5. De esta manera, una de las estrategias utilizadas para obtener un vector Ad5 con el tropismo de un serotipo diferente, es la pseudotipación del adenovirus, que ...

TY - JOUR. T1 - An altered subunit configuration associated with the actively transcribed DNA of integrated adenovirus genes. AU - Flint, S. J.. AU - Weintraub, Harold M.. PY - 1977/11. Y1 - 1977/11. N2 - The sensitivity to deoxyribonuclease I (DNAase I) of integrated adenovirus genes that encode mRNA has been compared to the sensitivity of adjacent viral DNA sequences that are not expressed as mRNA in two lines of adenovirus type 5-transformed hamster cells. We determined the concentrations of integrated DNA sequences homologous to different regions of the viral genome before and after mild DNAase I digestion of intact nuclei by measuring the rate of reassociation of restriction endonuclease fragments of labeled adenovirus DNA in the presence of DNA isolated from untreated and digested transformed cell nuclei. The HT14A cell line contains 2.4 copies of the left-hand 35% of the adenovirus type 5 genome per diploid quantity of cell DNA. Integrated sequences that are preferentially sensitive to ...

Wild-type p53 is involved in several aspects of cell cycle control and suppression of transformation, inducing either apoptosis or G1 block in cell cycle progression. Using a recombinant adenovirus containing the wild-type p53 cDNA, the biological effects of the newly expressed wild-type p53 protein were examined in six human glioma cell lines. Three cell lines (U-251 MG, U-373 MG, and A-172) expressed endogenous mutant p53, and the other three (U-87 MG, EFC-2, and D54 MG) expressed wild-type p53. The restoration of normal p53-encoded protein in the mutant cell lines induced apoptosis as assessed by morphological studies using nuclear staining, electron microscopy, and flow cytometric assays. In wild-type p53 cell lines, however, the overexpression of wild-type p53 did not result in apoptosis but inhibited cellular proliferation rather drastically and modified the neoplastic phenotype. Differential effects suggest two pathways for glioma oncogenesis and a possible therapeutic strategy.. ...

Dendritic cells (DCs) are professional antigen presenting cells that are being considered as potential immunotherapeutic agents to promote host immune responses against tumor antigens. The use of such modified antigen-presenting cells for research or therapeutic have been limited by several factors, including maintaining DCs in a highly activated state, efficient transduction and expression, stable expression, identification of appropriate tumor-associated antigens, and absence of unintended functional changes or cytotoxicity. In this study, the feasibility of using CD34-DCs for tumor immunotherapy after transduction with a recombinant adenovirus containing HBsAg gene (AdVHBsAg), an HCC-associated antigen, was investigated. The gene transfer with recombinant adenovirus vectors (AdV) can obtained high levels of stable expression of HBsAg and its efficiency was increased in a multiplicity of infection (MOI)-dependent manner. Moreover, the AdVHBsAg infection had no appreciable effect on apoptosis ...

Chronic obstructive pulmonary disease (COPD) and lung cancer are major causes of death in the world. Exposure to tobacco smoke presents the main factor in the development of COPD. Epidemiological research has shown that COPD represents, regardless of smoking status, the biggest risk factor for developing lung cancer. The genetic basis of the association of these pathological entities is unknown. The regulation of the immune response plays an important role in maintaining homeostasis. Toll-like receptors (TLR) are important for initiating immunity by recognising the sequences associated with the pathogen and inducing signal pathways in the host as an antimicrobial response. Previous researches have shown that the genetic variant of TLR5, N592S gene, is associated with COPD and lung cancer. The aim of this research was to construct a replication-defective adenoviral vector by introducing a gene for TLR5wt and TLR5N592S into the adenovirus type 5 (Ad5) backbone that would allow the insertion of ...

Biological Sciences Shirley, Cat.No. AD00876Z Description Adenovirus with ORF of aminolevulinate dehydratase (ALAD) with C terminal Flag and His tag....

Dendritic cells (DCs) modified by some immunomodulatory genes can stimulate a strong antitumor immunity and improve the treatment of tumor cells on the condition that the sources of tumor-associated antigens (TAAs) are available. IL-6, a pleotropic cytokine, has been found to inhibit CD4+25+ regulatory T (Treg)-cell-mediated immune suppression and decrease activation-induced cell death (AICD) without interfering the process of T-cell activation. To enhance DC-based cancer vaccine, we engineered DCs to express transgene IL-6.. We constructed a fiber-modified recombinant adenovirus vector AdVIL-6 expressing IL-6, infected DCs with AdVIL-6, and then investigated the efficacy of antitumor immunity induced by vaccination with DCs engineered to express IL-6 transgene. We demonstrated that DCs infected with the recombinant adenovirus AdVIL-6 induced DC maturation by up-regulation of the expression of MHC class U (Iab), CD40, CD54 and CD80 expression. We also demonstrated that vaccination of OVA-pulsed ...

Many applications for human gene therapy would be facilitated by high levels and long duration of physiologic gene expression. Adenoviral vectors are frequently used for gene transfer because of their high cellular transduction efficiency in vitro and in vivo. Expression of viral proteins and the lo …

Described are vaccines comprising recombinant vectors, such as recombinant adenoviruses. The vectors comprise heterologous nucleic acids encoding at least two antigens from one or more tuberculosis-c

13:2; potential epub Adenovirus notions; tenure bhakti 1? On the epub Adenovirus Methods and of this interest, are above under form. 1282 An epub Adenovirus Methods and Protocols: Adenoviruses, Ad study read by Porten - Szubin 1987:187.

