• Cell therapy
• Gene therapy
• NAFLD
• Non-Alcoholic Fatty Liver Disease
• Stem cell
• Viral vectors

• Humans
• Non-alcoholic Fatty Liver Disease/therapy
• Non-alcoholic Fatty Liver Disease/genetics
• Non-alcoholic Fatty Liver Disease/immunology
• Genetic Therapy/methods
• Hepatocytes/metabolism
• Hepatocytes/cytology
• Animals
• Cell- and Tissue-Based Therapy/methods
• Endoplasmic Reticulum Stress
• Hepatic Stellate Cells/metabolism
• Liver/pathology

• Acute kidney injury
• Platelets
• Self-assembling nanoparticles
• Transformation-related protein 53

• RNA, Small Interfering/chemistry
• RNA, Small Interfering/administration & dosage
• RNA, Small Interfering/pharmacology
• Acute Kidney Injury/therapy
• Blood Platelets/metabolism
• Blood Platelets/chemistry
• Animals
• Humans
• Nanoparticles/chemistry
• Blood Proteins/chemistry
• Tumor Suppressor Protein p53/genetics
• Tumor Suppressor Protein p53/metabolism
• Extracellular Vesicles/metabolism
• Mice
• Male

• 31801088 National Natural Science Foundation of China

• Chronic kidney disease
• Platelet membrane-camouflaging nanoparticles
• Renal fibrosis
• TGF-β1 siRNA
• Targeted therapy

• Animals
• RNA, Small Interfering/administration & dosage
• Transforming Growth Factor beta1/metabolism
• Fibrosis
• Nanoparticles
• Male
• Mice
• Blood Platelets/metabolism
• Reperfusion Injury/drug therapy
• Reperfusion Injury/prevention & control
• Kidney/metabolism
• Kidney/pathology
• Kidney/drug effects
• Mice, Inbred C57BL
• Ureteral Obstruction/therapy
• Biomimetic Materials/administration & dosage
• Biomimetic Materials/chemistry
• Inflammation/drug therapy
• Disease Models, Animal
• Humans
• Smad3 Protein/metabolism
• Polylactic Acid-Polyglycolic Acid Copolymer/chemistry
• Acute Kidney Injury/prevention & control

• adenovirus serotypes 5 and 43 chimera
• hexon hypervariable regions
• high-grade glioma
• neutralizing antibody
• oncolytic adenovirus

• Humans
• Animals
• Mice
• Adenoviridae/genetics
• Antibodies, Neutralizing
• Glioma/therapy
• Glioma/pathology
• Brain Neoplasms/pathology
• Oncolytic Viruses/genetics
• Oncolytic Virotherapy
• Antibodies, Viral
• Oligopeptides/therapeutic use

• P30 CA016672 NCI NIH HHS
• P50 CA127001 NCI NIH HHS
• R01 CA256006 NCI NIH HHS

• Myocarditis
• immune checkpoint inhibitors
• immunotherapy
• mRNA vaccines
• vaccine
• virus

The incidence of myocardial ischaemia‒reperfusion injury (MIRI) is increasing every year, and there is an urgent need to develop new therapeutic approaches. Nrf2 is thought to play a protective role during MIRI and it is regulated by microRNAs (miRNAs). This study focused on PLGA nanoparticles camouflaged by platelet membrane vesicles (PMVs) (i.e., PMVs@PLGA complexes) carrying microRNA inhibitors, which regulate Nrf2 and can play a therapeutic role in the MIRI process. In vitro and in vivo characterization showed that PMVs@PLGA has excellent transfection efficiency, low toxicity and good targeting. MicroRNAs that effectively regulate Nrf2 were identified, and then PMVs@PLGA-miRNA complexes were prepared and used for in vitro and in vivo treatment. PMVs@PLGA-miRNA complexes can effectively target the delivery of inhibitors to cardiomyocytes. Our results suggest that PMVs@PLGA complexes are a novel delivery system and a novel biological approach to the treatment of MIRI.

