Construction and characterization of a recombinant human adenovirus type 3 vector containing two foreign neutralizing epitopes in hexon

Xue, Chunyan; Tian, Xingui; Li, Xiao; Zhou, Zhichao; Su, Xiaobo; Zhou, Rong

doi:10.1016/j.virusres.2014.01.027

Cited by 8 publications

(8 citation statements)

References 26 publications

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“…Generation time for the recombinant adenoviruses was recorded as >12 h while for dAd5 it was ≥12 h. The delayed appearance of progeny viruses in the cells infected with recombinant hAd5 viruses may have been due to increased size of the genome compared to dAd5. The growth kinetics of the viruses was found to be similar to those of the previous study involving hAd5 [ 39 ] and hAd3 [ 40 ]. Maximum growth was recorded during first 48 h which decreased later on which may be due to unavailability of sufficient healthy cells for a further round of virus replication.…”

Section: Discussionsupporting

confidence: 79%

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Construction and characterization of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus capsid proteins of Indian vaccine strain, O/IND/R2/75

Kumar¹,

Sreenivasa²,

Tamilselvan³

2015

Vet World

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Aim:Generation of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus (FMDV) capsid protein genes along with full-length 2B, 3B and 3Cpro and its characterization.Materials and Methods:FMD viral RNA isolation, cDNA synthesis, and polymerase chain reaction were performed to synthesize expression cassettes (P1-2AB3BCwt and P1-2AB3BCm) followed by cloning in pShuttle-CMV vector. Chemically competent BJ5183-AD-1 cells were transformed with the recombinant pShuttle-CMV to produce recombinant adenoviral plasmids. HEK-293 cells were transfected with the recombinant adenoviral plasmids to generate recombinant adenoviruses (hAd5/P1-2AB3BCwt and hAd5/P1-2AB3BCm). Expression of the target proteins was analyzed by sandwich ELISA and indirect immunofluorescence assay. The recombinant adenoviruses were purified and concentrated by CsCl density gradient ultracentrifugation. Growth kinetics and thermostability of the recombinant adenoviruses were compared with that of non-recombinant replication-defective adenovirus (dAd5).Results:The recombinant adenoviruses containing capsid protein genes of the FMDV O/IND/R2/75 were generated and amplified in HEK-293 cells. The titer of the recombinant adenoviruses was approximately 108, 109.5 and 1011 TCID50/ml in supernatant media, cell lysate and CsCl purified preparation, respectively. Expression of the FMDV capsid protein was detectable in sandwich ELISA and confirmed by immunofluorescence assay. Growth kinetics of the recombinant adenoviruses did not reveal a significant difference when compared with that of dAd5. A decrement of up to 10-fold at 4°C and 21-fold at 37°C was recorded in the virus titers during 60 h incubation period and found to be statistically significant (p<0.01).Conclusion:Recombinant adenoviruses expressing capsid proteins of the FMDV O/IND/R2/75 were constructed and produced in high titers. In vitro expression of the target proteins in the adenovirus vector system was detected by sandwich ELISA and immunofluorescence assay.

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Section: Discussionsupporting

confidence: 79%

“…In a study, Xue et al . [ 40 ] demonstrated that thermostability of the hAd5 at 45°C decrease more rapidly.…”

Section: Discussionmentioning

confidence: 99%

Construction and characterization of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus capsid proteins of Indian vaccine strain, O/IND/R2/75

Kumar¹,

Sreenivasa²,

Tamilselvan³

2015

Vet World

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“…This approach most commonly targets linear epitopes and can be used for immunotherapeutic and immunoprophylactic purposes. [1][2][3] Numerous gene delivery systems exist serving different purposes. 4 Both non-viral and viral delivery systems can be used for epitope-based vaccination.…”

Section: Introductionmentioning

confidence: 99%

Live attenuated influenza vaccine viral vector induces functional cytotoxic T-cell immune response against foreign CD8+ T-cell epitopes inserted into NA and NS1 genes using the 2A self-cleavage site

Kotomina

Korenkov

Matyushenko

et al. 2018

Human Vaccines & Immunotherapeutics

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The development of viral vector vaccines against various pathogens for which conventional vaccination approaches are not applicable has been a priority for a number of years. One promising approach is the insertion of immunodominant conservative cytotoxic T-cell (CTL) epitopes into the genome of a viral vector, which then delivers these epitopes to target cells, inducing immunity. Many different viruses have been assessed as viral vectors for CTL-based vaccines, but only a few of them are clinically relevant, mainly because of safety issues and limited knowledge about their performance in humans. In this regard, the use of licensed cold-adapted live attenuated influenza vaccine (LAIV) viruses as a vector delivery system has clear advantages for CTL-based vector vaccines against other respiratory pathogens: LAIV is known to induce all arms of the adaptive immune system and is administered via nasal spray, and its production process is relatively easy and inexpensive. Here we present the first results of the use of an LAIV backbone for designing a CTL epitope-based vaccine against respiratory syncytial virus (RSV). The chimeric LAIV-RSV vaccine candidates were attenuated in mice and induced strong, fully functional CTL immunity in this animal model.

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“…Due to variations in the HVR loop aa sizes and possibly surface-exposure among AdV types (e.g., HAdV5 HVR1 has 30 aa vs. HAdV48 HVR1 which has 8 aa), it is important to determine whether alternative AdV types can be successfully used to display antigens via different HVRs. Epitope display via hexon using alternative AdV types was demonstrated with HAdV3 [ 64 , 65 ], and simian adenovirus 25 (SAdV25) [ 66 , 67 ] vectors, with both vectors inducing potent immune responses in mice. For the HAdV3 vector, a 15 aa enterovirus epitope was successfully displayed via HVR1, HVR2 and HVR7, whereas display via HVR4 and HVR5 was unsuccessful [ 64 , 65 ].…”

Section: Hexon Antigen Displaymentioning

confidence: 99%

Progress in Adenoviral Capsid-Display Vaccines

Vujadinovic

Vellinga

2018

Biomedicines

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Adenoviral vectored vaccines against infectious diseases are currently in clinical trials due to their capacity to induce potent antigen-specific B- and T-cell immune responses. Heterologous prime-boost vaccination with adenoviral vector and, for example, adjuvanted protein-based vaccines can further enhance antigen-specific immune responses. Although leading to potent immune responses, these heterologous prime-boost regimens may be complex and impact manufacturing costs limiting efficient implementation. Typically, adenoviral vectors are engineered to genetically encode a transgene in the E1 region and utilize the host cell machinery to express the encoded antigen and thereby induce immune responses. Similarly, adenoviral vectors can be engineered to display foreign immunogenic peptides on the capsid-surface by insertion of antigens in capsid proteins hexon, fiber and protein IX. The ability to use adenoviral vectors as antigen-display particles, with or without using the genetic vaccine function, greatly increases the versatility of the adenoviral vector for vaccine development. This review describes the application of adenoviral capsid antigen-display vaccine vectors by focusing on their distinct advantages and possible limitations in vaccine development.

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Construction and characterization of a recombinant human adenovirus type 3 vector containing two foreign neutralizing epitopes in hexon

Cited by 8 publications

References 26 publications

Construction and characterization of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus capsid proteins of Indian vaccine strain, O/IND/R2/75

Construction and characterization of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus capsid proteins of Indian vaccine strain, O/IND/R2/75

Live attenuated influenza vaccine viral vector induces functional cytotoxic T-cell immune response against foreign CD8+ T-cell epitopes inserted into NA and NS1 genes using the 2A self-cleavage site

Progress in Adenoviral Capsid-Display Vaccines

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