A quantum dot method for highly efficient labelling of single adenoviral particles is developed. The technique has no impact on viral fitness and allows the imaging and tracking of virus binding and internalisation events using a variety of techniques including imaging cytometry and confocal microscopy. The method is applied to characterise the tropism of different adenoviral vectors.. ...

Gene Therapy adenoviral vectors explained, information about the mechanism of Adenoviruses, Adeviral particle organisation and genome organisation of adenoviruses

As combining therapeutic agents with different action mechanisms may enhance efficacy, YSC-02, an oncolytic adenovirus with multi targets was loaded with five different genes, which were designed with the expectation that different action mechanisms would cooperate. In spite of concerns regarding many APIs, YSC-02 was constructed to be an adenovirus-based anti-cancer drug. By using our own established mouse model system suitable for efficient adenoviral infection and replication, immune activity as well as survival potential could be precisely estimated for anti-cancer drug efficacy. YSC-02 was designed to decrease tumor cell survival, metastasis and to increase tumor cell death, and immune activation. It is composed of five different target genes, including one fused form and two shRNAs, but each of these genes functions is closely linked to produce the maximal antitumor effect. YSC-02 is like an organic complex designed to be applied primarily to heterogeneous liver cancer and melanoma. The ...

This study was initiated with the original aim of assessing the consequences of the expression of the functional α2-subunit of the rat Na+-K+-ATPase in the neonatal rat cardiac myocytes lacking this isoform. It soon became evident, however, that only a truncated α2-subunit not likely to be functional was overexpressed in these cells. Because enzyme and receptor fragments may often act like inactive mutant variants and cause dominant negative inhibition (2, 10, 12, 23, 27, 36) we attempted to determine whether the expression of the α2-fragment impaired the function of endogenous Na+-K+-ATPase in the neonatal myocytes. Our findings clearly show that the ion transport function of Na+-K+-ATPase is indeed inhibited concomitant with the expression of the truncated α2-isoform and that this is accompanied by a significant reduction of the α1-protein content of the neonatal myocyte. Because the induced reduction of the α3-protein content is small, if any, and because it is established that α1 ...

The first region 4 open reading frame 3 protein FLNA (E4-ORF3; UniProt Identification type:entrez-protein attrs :text:P04489″ term_id :119084″ term_text :P04489″P04489) may be the most extremely conserved of most adenovirus-encoded gene items on the amino acidity level. supplementary/tertiary framework and the capability to type heterogeneous higher-order multimers in option. Importantly a non-functional E4-ORF3 mutant proteins L103A forms a well balanced dimer with WT supplementary structure content. As the L103A mutant is certainly Coumarin 30 not capable of PML reorganization this result shows that higher-order multimerization of E4-ORF3 could be necessary for the experience of the proteins. To get this hypothesis we demonstrate the fact that E4-ORF3 L103A mutant proteins works as a dominant-negative effector when coexpressed using the WT E4-ORF3 in mammalian cells. It prevents WT E4-ORF3-mediated PML monitor development presumably by binding towards the WT proteins and ...

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Much concern has focused on the direct toxic effects of adenoviruses, particularly as intravenous administration of the virus can induce acute liver injury, as shown in animal models. It is this effect which may have triggered the cascade of events leading to the death of the patient with OTC deficiency-in this case the recombinant virus was injected directly into the hepatic artery. Studies in mice have highlighted the dose limiting liver toxicity of intravenously administered virus, which in this model is mainly due to an acute inflammatory response involving the release of certain cytokines (interleukin 6, interleukin 8, tumour necrosis factor α) and the recruitment of immune effector cells into the liver.5-7These effects are manifest within the first few hours of adenovirus administration and do not require de novo virus gene expression. A recent study demonstrated that adenovirus induced chemokine gene expression within the liver occurs within one hour after infection and results in the ...

Description: Accurate measurement of adenovirus titer is critical for gene delivery. Traditional plaque-forming unit (PFU) assays are long and suffer from high inter-assay variability. The QuickTiter Adenovirus Titer Immunoassay Kit provide a quick, complete system to functionally titer virus infectivity. The assay recognizes all 41 serotypes of adenovirus, and can be used with any adenovirus system that can amplify in HEK 293 cells ...

Adenoviruses are ubiquitous. Weve all been infected with lots of them (gets to be a problem when making gene therapy vectors). The older you are, the more likely you have been exposed to the different kinds of adenovirus, to the point where if you are an adult reading this, you probably have antibodies to serotypes 1, 2, 5, and/or 6 (depending on where you live).. Thus older children are more likely to have been exposed to any adenovirus, including Ad-36. If your obese group of children is 63% kids aged 15-18, and your healthy weight group is only 17% 15-18, I am not shocked at all that antibodies to Ad-36 is correlated with obesity. I bet you could have picked any other adenovirus serotype and found the same correlation. But they didnt look.. William M. Briggs gets it. I dunno the dude. Statistician. But he gets it. *thumbs-up*. From a virologists perspective, there are putative, plausible mechanisms for how Ad-36 could actually play a role in obesity. Those reasonings require ongoing ...

When walking through the hospital you sense the enormity of the task at hand for the medical teams. Families are forced to sleep in corridors, on stairwells and in the grounds of the hospital. There simply are not enough members of staff to deal with the number of children in need of care and support. Although things are difficult, Adil says his mother is helping him to cope.. World Child Cancers appeal to Give the Gift of Growing Up aims to give children with cancer, just like Adil, hope for the future. His mother, Shusmita, says she used to want her son to study hard and earn a scholarship for university, but she now just hopes she can see her son survive his cancer and go back to his normal life. However, Adil has bigger plans;. ...

