adenovirus e1 proteins

..6 kb) was exploited to construct a vector encoding the entire hepatitis C virus (HCV) polyprotein. Infection of HeLa cells by the resulting vector showed high levels of correctly processed HCV proteins...

..Our novel Ad35-based vectors and their corresponding packaging cell lines will provide a versatile and powerful system for DNA-based vaccine development and gene therapy applications...

..inflammatory responses by upregulating expression of adhesion molecules and enhancing migration through Advector-infected ECs and suggest that E1(-)E4(-) Advectors may be a better choice for gene-transfer strategies directed to the ECS:..

....

..Since the HSV amplicon can be easily and completely inactivated by chloroform extraction and/or heat treatment from the HDAdV stock, this helper system might be an alternative method to produce helper virus-free HDAdVs...

..Taken together our results suggest that the binary vectors described here should be a valuable addition to the repertoire of viral vectors used in basic and medical research...

..Gene expression changes induced by an E1/E3-defective adenovirus vector have been studied in human mammary epithelial cells by comparing the gene expression profile in infected and uninfected cells...

..These results highlight the possibility of using low level infection with recombinant adenovirus to cause short-term blocks on cell proliferation...

..This novel cloning strategy allows the rapid and versatile construction of adenoviral vectors for gene expression and gene therapy applications...

..However, the response elicited by the AdHu5 vaccine is greatly attenuated in AdHu5 preimmune animals...

..They are equivalent to sequential vaccinations with an E1-deleted Ad vector followed by booster immunization with a poxvirus vector and they surpass regimens based on DNA vaccine prime followed by a recombinant adenoviral vector boost...

..These data support the potential utility of TNFR-Ig gene transfer to islets as a means of facilitating allogeneic islet transplantation...

..The development of somatic cell hybrids appears to be a simple way of combining some of the desirable characteristics present separately in two parental cell lines...

....

..The combination of CRAD and E1-deleted ad induced tumor-specific replication of CRAD and E1-deleted ad and increased the transduction rate and therapeutic efficacy of these viruses in model tumors...

..Helper-dependent adenoviral vectors will be useful agents for vascular gene transfer and gene therapy...

..98% without compromising hybrid vector rescue. This allowed amplification of high-capacity AAV/Ad hybrid vectors to high titers in a single round of propagation...

..These results were validated in a 3 L bioreactor under controlled operating conditions where a higher HDAd yield of 2.6 x 10(9) viral particles (VP)/mL or 3.5 x 10(8) infectious units (IU)/mL of HDAd was obtained...

..We demonstrated that PAV219 was able to transduce and direct expression of GFP in some human cell lines...

..The prostate-targeted gene delivery vectors with robust and cell-specific expression capability will advance the development of safe and effective imaging guided therapy for recurrent metastatic stages of prostate cancer...

....

..These data clearly demonstrate that CRAd-Survivin-RGD is a potential novel therapeutic agent for treatment in many, but not all, human cancers...

..In addition, this technique has potential utility for dynamic monitoring of adenovirus replication and spread...

..These results have important implications for the use of genetically modified DC for therapeutic application...

..C6 cells were analysed and found to be free of RCA and HDEP. Based on the statistical model used, we anticipate that our approach to RCA assay development can be broadly applicable to other adenoviral vectors...

..Given the well-documented immunogenicity of the first-generation adenoviral vector, data from C57BL/6 mice may therefore grossly underestimate immunological consequences in certain gene therapy protocols...

..These results add to the evidence that cell-mediated immunity is a substantial barrier to therapeutic adenoviral vectors and provide more quantitative tools to measure cellular immune responses to adenovirus...

..Increased infectivity and altered tropism were noted in the treated tumor tissue. Taken together, this ex vivo system indicated that receptor utilization and extracellular-matrix components influence AD viral tropism in solid tissues...

..This method should be useful for RNA interference-based experiments, and should make it easy to construct an siRNA-expressing Ad vector library for functional screening...

..CONCLUSIONS: Our results suggest that the hTERT promoter confers competence for selective replication of TRAD in human cancer cells, an outcome that has important implications for the treatment of human cancers...

..Nevertheless, mice remain responsive to the transgene product delivered by a viral vector...

....

..On the basis of a careful review of the literature, these improvements in vector safety rival those noted with other, more significantly modified Ad vectors described to date...

..These results indicate that chimpanzee adenoviral vectors may be useful as vaccines or gene therapy vectors in human populations and should allow applications that require multiple vector administrations...

..This finding allowed development of an HDEP assay to screen batches of rAd5 produced on the PER.C6 cell line and resulted in detection of seven HDEP agents from four different transgene-virus vector constructs in separate batches of Ad...