• Delivery system
• Myocardial ischaemia‒reperfusion injury
• Nrf2
• PLGA
• Platelet membrane vesicles

• PPRV
• adenovirus
• immune response
• vaccines
• viral vector

• Immunocontraceptive
• Infertility
• Naked plasmid DNA
• Recombinant adenovirus
• ZP3

• adenovirus
• cancer
• gene therapy
• oncolytic virus
• vector

• PRRSV
• Protective effect
• Recombinant adenoviruses
• Soluble viral receptors

• Friend virus
• adenovirus
• immunization
• immunodominance
• retrovirus
• vaccine
• vector

Vaccination is one of the most efficient tools for disease prevention, and a continuously growing field of research. However, despite progress, we still need more efficient and cost-effective vaccines that would improve access to those in need. In this review, we will describe the status of virus-vectored vaccine technology with a focus on adenoviral-based vaccines. Adenovirus (Ad) vaccines have proven to be efficient in military vaccinations against Ad4 and Ad7 and as highly efficient vectored vaccines against rabies. The question of how other adenovirus-based vaccines can become as efficient as the rabies vaccine is the underlying theme in this review. Here, we will first give an overview of the basic properties of vectored vaccines, followed by an introduction to the characteristics of adenoviral vectors and previously tested modifications of the vector backbone and expression cassettes, with a focus on how they can contribute to increased vaccine cost-effectiveness. Finally, we will highlight a few successful examples of research that have attempted to improve the use of adenoviral-based vaccines by improving the transgene immunogenicity.

• adenovirus
• affordable vaccine candidates
• immune responses
• protective immunity
• replication competent
• transgene
• vaccine
• vaccine technology platform

• Adenoviridae/genetics
• Animals
• Biotechnology
• Cost-Benefit Analysis
• Genetic Vectors/genetics
• Humans
• Immunity
• Transgenes/genetics
• Transgenes/immunology
• Vaccines, Synthetic/administration & dosage
• Vaccines, Synthetic/genetics
• Vaccines, Synthetic/immunology
• Virus Replication

Human adenovirus 7 (HAdV-7) was responsible for a significant number of fatalities during the 2011 community outbreak in Taiwan. The mechanisms underlying the pathogenesis of severe adenovirus infections in non-immunocompromised individuals remain unclear. Adenovirus pneumonia was associated with pleural effusion in a number of patients from the 2011 outbreak suggesting that similar to bacterial pneumonia, patients diagnosed with adenovirus pneumonia who have pleural effusion are more severely and systemically infected, and may have a more protracted disease course. We hypothesized that the host immunological response determines the severity of adenoviral infection.

This retrospective case series study included patients diagnosed with severe lower respiratory tract infections at the National Cheng Kung University Hospital in southern Taiwan between December 2010 and October 2011. The main inclusion criteria were 1) presence of multifocal patchy infiltrates, lobar consolidation or reticular interstitial opacities in chest X-rays, and 2) presence of adenovirus isolated from respiratory specimens. All patients had adenovirus isolated from respiratory specimens, and were negative for other viruses. Pleural effusion was confirmed in all patients using chest echography. Clinical features and laboratory data were compared in patients with (n = 12) and without (n = 15) parapneumonic effusion.

Presence of parapneumonic effusion was significantly associated with a longer febrile duration, more complicated clinical management, and a greater risk of extrapulmonary involvement, notably hepatitis. Patients without pleural effusion had significantly higher numbers of WBCs, platelets, and absolute segment cell counts (ASCs) compared to patients with pleural effusion (all p < 0.05). Patients without pleural effusion had significantly higher counts of CD4+, CD8+, and CD20+ T cells (all p < 0.05) compared to patients with pleural effusion.

Our data indicated that presence of parapneumonic effusion in adenoviral pneumonia was associated with longer febrile duration, more complicated clinical management, a greater risk of hepatitis, and suppression of host cellular immunity. Further prospective, large-scale studies are needed to validate our results.

• Human adenovirus
• Immune response
• Pleural effusion
• Pneumonia
• T cells

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.

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.

Our findings indicate that Ad5-pIX-ASP-M was viable. Specific CD8⁺ T-cell mediated responses prior to the challenge show an increase in IFNγ and TNFα production. A single immunization with Ad5-pIX-ASP-M provided protection from T. cruzi infection, but co-immunizations with Ad5-pIX-ASP-M and Ad5-gp83 provided a higher immunoprotection and increased survival rate of mice.

Overall, these results suggest that the combination of gp83 and ASP-M specific epitopes onto the capsid-incorporated adenoviruses would provide superior protection against Chagas disease as compared with Ad5-gp83 alone.