Detect adenovirus rescue even before cytopathic effect (CPE) is observed. Ensure high titer by harvesting amplified adenovirus at the optimal time

Detect adenovirus rescue even before cytopathic effect (CPE) is observed. Ensure high titer by harvesting amplified adenovirus at the optimal time

TY - JOUR. T1 - Fiber-modified adenovirus for central nervous system Parkinsons disease gene therapy. AU - Lewis, Travis B.. AU - Glasgow, Joel N.. AU - Harms, Ashley S.. AU - Standaert, David G.. AU - Curiel, David T.. N1 - Copyright: Copyright 2015 Elsevier B.V., All rights reserved.. PY - 2014/8/21. Y1 - 2014/8/21. N2 - Gene-based therapies for neurological diseases continue to develop briskly. As disease mechanisms are elucidated, flexible gene delivery platforms incorporating transcriptional regulatory elements, therapeutic genes and targeted delivery are required for the safety and efficacy of these approaches. Adenovirus serotype 5 (Ad5)-based vectors can carry large genetic payloads to provide this flexibility, but do not transduce neuronal cells efficiently. To address this, we have developed a tropism-modified Ad5 vector with neuron-selective targeting properties for evaluation in models of Parkinson disease therapy. A panel of tropism-modified Ad5 vectors was screened for enhanced ...

Oncolytic adenoviruses are promising anticancer agents due to their ability to self-amplify at the tumor mass. In order to design a potent and selective oncolytic adenovirus that maintains intact all the viral functions with minimal increase in genome size we inserted palindromic E2F-binding sites into the endogenous promoter. The insertion of these sites controlling E1A-Δ24 results in a low systemic toxicity profile in mice. Importantly the E2F-binding sites also increased the cytotoxicity and the systemic antitumor activity relative to wild-type adenovirus in all cancer models tested. Pevonedistat The low toxicity and the increased potency results in improved antitumor efficacy after systemic injection and increased survival of mice transporting tumors. Furthermore the constrained genome size of this backbone allows an efficient and potent expression of transgenes indicating that this computer virus holds promise for overcoming the limitations of oncolytic adenoviral therapy. Introduction ...

TY - JOUR. T1 - Markedly enhanced cytolysis by E1B-19kD-deleted oncolytic adenovirus in combination with cisplatin. AU - Yoon, A. Rum. AU - Kim, Joo Hang. AU - Lee, Young Sook. AU - Kim, Hoguen. AU - Yoo, Ji Young. AU - Sohn, Joo Hyuk. AU - Park, Byeong Woo. AU - Yun, Chae Ok. PY - 2006/4/1. Y1 - 2006/4/1. N2 - Oncolytic adenoviruses are currently being developed as novel antitumor therapeutics. To enhance their therapeutic potential, adenoviruses are being administered in combination with standard chemotherapy. Adenoviral vectors used in these clinical trials, however, can be destructive as they encode intact E1B 19-kDa protein, which can block the apoptotic pathway induced by a variety of chemotherapeutic agents. Previously, we have shown that oncolytic adenovirus Ad-ΔE1B19/55, deleted for sequence encoding E1B 19-kDa and E1B 55-kDa proteins, exhibits marked enhancement in cytolytic and apoptotic activity [Kim, J., Cho, J.Y., Kim, J.H., Jung, K.C., and Yun, C.O. (2002). Cancer Gene Ther. 9, ...

Erythropoietin (Epo), a 30.4-kD glycoprotein, is the principal regulator of erythropoiesis. To evaluate the concept that in vivo gene transfer might be used as an alternative to recombinant human Epo (rhEpo) in applications requiring a 1- to 3-week stimulation of erythropoiesis, the replication-deficient recombinant adenovirus AdMLP.Epo was constructed by deleting the majority of E1 from adenovirus type 5, and replacing E1 with an expression cassette containing the adenovirus type 5 major late promoter (MLP) and the human Epo gene, including the 32 cis-acting hypoxia response element. In vitro studies showed that infection of the human hepatocyte cell line Hep3B with AdMLP.Epo resulted in a 15-fold increase in Epo production in 24 hours that was enhanced to 116-fold in the presence of a hypoxic stimulus. One-time in vivo administration of AdMLP.Epo (7 x 10(9) plaque-forming units/kg) to the peritoneum of cotton rats caused a marked increase in red blood cell production, with a 2.6-fold increase ...

Last year, Chiu and colleagues also identified another new adenovirus, named simian adenovirus C, which sickened four of nine captive baboons and killed two of them at a primate facility in 1997. Several staff members at the facility also complained of upper respiratory symptoms at the time of the outbreak. Re-examining the samples many years later, Chiu and his colleagues found antibodies targeted to simian adenovirus C in the human samples.. Chiu concluded that staff members had been exposed to the new virus, and that the virus may have jumped from baboon to human, an idea also supported by follow-up experiments in which laboratory strains of simian adenovirus C efficiently infected both human and baboon cells.. Adenoviruses to date have not generally been linked to cross-species infections between monkeys and humans, Chiu said.. In light of these findings, however, he said the normal vigilance in tracking animal viruses that might also infect humans should extend beyond influenza and ...

