Construction of adenovirus vectors through Cre-lox recombination

Hardy, S.; Kitamura, Marina; Harris-Stansil, T; Dai, Yifan; Phipps, M L

doi:10.1128/jvi.71.3.1842-1849.1997

Cited by 746 publications

(219 citation statements)

References 26 publications

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“…U62024) at SalI/NotI sites. Subsequently, replication-defective human adenovirus serotype 5 expressing RSV-F, designated Ad5.RSV-F, was generated by loxP homologous recombination and subsequently purified and stored as described previously (6,23,24).…”

Section: Replication-defective Adenovirus Vaccine Preparationmentioning

confidence: 99%

Development of an Adenovirus-Based Respiratory Syncytial Virus Vaccine: Preclinical Evaluation of Efficacy, Immunogenicity, and Enhanced Disease in a Cotton Rat Model

Kim

Okada

Beeler

et al. 2014

J Virol

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The lack of a vaccine against respiratory syncytial virus (RSV) is a challenging and serious gap in preventive medicine. Herein, we characterize the immunogenicity of an adenovirus serotype 5-based RSV vaccine encoding the fusion (F) protein (Ad5.RSV-F) and the protection provided following immunization with Ad5.RSV-F and assess its potential for producing enhanced disease in a cotton rat (CR) model. Animals were immunized intranasally (i.n.) and/or intramuscularly (i.m.) and subsequently challenged with RSV/A/Tracy (i.n.) to assess protection. Robust immune responses were seen in CRs vaccinated with Ad5.RSV-F given i.m. or i.n., and these responses correlated with reduced replication of the virus in noses and lungs after challenge. Neutralizing antibody responses following immunization with a single dose of Ad5.RSV-F at 1 ؋ 10 11 viral particles (v.p.) elicited antibody titers 64-to 256-fold greater than those seen after natural infection. CRs boosted with Ad5.RSV-F i.n. 28 days after an i.m. dose also had significant increases in neutralizing antibody titers. Antibody affinity for different F-protein antigenic sites revealed substantial differences between antibodies elicited by Ad5.RSV-F and those seen after RSV infection; differences in antibody profiles were also seen between CRs given Ad5.RSV-F i.m. and CRs given Ad5.RSV-F i.n. Ad5.RSV-F priming did not result in enhanced disease following live-virus challenge, in contrast to the histopathology seen in CRs given the formalin-inactivated RSV/A/Burnett vaccine. IMPORTANCERespiratory syncytial virus (RSV) is the most common cause of acute lower respiratory infection in infants and young children and a serious health threat in the immunocompromised and the elderly. Infection severity increased in children in an immunization trial, hampering the over 4-decade-long quest for a successful RSV vaccine. In this study, we show that a genetically engineered RSV-F-encoding adenoviral vector provides protective immunity against RSV challenge without enhanced lung disease in cotton rats (CRs). CRs were vaccinated under a number of different regimens, and the immunity induced by the recombinant adenoviral RSV vaccine administered by use of an intramuscular prime-intranasal boost regimen may provide the best protection for young infants and children at risk of RSV infection, since this population is naive to adenoviral preformed immunity. Overall, this report describes a potential RSV vaccine candidate that merits further evaluation in a phase I clinical study in humans.

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Section: Replication-defective Adenovirus Vaccine Preparationmentioning

confidence: 99%

Development of an Adenovirus-Based Respiratory Syncytial Virus Vaccine: Preclinical Evaluation of Efficacy, Immunogenicity, and Enhanced Disease in a Cotton Rat Model

Kim

Okada

Beeler

et al. 2014

J Virol

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“…1A). Subsequently, replication-defective human adenovirus serotype 5, designated as Ad5.MERS-S and Ad5.MERS-S1, were generated by loxP homologous recombination and purified and stored as described previously [26,41,42].…”

Section: Construction Of Recombinant Adenovirus Vectorsmentioning

confidence: 99%

Immunogenicity of an adenoviral-based Middle East Respiratory Syndrome coronavirus vaccine in BALB/c mice