• Chagas disease
• T. cruzi vaccine constructs
• amastigote surface protein 2 epitope
• co-immunization
• epitope-capsid incorporation
• immunoprotection
• trypomastigote gp83 neutralizing epitope

Due to the increasing amount of people afflicted worldwide with Chagas disease and an increasing prevalence in the United States, there is a greater need to develop a safe and effective vaccine for this neglected disease. Adenovirus serotype 5 (Ad5) is the most common adenovirus vector used for gene therapy and vaccine approaches, but its efficacy is limited by preexisting vector immunity in humans resulting from natural infections. Therefore, we have employed rare serotype adenovirus 48 (Ad48) as an alternative choice for adenovirus/Chagas vaccine therapy. In this study, we modified Ad5 and Ad48 vectors to contain T. cruzi’s amastigote surface protein 2 (ASP-2) in the adenoviral early gene. We also modified Ad5 and Ad48 vectors to utilize the “Antigen Capsid-Incorporation” strategy by adding T. cruzi epitopes to protein IX (pIX). Mice that were immunized with the modified vectors were able to elicit T. cruzi-specific humoral and cellular responses. This study indicates that Ad48-modified vectors function comparable to or even premium to Ad5-modified vectors. This study provides novel data demonstrating that Ad48 can be used as a potential adenovirus vaccine vector against Chagas disease.

• Chagas’ disease
• Trypanosoma cruzi
• adenovirus serotype 48
• amastigote surface protein-2
• antigen capsid-incorporation
• gp83
• vaccine

The gene knockdown activity of small interfering RNA (siRNA) has led to their use as potential therapeutics for a variety of diseases. However, successful gene therapy requires safe and efficient delivery systems. In this study, we choose mPEG-PLGA-PLL nanoparticles (PEAL NPs) with ultrasound targeted microbubble destruction (UTMD) to efficiently deliver siRNA into cells. An emulsification-solvent evaporation method was used to prepare siRNA-loaded PEAL NPs. The NPs possessed an average size of 132.6±10.3 nm (n=5), with a uniform spherical shape, and had an encapsulation efficiency (EE) of more than 98%. As demonstrated by MTT assay, neither PEAL NPs nor siRNA-loaded PEAL NPs showed cytotoxicity even at high concentrations. The results of cellular uptake showed, with the assistance of UTMD, the siRNA-loaded PEAL NPs can be effectively internalized and can subsequently release siRNA in cells. Taken together, PEAL NPs with UTMD may be highly promising for siRNA delivery, making it possible to fully exploit the potential of siRNA-based therapeutics.

• UTMD
• emulsification-solvent evaporation method
• gene delivery
• mPEG-PLGA-PLL
• orthogonal design

In a recent genome-wide association study, WD-repeat domain 12 (WDR12) was associated with early-onset myocardial infarction (MI). However, the function of WDR12 in the heart is unknown.

We characterized cardiac expression of WDR12, used adenovirus-mediated WDR12 gene delivery to examine effects of WDR12 on left ventricular (LV) remodeling, and analyzed relationship between MI associated WDR12 allele and cardiac function in human subjects. LV WDR12 protein levels were increased in patients with dilated cardiomyopathy and rats post-infarction. In normal adult rat hearts, WDR12 gene delivery into the anterior wall of the LV decreased interventricular septum diastolic and systolic thickness and increased the diastolic and systolic diameters of the LV. Moreover, LV ejection fraction (9.1%, P<0.05) and fractional shortening (12.2%, P<0.05) were declined. The adverse effects of WDR12 gene delivery on cardiac function were associated with decreased cellular proliferation, activation of p38 mitogen–activated protein kinase (MAPK)/heat shock protein (HSP) 27 pathway, and increased protein levels of Block of proliferation 1 (BOP1), essential for ribosome biogenesis. Post-infarction WDR12 gene delivery decreased E/A ratio (32%, P<0.05) suggesting worsening of diastolic function. In human subjects, MI associated WDR12 allele was associated significantly with diastolic dysfunction and left atrial size.

WDR12 triggers distinct deterioration of cardiac function in adult rat heart and the MI associated WDR12 variant is associated with diastolic dysfunction in human subjects.