As much as 90% of an intravenously (i.v.) injected dose of adenovirus serotype 5 (Ad5) is absorbed and destroyed by liver Kupffer cells. Viruses that escape these cells can then transduce hepatocytes after binding factor X (FX). Given that interactions with FX and Kupffer cells are thought to occur on the Ad5 hexon protein, we replaced its exposed hypervariable regions (HVR) with those from Ad6. When tested in vivo in BALB/c mice and in hamsters, the Ad5/6 chimera mediated |10 times higher transduction in the liver. This effect was not due to changes in FX binding. Rather, Ad5/6 appeared to escape Kupffer cell uptake as evidenced by producing no Kupffer cell death in vivo, not requiring predosing in vivo, and being phagocytosed less efficiently by macrophages in vitro compared to Ad5. When tested as a helper-dependent adenovirus (Ad) vector, Ad5/6 mediated higher luciferase and factor IX transgene expression than either helper-dependent adenoviral 5 (HD-Ad5) or HD-Ad6 vectors. These data suggest that

Background: A newly demonstrated capability to directly convert fibroblasts into cardiomyocyte-like cells (induced cardiomyocytes [iCMs]) has great potential for the in situ regeneration of functional myocardial tissue from cardiac scar tissue and treatment of cardiovascular disease. Whereas such transdifferentiation has previously been demonstrated using retrovirus and lentivirus vectors, the clinical utility of this cardiac regeneration strategy would be facilitated by the use of a non-integrating, short-term expression viral vector such as adenovirus. In the present study, we sought to determine whether rat cardiac fibroblasts could be converted to iCMs by adenoviral expression of Gata4 (G), Mef2c (M), and Tbx5 (T) and compare the efficiencies of two viral reprogramming strategies.. Methods and Results: Human adenovirus serotype 5 (Ad5) or lentivirus expressing rat GMT or corresponding null viruses were transduced into cultures of rat primary cardiac fibroblasts. After 48 hours viruses were ...

RNA molecules from nuclear and cytoplasmic polyribosomes of adenovirus-infected HeLa cells were compared by hybridization to analyse the sequence content. Nuclear polyribosomes were released by exposure of intact detergent-washed nuclei to poly(U) and purified. Cytoplasmic polyribosomes were also purified from the same cells. To show that nuclear polyribosomes contain ribosomes linked by mRNA, polyribosomes were labelled with methionine and uridine in the presence of actinomycin D in adenovirus-infected cells. Purified nuclear polyribosomes were treated with EDTA under conditions which dissociate polyribosomes into ribosomes and subunits with a simultaneous release of mRNA, and sedimented. The treatment dissociated these polyribosomes, releasing the mRNA from them. Radiolabelled total RNA from each polyribosome population was fractionated in sucrose gradients into several pools or hybridized to intact adenovirus DNA to select virus-specific RNA. Sucrose-gradient-fractionated pool-3 RNA (about ...

TY - JOUR. T1 - Induction of endogenous genes following infection of human endothelial cells with an E1- E4+ adenovirus gene transfer vector. AU - Ramalingam, Ramachandran. AU - Rafii, Shahin. AU - Worgall, Stefan. AU - Hackett, Neil R.. AU - Crystal, Ronald. PY - 1999/12/1. Y1 - 1999/12/1. N2 - Recombinant adenovirus (Ad) gene transfer vectors are effective at transferring exogenous genes to a variety of cells and tissue types both in vitro and in vivo. However, in the process of gene transfer, the Ad vectors induce the expression of target cell genes, some of which may modify the function of the target cell and/or alter the local milieu. To develop a broader understanding of Ad vector-mediated induction of endogenous gene expression, genes induced by first-generation E1- E4+ Ad vectors in primary human umbilical vein endothelial cells were identified by cDNA subtraction cloning. The identified cDNAs included signaling molecules (lymphoid blast crisis [LBC], guanine nucleotide binding-protein ...

Nitric oxide (NO), produced by nitric oxide synthase (NOS), plays key roles in the cardiovascular system, including anti-inflammatory actions. Loss of NOS/NO activity may be important in inflammation and vascular injury in early vein grafts (VG). We used a recombinant adenoviral vector (Ad.nNOS), to augment NOS activity in VG. Methods: Male New Zealand White rabbits (n=16) underwent interposition venous bypass grafting to the carotid artery using the jugular vein. Ad.nNOS (5 × 10 9pfu/ml) was instilled into the VG for 20 minutes during surgery. Experimental vessels and controls were harvested at 3 or 7 days (n=4 all groups). Vessel wet weight /length ratio was measured, and frozen sections analysed for: (1) presence of NOS by NADPH diaphorase (NADPH/d) staining; (2) expression of adhesion molecules (VCAM, ICAM) and (3) inflammatory cell markers (CD18, RAM11) by immunohistochemistry, using computer-aided image analysis; (4) intimal thickness on histology. Results: Adhesion molecule expression,

To generate a replication-competent adenovirus (Ad) with specificity for melanoma, we constructed a tissue-specific promoter restricting E1A expression to melanoma cells. The combination of four copies of a mouse tyrosinase enhancer element (TE) fused to the human tyrosinase promoter (TP) yielded up to 2000-fold higher luciferase reporter activity in tyrosinase-expressing melanoma cells than in nonmelanoma cells. Insertion of the composite TETP construct upstream of the E1A gene was combined with deleting as far as possible the intertwined endogenous Ad enhancer/promoter (EP). The resulting AdDeltaEP-TETP vector, also deleted for the E3 region, was found to replicate in tyrosinase-positive melanoma cells, such as SK-Mel23 as efficiently as wild-type Ad5, but at a more than 50-fold reduced level in nonmelanoma tumour cells and primary human cells. Injection of AdDeltaEP-TETP into xenotransplanted melanomas, but not into HeLa-derived tumours led to long-lasting tumour regression in nude mice. This ...