Kim

Okada

Kenniston

et al. 2014

Vaccine

122

145

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A new type of coronavirus has been identified as the causative agent underlying Middle East Respiratory Syndrome (MERS). The MERS coronavirus (MERS-CoV) has spread in the Middle East, but cases originating in the Middle East have also occurred in the European Union and the USA. Eight hundred and thirty-seven cases of MERS-CoV infection have been confirmed to date, including 291 deaths. MERS-CoV has infected dromedary camel populations in the Middle East at high rates, representing an immediate source of human infection. The MERS-CoV spike (S) protein, a characteristic structural component of the viral envelope, is considered as a key target of vaccines against coronavirus infection. In an initial attempt to develop a MERS-CoV vaccine to ultimately target dromedary camels, we constructed two recombinant adenoviral vectors encoding the full-length MERS-CoV S protein (Ad5.MERS-S) and the S1 extracellular domain of S protein (Ad5.MERS-S1). BALB/c mice were immunized with both candidate vaccines intramuscularly and boosted three weeks later intranasally. All the vaccinated animals had antibody responses against spike protein, which neutralized MERS-CoV in vitro. These results show that an adenoviral-based vaccine can induce MERS-CoV-specific immune responses in mice and hold promise for the development of a preventive vaccine that targets the animal reservoir, which might be an effective measure to eliminate transmission of MERS-CoV to humans.

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“…In order to overcome the low efficiency of homologous recombination in mammalian cells, approaches based on bacteriophage P1 Cre/LoxP recombination system have been developed [21][22][23][24][25]. Ad vectors are generated as a result of Cre-mediated site specific recombination between two plasmids after their co-transfection into a suitable cell line expressing Cre recombinase.…”

Section: Ad Vector Construction Strategiesmentioning

confidence: 99%

Production of adenovirus vectors and their use as a delivery system for influenza vaccines

Vemula

Mittal

2010

Expert Opinion on Biological Therapy

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IMPORTANCE OF THE FIELD-With the emergence of highly pathogenic avian influenza H5N1 viruses that have crossed species barriers and are responsible for lethal infections in humans in many countries, there is an urgent need for the development of effective vaccines which can be produced in large quantities at a short notice and confer broad protection against these H5N1 variants. In order to meet the potential global vaccine demand in a pandemic scenario, new vaccine-production strategies must be explored in addition to the currently used egg-based technology for seasonal influenza.AREAS COVERED IN THIS REVIEW-Adenovirus (Ad) based influenza vaccines represent an attractive alternative/supplement to the currently licensed egg-based influenza vaccines. Ad-based vaccines are relatively inexpensive to manufacture, and their production process does not require either chicken eggs or labor intensive and time-consuming processes necessitating enhanced biosafety facilities. Most importantly, in a pandemic situation, this vaccine strategy could offer a stockpiling option to reduce the response time before a strain-matched vaccine could be developed.WHAT THE READER WILL GAIN-This review discusses Ad-vector technology and the current progress in the development of Ad-based influenza vaccines.TAKE HOME MESSAGE-Ad vector-based influenza vaccines for pandemic preparedness are under development to meet the global vaccine demand. Keywords adenoviral vectors; influenza vaccine; pandemic influenza vaccine; vector immunity Highlights• Adenovirus (Ad) vector-based vaccines can be manufactured and stockpiled in large quantities at a short notice inexpensively with cell culture-based technology without the need for enhanced biosafety facilities.• Ad vector-based vaccines induce balanced humoral and cell-mediated immune responses due to activation of innate immunity. • In order to prepare for a potential influenza pandemic, new influenza vaccines should confer broad protection against multiple strains, be produced in a timely manner and be easily stockpiled.• Ad vector-based influenza vaccines have demonstrated considerable potential in preclinical and limited clinical studies.

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Construction of adenovirus vectors through Cre-lox recombination

Cited by 746 publications

References 26 publications

Development of an Adenovirus-Based Respiratory Syncytial Virus Vaccine: Preclinical Evaluation of Efficacy, Immunogenicity, and Enhanced Disease in a Cotton Rat Model

Development of an Adenovirus-Based Respiratory Syncytial Virus Vaccine: Preclinical Evaluation of Efficacy, Immunogenicity, and Enhanced Disease in a Cotton Rat Model

Immunogenicity of an adenoviral-based Middle East Respiratory Syndrome coronavirus vaccine in BALB/c mice

Production of adenovirus vectors and their use as a delivery system for influenza vaccines

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