• Adult
• Alleles
• Animals
• Cell Cycle Proteins
• Cells, Cultured
• Female
• HSP27 Heat-Shock Proteins/genetics
• HSP27 Heat-Shock Proteins/metabolism
• Heart Failure/genetics
• Heart Failure/metabolism
• Heart Failure/physiopathology
• Hemodynamics
• Humans
• Male
• Middle Aged
• Myocardial Infarction/genetics
• Myocardial Infarction/metabolism
• Myocardial Infarction/physiopathology
• Myocytes, Cardiac/metabolism
• Nuclear Proteins/genetics
• Nuclear Proteins/metabolism
• RNA-Binding Proteins
• Rats
• Rats, Sprague-Dawley
• Up-Regulation
• p38 Mitogen-Activated Protein Kinases/genetics
• p38 Mitogen-Activated Protein Kinases/metabolism

In recent years, the increasing prevalence of obesity and obesity-related co-morbidities fostered intensive research in the field of adipose tissue biology. To further unravel molecular mechanisms of adipose tissue function, genetic tools enabling functional studies in vitro and in vivo are essential. While the use of transgenic animals is well established, attempts using viral and non-viral vectors to genetically modify adipocytes in vivo are rare. Therefore, we here characterized recombinant Adeno-associated virus (rAAV) vectors regarding their potency as gene transfer vehicles for adipose tissue. Our results demonstrate that a single dose of systemically applied rAAV8-CMV-eGFP can give rise to remarkable transgene expression in murine adipose tissues. Upon transcriptional targeting of the rAAV8 vector to adipocytes using a 2.2 kb fragment of the murine adiponectin (mAP2.2) promoter, eGFP expression was significantly decreased in off-target tissues while efficient transduction was maintained in subcutaneous and visceral fat depots. Moreover, rAAV8-mAP2.2-mediated expression of perilipin A – a lipid-droplet-associated protein – resulted in significant changes in metabolic parameters only three weeks post vector administration. Taken together, our findings indicate that rAAV vector technology is applicable as a flexible tool to genetically modify adipocytes for functional proof-of-concept studies and the assessment of putative therapeutic targets in vivo.

• Adiponectin/genetics
• Adipose Tissue/virology
• Animals
• Carrier Proteins/genetics
• Carrier Proteins/metabolism
• Dependovirus/genetics
• Gene Transfer Techniques
• Genetic Therapy/methods
• Genetic Vectors
• Green Fluorescent Proteins/analysis
• Mice
• Perilipin-1
• Phosphoproteins/genetics
• Phosphoproteins/metabolism
• Promoter Regions, Genetic

To evaluate localization and transgene expression from adenoviral vector of serotypes 5, 35, and 28, ± an RGD motif in the fiber following intravitreal or subretinal administration.

Ocular transduction by adenoviral vector serotypes ± RGD was studied in the eyes of mice receiving an intravitreous or subretinal injection. Each serotype expressed a CMV-GFP expression cassette and histological sections of eyes were examined. Transgene expression levels were examined using luciferase (Luc) regulated by the CMV promoter.

GFP localization studies revealed that serotypes 5 and 28 given intravitreously transduced corneal endothelial, trabecular, and iris cells. Intravitreous delivery of the unmodified Ad35 serotype transduced only trabecular meshwork cells, but, the modification of the RGD motif into the fiber of the Ad35 viral vector base expanded transduction to corneal endothelial and iris cells. Incorporation of the RGD motif into the fiber knob with deletion of RGD from the penton base did not affect the transduction ability of the Ad5 vector base. Subretinal studies showed that RGD in the Ad5 knob shifted transduction from RPE cells to photoreceptor cells. Using a CMV-Luc expression cassette, intravitreous delivery of all the tested vectors, such as Ad5-, Ad35- and Ad28- resulted in an initial rapid induction of luciferase activity that thereafter declined. Subretinal administration of vectors showed a marked difference in transgene activity. Ad35-Luc gene expression peaked at 7 days and remained elevated for 6 months. Ad28-Luc expression was high after 1 day and remained sustained for one month.

Different adenoviral vector serotypes ± modifications transduce different cells within the eye. Transgene expression can be brief or extended and is serotype and delivery route dependent. Thus, adenoviral vectors provide a versatile platform for the delivery of therapeutic agents for ocular diseases.