The utility of recombinant adenovirus serotype 5 (rAd5) vector-based vaccines for HIV-1 and other pathogens will likely be limited by the high prevalence of pre-existing Ad5-specific neutralizing Abs (NAbs) in human populations. However, the immunodominant targets of Ad5-specific NAbs in humans remain poorly characterized. In this study, we assess the titers and primary determinants of Ad5-specific NAbs in individuals from both the United States and the developing world. Importantly, median Ad5-specific NAb titers were ,10-fold higher in sub-Saharan Africa compared with the United States. Moreover, hexon-specific NAb titers were 4- to 10-fold higher than fiber-specific NAb titers in these cohorts by virus neutralization assays using capsid chimeric viruses. We next performed adoptive transfer studies in mice to evaluate the functional capacity of hexon- and fiber-specific NAbs to suppress the immunogenicity of a prototype rAd5-Env vaccine. Hexon-specific NAbs were remarkably efficient at ...

Haidari, M. et al. (2014). Disruption of endothelial adherens junctions by high glucose is mediated by protein kinase C-β-dependent vascular endothelial cadherin tyrosine phosphorylation. Cardiovasc Diabetol. 13:112.. Ackerman, W. et al. (2008). Nuclear Factor-kappa B regulates inducible prostaglandin E synthase expression in human amnion mesenchymal cells. Biol. Reprod. 78:68-76. ...

Infection of mice or rats with adenoviruses can alter their normal immune response and thereby skew experimental data. For example, infection with MAD-1 can produce extensive persistent lesions in the kidneys of adult mice and render them more susceptible to experimental Escherichia coli-induced pyelonephritis. Mouse adenovirus infection has also been shown to accelerate experimental scrapie infection in mice. Although mouse adenoviral infection is usually subclinical in immunocompetent mice, wasting may result in nude mice. ...

Tumor necrosis factor (TNF) alpha has been shown to be a major therapeutic target in rheumatoid arthritis with the success of anti-TNFalpha antibody clinical trials. Although signaling pathways leading to TNFalpha expression have been studied in some detail, there is evidence for considerable differences between individual cell types. This prompted us to investigate the intracellular signaling pathways that result in increased TNFalpha synthesis from macrophages in the diseased synovial joint tissue. Using an adenoviral system in vitro we report the successful delivery of genes to more than 95% of normal human macrophages. This permitted us to show, by using adenoviral transfer of IkappaB alpha, the natural inhibitor of NF-kappaB, that induction of TNFalpha in normal human macrophages by lipopolysaccharide, but not by some other stimuli, was inhibited by 80%. Furthermore the spontaneous production of TNFalpha from human rheumatoid joint cell cultures was inhibited by 75%, indicating that the NF-kappaB

Background: Functional activation of oncogenic K-Ras signaling pathway plays an important role in the early events of colorectal carcinogenesis (CRC). K-Ras proto-oncogene is involved in 35-40% of CRC cases. Mutations in the Ras gene trigger the transduction of proliferative and anti-apoptotic signals, even in the absence of extra cellular stimuli. The objective of the current study was to use a gene-targeting approach to kill human CRC cells selectively harboring mutated K-Ras. Results: A recombinant adenovirus that carries a lethal gene, PUMA, under the control of a Ras responsive promoter (Ad-Py4-SV40-PUMA) was used selectively to target CRC cells (HCT116, SW480, DLD1 and RIE-Ras) that possess a hyperactive Ras pathway while using HT29 and RIE cells as a control that harbors wild type Ras and exhibit very low Ras activity. Control vector, without the Ras responsive promoter elements was used to assess the specificity of our gene therapy approach. Both adenoviral vectors were assed in vitro ...

First-generation, E1-deleted adenoviral vectors (E1-AV) can transduce the vascular endothelium with high efficiency, but their use is limited by the resulting acute endothelial injury and the long-term development of intimal hyperplasia. To reduce the impact of viral proteins on the gene-modified cells, a second-generation adenoviral vector with an additional pair of deletions in the E4 region was developed. To determine whether this E1/E4-AV vector would be useful for vascular gene transfer, we directly compared the efficiency of gene transfer to uninjured rabbit carotid arteries using either an E1/E4-AV or an E1-AV vector encoding beta-galactosidase. Both vectors efficiently transduced vascular endothelium; however, the E1/E4-AV vector gene-modified vessels showed higher beta-galactosidase expression 10 days after gene transfer. Importantly, the E1/E4-AV vector produced substantially less endothelial cell activation, less inflammation, and reduced neointimal hyperplasia compared with the E1-AV vector

This invention is related to adenoviral (Ad) vectors and their applications in the field of genetic medicine, including, but not limited to, gene vaccination, gene transfer, gene therapy, and the like. More specifically, this invention is related to the Ad vectors that carry the minimal cis-element of the Ad genome (minimal Ad vector) and are capable of delivering about 36 kb to about 38 kb of heterologous DNA. The generation and propagation of the minimal Ad vectors require trans-complementation of a packaging-attenuated and replication-defective helper Ad (helper) in an Ad helper cell line. This invention further comprises minimal adenoviral vectors for use in the treatment or prevention of disease or other medical conditions, methodologies for generating such vectors and animal test systems for in vivo evaluation of such Ad vectors. More specifically, this invention describes HIV and/or HPV Ad vectors that contain minimal cis-elements of the Ad genome and comprise HIV and/or HPV nucleic

In vitro experimentation Construction of replication-defective adenoviral vectors. The OPGΔ cDNA was transferred from the mOPGΔ pCEP4 construct into the shuttle vector pTrackCMV using the pAdEasy-1 system ( 27). Recombinant adenoviral plasmids recovered by homologous recombination in Escherichia coli were transfected into 911 cells. Appropriate recombinant plaques containing mOPG were isolated, propagated and titrated. Adenovirus containing GFP was used as a control (Ad-EGFP).. In vivo experimentations Rat osteosarcoma model. The osteosarcoma was initially induced by a local injection of colloidal radioactive 144cerium in rats ( 28). The evolution of the tumor is comparable at the temporal (ratio 1:100 between rats and humans) and physiologic levels to the development of human osteosarcoma. The tumor can be regrafted as described above and maintained in vivo for many months, or fragments can be frozen until reuse. Lung metastases are observed in 75% to 90% of rats bearing advanced malignant ...