..Therefore, administration of ADV5-based vectors in DNA-primed subjects may be a preferred use of this vaccine modality for generating long-term immune protection...

..The structure of the viral genome and life cycle of Ad35 differs from the better characterized Ad5 and these differences require differences in the strategies for the generation of vectors for gene delivery...

..These pre-clinical studies with E1 and E2b-deleted recombinant Ad5 vectors suggest that anti-Ad immunity will no longer be a limiting factor, and that clinical trials to evaluate their performance are warranted...

..In spite of their usefulness, the construction of adenoviral vectors (AdV) is a cumbersome and lengthy process that is not readily amenable to the generation of large collection of clones...

..Knowing that adenoviral vectors could initiate innate immunity, the ability of E1-deleted recombinant adenovirus (Ad-E1Delta) in induction of B7.1 and IL-2 molecules was studied...

..These agents should provide important insights into the identification of novel therapeutic strategies for mesothelioma...

..It is further shown that this strategy may be more widely applicable...

..Presently the E1 and E4 regions are modified in a time consuming sequential fashion...

..Because of this tumor grade-based differential growth, the E1-deleted adenoviruses may represent novel tools for studies of brain tumor malignancy...

..These findings are of value for further molecular studies of cochlear inner hair cells and for gene replacement strategies to correct recessive genetic hearing loss due to monogenic mutations in these cells...

..These results indicate the construction of an optimized Ad35 vector backbone for efficient transduction into HSCs/progenitors...

..Here we report methods for evaluation of contaminating adenovirus and E1 viral DNA...

..Virus neutralizing antibody titers can be enhanced by prime-boost regimens...

..This flexible cloning system allows for straightforward construction of tropism expanded replicating adenoviral vectors that express transgenes late in the adenoviral life cycle...

....

..Future studies of these factors will undoubtedly lead to development of new therapeutic approaches to PR- and RPE-specific retinal diseases...

We have examined the effect of adenovirus E1 proteins on expression from the immediate early (IE) region of the human cytomegalovirus (HCMV)...

....

....

..Gene therapy has been successfully applied to the cochlea of a laboratory animal, and future studies will define the types of vectors and therapeutic genes that will work best for the treatment of inner ear diseases in the clinic...

..In this study, the influence of the E4 adenoviral region on the activity of various promoters was investigated in vitro and in vivo...

..These experiments demonstrate that this treatment is transient and antigen-specific...

....

..These results may have a significant impact on the development of packaging cell lines for replication-deficient adenovirus production...

..We concluded that application of this replication-competent Ad to hematopoietic grafts could be a simple but effective procedure to achieve complete tumor cell purging...

..Overexpression of full-length FLASH blocks the anti-apoptotic effect of the adenovirus protein E1B19K. FLASH is therefore necessary for the activation of caspase-8 in Fas-mediated apoptosis...

..This regulatable adenoviral vector system should be of value in behavioral studies and in vivo studies of neuronal gene function, and may further the development of effective gene therapy strategies in the brain...

..The normalization also produced cells that divided 2 - 5 times and died and cells that reverted to a transformed phenotype in 2 - 10 days. The latter appeared predominantly upon the action of the antisense oligonucleotides...

..Additionally, humoral immune response to transgene products has been demonstrated in immunocompetent animal models, which will also need to be surmounted for long-term efficacy in disease treatment by gene therapy...

..The data emphasize that this ovine virus differs significantly from other known human and animal adenoviruses...

..The potential of this procedure is demonstrated by the recovery of several E1E3E4-deleted adenoviruses following transfection of the corresponding E. coli-derived genomes in IGRP2 cells...

..In contrast, there was no repression in the presence of the regulator tetracycline. We propose that regulation by TetR-KRAB is a valuable tool with which to study the effects of viral gene expression in vitro...

..The results suggested that these modified vectors may be very useful both for in vitro and in vivo gene therapy applications...

..In addition, our data confirm that, in the absence of transgene expression, mouse cellular immunity to viral antigens plays a minor role in the progressive elimination of the virus genome...

..At high doses of vector, widespread dissemination of vector DNA is seen. At low doses, hepatic gene transfer is not associated with vector DNA dissemination to gonads...

..Taken together, these observations demonstrate that CD4(+) T cells are able to eradicate MHC class II-negative tumors in an immune-privileged site without affecting surrounding tissues or the induction of phthisis...

..This effect can be complicated by the action of transgene expression on cell survival...

..Taken together, these results suggest that AUP1 is a cytoplasmic protein that is expressed in all cell lines we examined...

..The present experiment demonstrates that intraneural injection of colchicine is capable of inhibiting remote delivery of both adenoviral and adeno-associated viral (AAV) vectors, implicating axonal transport in this process...