• Adenoviridae/genetics
• Animals
• Female
• Gene Expression
• Green Fluorescent Proteins/biosynthesis
• Green Fluorescent Proteins/genetics
• Injections, Intraocular
• Mice, Inbred C57BL
• Oligopeptides/genetics
• Transduction, Genetic

• Adenoviridae/genetics
• Animals
• Capsid Proteins/genetics
• Capsid Proteins/immunology
• Chagas Disease/immunology
• Chagas Disease/prevention & control
• Female
• Genetic Vectors/genetics
• HEK293 Cells
• Humans
• Mice
• Mice, Inbred C57BL
• Protozoan Proteins/genetics
• Protozoan Proteins/immunology
• Trypanosoma cruzi/genetics
• Trypanosoma cruzi/immunology
• Variant Surface Glycoproteins, Trypanosoma/genetics
• Variant Surface Glycoproteins, Trypanosoma/immunology

• T32 HL007737 NHLBI NIH HHS
• U54 CA163069 NCI NIH HHS
• G12 MD007586 NIMHD NIH HHS
• SC2 AI083925 NIAID NIH HHS
• T32 AI007281 NIAID NIH HHS
• T32 AI007493 NIAID NIH HHS
• AI083925 NIAID NIH HHS
• MD007593 NIMHD NIH HHS
• R25 GM059994 NIGMS NIH HHS
• AI080580 NIAID NIH HHS
• AI007281 NIAID NIH HHS
• 5T32AI7493-18 NIAID NIH HHS
• U54 CA163072 NCI NIH HHS
• U54 MD007586 NIMHD NIH HHS
• SC1 AI080580 NIAID NIH HHS
• U54 CA163066 NCI NIH HHS
• MD007586 NIMHD NIH HHS
• R01 AI089337 NIAID NIH HHS
• 5R01AI089337-04 NIAID NIH HHS
• GM059994 NIGMS NIH HHS
• U54 MD007593 NIMHD NIH HHS

MicroRNAs are small (18–22 nucleotide long) noncoding RNAs that play important roles in biological processes through posttranscriptional regulation of gene expression. Their aberrant expression and functional significance are reported in several human malignancies, including pancreatic cancer. Recently, we identified miR-150 as a novel tumor suppressor microRNA in pancreatic cancer. Furthermore, expression of miR-150 was downregulated in the majority of tumor cases, suggesting that its restoration could serve as an effective approach for pancreatic cancer therapy. In the present study, we developed a nanoparticle-based miR-150 delivery system and tested its therapeutic efficacy in vitro. Using double emulsion solvent evaporation method, we developed a poly (D,L-lactide-co-glycolide) (PLGA)-based nanoformulation of miR-150 (miR-150-NF). Polyethyleneimine (a cationic polymer) was incorporated in PLGA matrix to increase the encapsulation of miR-150. Physical characterization of miR-150-NF demonstrated that these nanoparticles had high encapsulation efficiency (~78%) and exhibited sustained release profile. Treatment of pancreatic cancer cells with miR-150-NF led to efficient intracellular delivery of miR-150 mimics and caused significant downregulation of its target gene (MUC4) expression. Inhibition of MUC4 correlated with a concomitant decrease in the expression of its interacting partner, HER2, and repression of its downstream signaling. Furthermore, treatment of pancreatic cancer cells with miR-150-NF suppressed their growth, clonogenicity, motility, and invasion. Together, these findings suggest that PLGA-based nanoformulation could potentially serve as a safe and effective nanovector platform for miR-150 delivery to pancreatic tumor cells.

• MUC4
• PLGA nanoparticles
• invasion
• miR-150
• migration

• kidney
• renal gene delivery
• recombinant adeno-associated virus
• glomeruli
• tubules

• Animals
• Dependovirus/genetics
• Genetic Therapy/methods
• Genetic Vectors/administration & dosage
• Injections
• Kidney/drug effects
• Kidney/physiology
• Mice, Inbred C57BL
• Promoter Regions, Genetic
• Receptor, Parathyroid Hormone, Type 1/genetics
• Renal Veins/drug effects
• Tail
• Transduction, Genetic
• Transgenes

• R01 DK090058 NIDDK NIH HHS
• R21 DK090548 NIDDK NIH HHS
• R21-DK090548 NIDDK NIH HHS
• R01-DK090058 NIDDK NIH HHS

• Ad vector
• Epitope display
• Hypervariable region
• Immune response

• neurodegeneration
• gene therapy
• rna interference
• nanoparticles

• Blood-Brain Barrier/metabolism
• Drug Carriers/chemistry
• Genetic Vectors/metabolism
• Humans
• Nanoparticles/chemistry
• Nervous System Diseases/metabolism
• Nervous System Diseases/pathology
• Nervous System Diseases/therapy
• RNA Interference
• RNA, Small Interfering/administration & dosage
• Translational Research, Biomedical
• Viruses/genetics