TY - JOUR. T1 - EUS or percutaneously guided intratumoral TNFerade biologic with 5-fluorouracil and radiotherapy for first-line treatment of locally advanced pancreatic cancer. T2 - A phase I/II study. AU - Hecht, J. Randolph. AU - Farrell, James J.. AU - Senzer, Neil. AU - Nemunaitis, John. AU - Rosemurgy, Alexander. AU - Chung, Theodore. AU - Hanna, Nader. AU - Chang, Kenneth J.. AU - Javle, Milind. AU - Posner, Mitchell. AU - Waxman, Irving. AU - Reid, Anthony. AU - Erickson, Richard. AU - Canto, Marcia. AU - Chak, Amitabh. AU - Blatner, Gretta. AU - Kovacevic, Milan. AU - Thornton, Mark. N1 - Funding Information: DISCLOSURE: This Phase1/2 study was funded by GenVec, Inc., Gaithersburg, Maryland. No other financial relationships relevant to this publication were disclosed. Copyright: Copyright 2013 Elsevier B.V., All rights reserved.. PY - 2012/2. Y1 - 2012/2. N2 - Background: TNFeradeBiologic (AdGVEGR.TNF.11D) is a replication-deficient adenoviral vector that expresses tumor necrosis ...

TMEM166 is a novel programmed cell death-related molecule. In this report, we constructed a recombinant adenovirus 5-TMEM166 vector (Ad5-TMEM166) and evaluated its expression and anti-tumor activities in vitro and in vivo. Cell viability analysis revealed that the adenovirus-mediated increase of TMEM166 inhibited tumor cell growth in a dose- and time-dependent manner. This inhibitory effect was mediated by both autophagy (via inhibition of mTOR and activation of p70S6K) and apoptosis (via caspase-3 activation), both of which contributed to cell death and suppression of tumorigenicity. Our data indicated that Ad5-TMEM166 may be a novel gene therapy candidate for cancer. (C) 2012 Elsevier Ireland Ltd. All rights reserved. ...

Background. We report the first-in-human safety and immunogenicity assessment of a prototype Ad26 vector-based human immunodeficiency virus (HIV) vaccine in humans. Methods. Sixty Ad26-seronegative, healthy, HIV-uninfected subjects were enrolled in a randomized, double-blinded, placebo-controlled, dose-escalation phase 1 study. Five groups of 12 subjects received 10(9)-10(11) vp of the Ad26-EnvA vaccine (N = 10/group) or placebo (N = 2/group) at weeks 0 and 24 or weeks 0, 4, and 24. Safety and immunogenicity were assessed. Results. Self-limited reactogenicity was observed after the initial immunization at the highest (10(11) vp) dose. No product-related SAEs were observed. All subjects who received the Ad26-EnvA vaccine developed Ad26 NAb titers, EnvA-specific enzyme-linked immunosorbent assays (ELISA) titers, and EnvA-specific enzyme-linked immunospot assays (ELISPOT) responses. These responses persisted at week 52. At week 28 in the 10(9), 10(10), 10(11) vp 3-dose and the 10(10) and 5 × ...

In this study, we used transgenic NF-κB reporter mice and adenoviral vectors to mediate activation and inhibition of NF-κB specifically in lung epithelium. This approach employs novel methodology to study this signal transduction pathway in live animals. Expression of constitutively active forms of IKK1or IKK2 in airway epithelium activates NF-κB with resultant cytokine and chemokine production and neutrophilic lung inflammation. This inflammatory response is blocked by a dominant inhibitor of NF-κB, showing that generation of inflammation by Ad-cIKK1 and Ad-cIKK2 is specifically dependent on NF-κB. Together, these data show that activation of NF-κB in airway epithelium by cIKK1 or cIKK2 is sufficient to produce neutrophilic lung inflammation.. Recombinant adenoviral vectors are efficient vehicles for delivery of genes and are useful for targeting airway epithelium. Ad offer the advantage of high titer, stability, and the ability to achieve gene transfer independent of the cell replication ...

A replication-defective adenoviral vector, Ad.Egr-TNF.11D, was engineered by ligating the CArG (CC(A/T)6GG) elements of the Egr-1 gene promoter upstream to a cDNA encoding human tumor necrosis factor-alpha. We report here that Ad.Egr-TNF.11D is activated by the clinically important anticancer agents cisplatin, cyclophosphamide, doxorubicin, 5-fluorouracil, gemcitabine, and paclitaxel. N-acetylcysteine, a free radical scavenger, blocked induction of tumor necrosis factor-alpha by anticancer agents, supporting a role for reactive oxygen intermediates in activation of the CArG sequences. Importantly, resistance of PC-3 human prostate carcinoma and PROb rat colon carcinoma tumors to doxorubicin in vivo was reversed by combining doxorubicin with Ad.Egr-TNF and resulted in significant antitumor effects. Treatment with Ad.Egr-TNF.11D has been associated with inhibition of tumor angiogenesis. In this context, a significant decrease in tumor microvessel density was observed following combined treatment with