..These results suggest that the 2.7 kb of MoMLV sequence included in AdELP-luc have cis-acting functions and mediates an unusual integration event...

..Thus, we constructed infectivity enhanced cyclooxygenase (COX)-2 promoter-based CRAds to develop a safe and effective therapeutic modality...

..The efficacy of the AdHu5rab.gp vector given orally to newborn mice born to AdHu5 virus-immune dams was not impaired by maternally transferred Abs to the vaccine carrier...

....

..These results suggest that the Ad5-hTERT-E1 vector has potential as a broad-spectrum antitumor agent...

..This AdLibrary approach provides an opportunity to exploit the efficient gene delivery capabilities of adenovirus vectors for the rapid discovery of gene and protein function...

..These new Ad.IR-E1A vectors combine elements that allow for tumor-specific transgene expression, efficient viral replication, and spread in liver metastases after systemic vector application...

....

..This approach could also be applied to decrease reincorporation of the E1 genes during amplification of deltaE1deltaE4 vectors in 293/E4-trans-complementing cells...

....

..Moreover, Huh7-E1 tumors release human alpha-1-antitrypsin (hAAT), which allows the monitoring of occult growing tumors (i.e. liver, peritoneum) by measuring serum hAAT levels...

..To prevent vector mobilization by wt Ads, new vectors should be developed in which essential functions such as the initiation of DNA replication and genome packaging are restricted...

..These cell lines will be useful in generating and growing BAV3 E1-deleted recombinants, and also for studying E1 protein interactions with a number of cellular and/or viral proteins...

..7K, as expected. The RID-expressing vectors forced the internalization and degradation of Fas and EGFR, and they inhibited apoptosis induced through the Fas pathway. These vectors should be useful reagents to study the E3 proteins...

..This article reviews recent highlights in this rapidly evolving field: cancer therapeutic and cancer diagnostic approaches using the telomerase-specific oncolytic adenoviruses...

..C6 (lacking the 177 nucleotide region of homology) has allowed extensive virus propagation (Ad5.1FGF-4) with no CPE- or HDEP-like events yet detected...

..Further understanding of the transcription initiation activity of the left-end sequence of the adenovirus genome should lead to optimization of adenovirus vectors...

..These studies will lead to improved strategies for re-establishing immune privilege of corneal transplants and offsetting the untoward effects of allergic diseases on the fate of corneal transplants. ..

..abstract_text> ..

..In aim 4, we will determine the tropism of the simian and human Ad recombinants upon oral application emphasizing localization of and cytokine production by transduced antigen presenting cells (APCs). ..

..g., corneal transplantation). In some cases, it may be desirable to abrogate immune privilege to promote the immune-rejection of sight- and life-threatening ocular neoplasms. ..

..g., corneal transplantation) or abrogate immune privilege to protect against sight- and life-threatening neoplasms. ..

..Both strategies procure antigen-specific suppression of host immune responses and should prove useful in preventing corneal graft rejection. ..

....

..g., corneal transplantation). In some cases, it may be desirable to abrogate immune privilege to promote the immune-rejection of sight- and life-threatening ocular neoplasms. ..

..In aim 2, the AdC68rab.gp vaccine will be tested in a non-human primate rabies virus challenge model. ..

....

..The fourth aim will evaluate the therapeutic efficacy of a monoclonal antibody directed against human CD54 in controlling liver metastases in a nude mouse model of uveal melanoma. ..

....

..These studies will lead to improved strategies for re-establishing immune privilege of corneal transplants and offsetting the untoward effects of allergic diseases on the fate of corneal transplants. ..

....

..The projects will be supported by an Administrative Core, which will provide the needed oversight, and a Vector Core, which will provide the investigators with purified and quality controlled vectors. ..

..All three of these aims will be addressed in a well-characterized mouse model of penetrating keratoplasty. ..

..In aim 3, we are proposing experiments in non-human primates to test the vaccine for efficacy in pre- and post-challenge vaccination experiments with and without concomitant application of RIG. ..

..The long range goals of this project are to understand immune regulatory processes in the eye and to manipulate such processes to reduce immune-mediated injury and corneal graft rejection...

..The results from these investigations should provide important insights into the immunobiology of corneal allografts and the feasibility of specific immunological strategies for promoting corneal allograft survival...

..The role of the two different memory CD8+ T cell populations in limiting propagation of the vaccinia gag recombinant virus will be tested by adoptive transfer experiments. ..

..In case the applicant's hypothesis is correct, data gathered within the realm of this application will serve as the basis for a clinical Phase I trial. ..

..All of the course work will take place at The Wistar Institute. ..

..In the third aim, we will test a prime-boost regimen using DNA vaccines, expressing Gag for systemic priming combined with the adenoviral recombinants for systemic or intranasal booster immunization. ..