The induction of sterilizing T-cell responses to tumors is a major goal in the development of T-cell vaccines for treating cancer. Although specific components of anti-viral CD8+ immunity are well characterized, we still lack the ability to mimic viral CD8+ T-cell responses in therapeutic settings for treating cancers. Infection with the picornavirus Theiler's murine encephalomyelitis virus (TMEV) induces a strong sterilizing CD8+ T-cell response. In the absence of sterilizing immunity, the virus causes a persistent infection. We capitalized on the ability of TMEV to induce strong cellular immunity even under conditions of immune deficiency by modifying the virus to evaluate its potential as a T-cell vaccine. The introduction of defined CD8+ T-cell epitopes into the leader sequence of the TMEV genome generates an attenuated vaccine strain that can efficiently drive CD8+ T-cell responses to the targeted antigen. This virus activates T-cells in a manner that is capable of inducing targeted tissue damage and glucose dysregulation in an adoptive T-cell transfer model of diabetes mellitus. As a therapeutic vaccine for the treatment of established melanoma, epitope-modified TMEV can induce strong cytotoxic T-cell responses and promote infiltration of the T-cells into established tumors, ultimately leading to a delay in tumor growth and improved survival of vaccinated animals. We propose that epitope-modified TMEV is an excellent candidate for further development as a human T-cell vaccine for use in immunotherapy.

• Amino Acid Sequence
• Animals
• Base Sequence
• DNA Primers
• Immunotherapy
• Mice
• Mice, Inbred C57BL
• Molecular Sequence Data
• Polymerase Chain Reaction
• Sequence Homology, Amino Acid
• Theilovirus/immunology
• Viral Vaccines/immunology

• R01 CA104996 NCI NIH HHS
• R01 NS032129 NINDS NIH HHS
• 5R01NS32129 NINDS NIH HHS
• 5R01CA104996 NCI NIH HHS

• Animals
• Chemokine CCL2/administration & dosage
• Ganciclovir/administration & dosage
• Genes, Transgenic, Suicide
• Genetic Therapy/methods
• Genetic Vectors/administration & dosage
• Immunohistochemistry
• Liver Neoplasms, Experimental/pathology
• Liver Neoplasms, Experimental/therapy
• Mice
• Mice, Inbred BALB C
• Neoplasm Metastasis/prevention & control
• Neoplasm Metastasis/therapy
• Th1 Cells/immunology
• Thymidine Kinase/administration & dosage
• Treatment Outcome

Adenovirus (Ad) based gene transfer vectors continue to be the platform of choice for an increasing number of clinical trials worldwide. In fact, within the last five years, the number of clinical trials that utilize Ad based vectors has doubled, indicating growing enthusiasm for the numerous positive characteristics of this gene transfer platform. For example, Ad vectors can be easily and relatively inexpensively produced to high titers in a cGMP compliant manner, can be stably stored and transported, and have a broad applicability for a wide range of clinical conditions, including both gene therapy and vaccine applications. Ad vector based gene transfer will become more useful as strategies to counteract innate and/or pre-existing adaptive immune responses to Ads are developed and confirmed to be efficacious. The approaches attempting to overcome these limitations can be divided into two broad categories: pre-emptive immune modulation of the host, and selective modification of the Ad vector itself. The first category of methods includes the use of immunosuppressive drugs or specific compounds to block important immune pathways, which are known to be induced by Ads. The second category comprises several innovative strategies inclusive of: (1) Ad-capsid-display of specific inhibitors or ligands; (2) covalent modifications of the entire Ad vector capsid moiety; (3) the use of tissue specific promoters and local administration routes; (4) the use of genome modified Ads; and (5) the development of chimeric or alternative serotype Ads. This review article will focus on both the promise and the limitations of each of these immune evasion strategies, and in the process delineate future directions in developing safer and more efficacious Ad-based gene transfer strategies.