Principal Investigator：KOBAYASHI Atsuki,荒井 勲,阿久根 徹,竹下 克志, Project Period (FY)：1998 - 2001, Research Category：Grant-in-Aid for Scientific Research (C), Section：一般, Research Field：Orthopaedic surgery

In the present study, we showed that gene transfer of rat MCP-1 to the vascular wall of the carotid artery induced early histological changes associated with atherosclerosis in rabbits treated with a high cholesterol diet. Gene transfer of MCP-1 alone in the absence of hypercholesterolemia failed to induce the vascular lesion. The histological changes associated with atherosclerosis were not detected in carotid arteries when rabbits were treated with a high cholesterol diet for 4 weeks in the absence of MCP-1 gene transfer. The vascular lesion developed by MCP-1 gene transfer under hypercholesterolemia is in accordance with fatty streaks or intimal xanthoma, which occurs in the early phase of atherosclerosis.14,15. An increasing amount of evidence has demonstrated the important role of MCP-1 in the initiation and development of atherosclerosis.16,17 In addition to the potent chemoattractant action for circulating monocytes, MCP-1 exerts various effects on monocytes/macrophages. MCP-1 induces the ...

Press Release issued Dec 9, 2014: The gene therapy market has undergone a series of transformation from past few years. Initially gene therapy of monogenetic diseases was popular; however now the gene therapy market is gaining popularity for treatment of cancer. Cancer gene therapy refers to a process of treatment of cancer by inserting therapeutic DNA into patient. Cancer gene therapy is gaining popularity as a result of its success rate in preclinical and clinical trial stages. The most common technique for cancer gene therapy involves replacing a mutated gene that is causing cancer with a healthy copy of gene. Other technique involves inactivation of a gene that is not working properly. A new technique that is being introduced to cancer gene therapy market involves insertion of new genes into the body that would help to fight against tumor cells.

A Human Type 5 Adenovirus-Based Tuberculosis Vaccine Induces Robust T Cell Responses in Humans Despite Preexisting Anti-Adenovirus Immunity Academic Article ...

Enabling topical immunization via microporation: a novel method for pain-free and needle-free delivery of adenovirus-based vaccines Academic Article ...

Adenovirus has been associated with both sporadic and epidemic disease and, with regard to infections among military recruits, who were routinely immunized against types 4 and 7 from 1971 until the cessation of vaccine production in 1996. Adenovirus became a significant cause of economic cost and morbidity in this setting. A live oral vaccine against adenovirus types 4 and 7 was approved for use in this population by the US Food and Drug Administration (FDA) in 2011, and subsequent incidence of acute respiratory disease declined.. Of interest is the role of adenoviruses as vectors in vaccination and in gene therapy. [1, 2, 3] Adenoviruses can infect various cells, both proliferating and quiescent, and thus hold the promise of targeting many different tissues and diseased cell lines.. The genome of adenovirus is well known and can be modified with relative ease to induce lysis or cytotoxicity of a specified cell line without affecting others.. The virus itself can be engineered to remove its ...

Fusion of the BCL9 HD2 domain to E1A increases the cytopathic effect of an oncolytic adenovirus that targets colon cancer cells. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.

BioMed Research International is a peer-reviewed, Open Access journal that publishes original research articles, review articles, and clinical studies covering a wide range of subjects in life sciences and medicine. The journal is divided into 55 subject-specific sections.

Glioblastoma multiforme (GBM) is an aggressive cancer that affects millions of patients per year. Conventional therapies combining chemotherapeutic agents with radiation can only extend survival by a few months; therefore, there is a dire need for an effective means of treating this deadly disease. Melanoma differentiation-associated gene-7/interleukin-24 (mda-7/IL-24), currently in the early stages of FDA pre-IND drug trials, has proven to be an effective cancer specific cytokine, able to trigger the onset of mitochondrial dysfunction and/or autophagy. GBMs have mutations that often result in the activation of cytoprotective cell signaling pathways, preventing cancer therapeutics and even MDA-7/IL-24 treatments from being effective. Since the discovery of MDA-7/IL-24 a number of groups have shown toxic effects in a variety of tumor cells. However, the lethality of MDA-7/IL-24 is not enough to eradicate the tumor. We hypothesized two xxiii rationales for this minimalistic effect. First, the MDA-7/IL-24

Background: Previously we reported that a hexon-modified adenovirus (Ad) vector containing the invasive neutralizing epitope of Trypanosoma cruzi (T. cruzi) trypomastigote gp83 (Ad5-gp83) provided immunoprotection against T. cruzi infection. The purpose of this work was to design an improved vaccine for T. cruzi using a novel epitope capsid incorporation strategy. Thus, we evaluated the immunoprotection raised by co-immunization with Ad5-gp83 and an Ad vector containing an epitope (ASP-M) of the T. cruzi amastigote surface protein 2. Methods: Protein IX (pIX)-modified Ad vector (Ad5-pIX-ASP-M) was generated, characterized, and validated. C3H/He mice were immunized with Ad5-pIX-ASP-M and Ad5-gp83 and the cell-mediated responses were evaluated by enzyme-linked immunospot (ELISPOT) assay and intracellular staining. Immunized mice were challenged with T. cruzi to evaluate the vaccine efficacy. Results: Our findings indicate that Ad5-pIX-ASP-M was viable. Specific CD8+ T-cell mediated responses prior ...