• adaptive immunity
• cellular responses
• humoral responses
• innate immunity
• neutralizing antibodies
• recombinant adenovirus

Adenoviral (Ad) vectors have been used for a variety of vaccine applications including cancer and infectious diseases. Traditionally, Ad-based vaccines are designed to express antigens through transgene expression of a given antigen. However, in some cases these conventional Ad-based vaccines have had sub-optimal clinical results. These sub-optimal results are attributed in part to pre-existing Ad serotype 5 (Ad5) immunity. In order to circumvent the need for antigen expression via transgene incorporation, the “antigen capsid-incorporation” strategy has been developed and used for Ad-based vaccine development in the context of a few diseases. This strategy embodies the incorporation of antigenic peptides within the capsid structure of viral vectors. The major capsid protein hexon has been utilized for these capsid incorporation strategies due to hexon's natural role in the generation of anti-Ad immune response and its numerical representation within the Ad virion. Using this strategy, we have developed the means to incorporate heterologous peptide epitopes specifically within the major surface-exposed domains of the Ad capsid protein hexon. Our study herein focuses on generation of multivalent vaccine vectors presenting HIV antigens within the Ad capsid protein hexon, as well as expressing an HIV antigen as a transgene. These novel vectors utilize HVR2 as an incorporation site for a twenty-four amino acid region of the HIV membrane proximal ectodomain region (MPER), derived from HIV glycoprotein gp41 (gp41). Our study herein illustrates that our multivalent anti-HIV vectors elicit a cellular anti-HIV response. Furthermore, vaccinations with these vectors, which present HIV antigens at HVR2, elicit a HIV epitope-specific humoral immune response.

• AIDS Vaccines/genetics
• AIDS Vaccines/immunology
• Adenoviridae/genetics
• Animals
• Blotting, Western
• Cell Line
• Enzyme-Linked Immunosorbent Assay
• Female
• Flow Cytometry
• Genetic Vectors/genetics
• HIV Antibodies/immunology
• HIV Antigens/chemistry
• HIV Antigens/genetics
• HIV Antigens/immunology
• HIV Envelope Protein gp41/chemistry
• HIV Envelope Protein gp41/genetics
• HIV Envelope Protein gp41/immunology
• Humans
• Immunity, Humoral/immunology
• Mice
• Mice, Inbred BALB C
• Peptides/chemical synthesis
• Peptides/chemistry
• Peptides/immunology

• P30 AI027767 NIAID NIH HHS
• 4R33AI076096-03 NIAID NIH HHS
• P30 AR050948 NIAMS NIH HHS
• R33 AI076096 NIAID NIH HHS
• 2P30AI027767 NIAID NIH HHS

• gene therapy
• nanoparticles
• silica
• sirna

• Animals
• Drug Carriers/chemistry
• Gene Silencing/physiology
• Humans
• Materials Testing
• Nanoparticles/chemistry
• Nanoparticles/ultrastructure
• Particle Size
• Porosity
• RNA, Small Interfering/administration & dosage
• RNA, Small Interfering/genetics
• Signal Transduction/physiology
• Silicon Dioxide/chemistry
• Transfection/methods

• R01 CA133697 NCI NIH HHS
• R01 CA133697-02 NCI NIH HHS
• CA 133697 NCI NIH HHS
• CA41996 NCI NIH HHS
• R01 CA041996 NCI NIH HHS

Covalent modification with polyethylene glycol (PEG), a non-toxic polymer used in food, cosmetic and pharmaceutical preparations for over 60 years, can profoundly influence the pharmacokinetic, pharmacologic and toxciologic profile of protein and peptide-based therapeutics. This review summarizes the history of PEGylation and PEG chemistry and highlights the value of this technology in the context of the design and development of recombinant viruses for gene transfer, vaccination and diagnostic purposes. Specific emphasis is placed on the application of this technology to the adenovirus, the most potent viral vector with the most highly characterized toxicity profile to date, in several animal models.

• PEGylation
• adenovirus
• gene therapy
• immune response
• non-viral vectors
• pharmacokinetics
• targeting
• tolerance
• toxicity
• vaccine

The ultimate goal of gene therapy is the replacement of a defective gene sequence with a corrected version to eliminate disease for the lifetime of the patient. This challenging task is not yet accomplished, however significant progress is evident. An initial spate of clinical trials attempting the treatment of haemophilia with gene transfer primarily resulted in the demonstration of good safety profiles, but without efficacy. Subsequent reengineering of vector plasmids and delivery systems resulted in markedly improved outcomes in animal models of the disease. The most recent clinical trial for the treatment of haemophilia B with gene transfer showed transient achievement of efficacy in the highest dose cohort tested, but also exposed a previously hidden barrier to the future success of these treatments. The progress and problems of gene therapies for haemorrhagic disorders will be discussed. This review will concentrate on approaches in or near clinical application.