The present invention relates to a genic expression adenoviral hybrid vector characterized in that it contains at least the following elements, oriented in the direction 5 to 3: i. a first chain of adenoviral origin comprising a first inverted terminal repeat (ITR) sequence and a signal sequence for packaging of the adenovirus; ii. a first non-encoding stuffer sequence; iii. a sequence corresponding to a tissue specific promoter; iv. a chain of cDNA derived from an alphavirus, the sequence of which is partly complementary to an alphaviral RNA sequence, comprising at least a sequence encoding for at least one exogenous gene of interest; v. a polyadenylation sequence; and vi. a second adenoviral inverted terminal repeat (ITR) sequence, it preferably relates to an adenoviral hybrid vector comprising as exogenous gene of interest the therapeutic gene of mammalian interleukin IL-12 and even more preferably human interleukin hIL-12; and to the use of the

This is a Phase 1 randomized, single center, dose-escalation study designed to evaluate the safety and immunogenicity of live, replication competent recombinant Adenovirus type 4-H5N1Influenza Vietnam 1194 Hemagglutinin (HA) (Ad4-H5-Vtn). Determining the optimal route and dose for this recombinant platform will greatly accelerate investigations of this vector as an influenza vaccine and an HIV vaccine platform.. Tonsillar administration of the vaccine will be separately assessed. The oral enteric-coated capsule will also be assessed in 10 outpatients using similar blood sampling for comparison. The Ad4-H5-Vtn orally administered as enteric-coated capsules has already been evaluated in an ascending dose Phase 1 study, in dosages as high as 10(11) viral particles (vp).. The primary goal of this study is to evaluate safety of ascending dosages of the Ad4-H5-Vtn vaccine following intranasal and tonsillar administration. A dosage or dosages will be selected to further evaluate the humoral, cellular, ...

Replication deficient adenoviruses type 5 (Ad5) are widely used vectors for cancer gene therapy. The retargeting of Ad5 is achieved by incorporation of a targeting motifs into the adenovirus capsid proteins, mostly in the HI-loop of fiber or hexon hypervariable region 5 (HVR5). Increased expression of αv integrins has been frequently shown in tumor cells as compared to normal cells. The targeting motif within Ad5 that retargets its transduction to cells expressing αv integrins is RGD. One of the strategies of tumor gene therapy is inhibition of angiogenesis. Endothelial cells in angiogenesis express aminopeptidase N (APN) which binds targeting motifs containing NGR. It is known that different amino acid environment and/or cysteine residues flanking targeting peptides RGD and NGR influence the affinity and specificy of binding to corresponding receptors. The aim of this work was to investigate the existance of disulfide bond/s in two replication deficient Ad5 vectors: (i) Ad5HCRGDC containing ...

Malignant glioblastomas and melanomas continue to have a dismal prognosis despite advances in conventional therapy. This has led to investigations of novel treatment strategies including immunogene th

Full custom adenovirus production services, adenovirus vector construction, adenovirus packaging, adenovirus amplification, adenovirus purification, adenovirus titering.

The oncolytic potency of CRAds is critically linked to effective adenoviral infection and replication. First, there must be efficient infection in target cells. To this end, we have shown gains in infectivity via CAR-independent infection to overcome the hurdle that most tumor cells are deficient with respect to the level of the CAR receptor. These gains provide a logical framework for addressing other biological factors related to CRAd potency, such as efficacy of replication, as studied here. Furthermore, elucidation of distinct mechanisms will allow us to develop a strategy to circumvent other barriers and improve the potency of CRAds. In the present study, we endeavored to define the replicative blocks that may be limiting the full potential of oncolytic CRAds.. We showed that immortalized cell lines support wild-type adenoviral replication to a much greater degree than primary patient tumors. Because fiber and hexon are both late proteins involved in the assembly of virions, it was expected ...

On the second day Prof R Geoff Richards gave an illustrative lecture about the importance of pre-clinical translation and demonstrated some recent examples of products and procedures developed by the AO. Although the way from bench to bedside is long and the AO Research Institute Davos is primarily involved in the first steps, translation of research findings into the clinics has always been the ultimate goal for all AO Research Institute Davos projects. The course was completed by different workshops addressing relevant topics from in vitro and in vivo studies for skeletal research to real clinical cases. Adenovirus transduction was demonstrated by Dr Martin Stoddart, adhesive polymers by Dr David Eglin, implant infection by Dr Fintan Moriarty, disc organ culture by Dr Sibylle Grad, in vivo models by Dr Stephan Zeiter, and bone microscopy by Christoph Sprecher. Bernd Heinlein and Daniel Baumgartner from ZHAW led the workshop on endoprosthetics, while Dr Raphael Jenni involved the participants ...

The 293 cell line was derived from primary cultures of human embryonic kidney (HEK) cells with sheared fragments of adenovirus (Ad) 5 DNA (Graham et al. 1977). HEK 293 cells contain the nucleotides 1-4344 of Ad5 which are located within the pregnancy-specific ß-1-glycoprotein 4 (PSG 4) gene. The transforming region of the human adenovirus contains the early region (E1), comprising two transcription units, E1a and E1b, whose products are essential and sufficient for mammalian cell transformation by adenoviruses (Louis et al. 1997). Because 293 cells express E1 gene products they are extensively used for the production of E1-deleted Ad viruses. Adeno-associated viruses (AAVs) belong to the family of Parvoviridae, being one of the smallest single-stranded and non-enveloped DNA viruses. AAVss are replication-deficient and have required co-infection with a helper adeno- or herpes virus for productive infection. The AAV Helper-free system takes advantage of the identification of the specific ...

Vector construction. Construction of recombinant adenoviral vectors expressing human IL12 under control of doxycycline by calcium phosphate-mediated coprecipita