Adenoviridae ; Adenovirus Vaccines ; China ; Humans ; Vaccination ; Vaccines

The avirulent QU strain of fowl adenovirus, a member of duck adenovirus type 1, could be a potential vector in recombinant vaccine development. To identify a non-essential region for replication of QU virus, a 3.4 kb fragment near the E4 region of QU virus genome was amplified by PCR to construct a plasmid pADGFP, in which ORF1, ORF8 and ORF9 was replaced with a system expressing enhanced green fluorescence protein. Further, a recombinant virus rQUGFP was constructed by homologous recombination after pADGFP and QU virus were co-transfected into chick embryo fibroblast. The one step growth curve of the rQUGFP was found to be identical with that of parent QU virus and the TCID50 titers of different generation recombinants maintained stable. These findings suggest that the region including ORF1, ORF8 and ORF9 of QU virus genome is dispensable for virus replication, and the foreign gene inserted into virus genome can be efficiently and stably expressed. The work lays the foundation for further studies of developing this virus as a vector of recombinant vaccine.
Adenovirus E4 Proteins ; genetics ; immunology ; Animals ; Fowl adenovirus A ; classification ; genetics ; Genes, Viral ; genetics ; Genetic Vectors ; genetics ; Open Reading Frames ; genetics ; Recombination, Genetic ; Transfection ; Vaccines, Synthetic ; biosynthesis ; genetics ; immunology ; Viral Vaccines ; biosynthesis ; genetics ; immunology ; Virus Replication

OBJECTIVETo complete the full-length sequencing of the human adenovirus type 7 vaccine strain (Ad7v) for novel vector constructing.

METHODSThe Ad7v DNA was digested with SalI and the 17.5-68.0 map unit (mu) fragment was cloned and sequenced. The homology of encoding sequence of Ad7v hexon to those of group A,C,D,E,F and other numbers of group B was accomplished with the software CLUSTAL.V. The three-dimensional structure of the Ad7v hexon was predicted with the RasMo12.71.

RESULTSThe fragment contains 17,596 bp, part of E2 and late gene L1, L2 and L3 were encoded by this region. Polypeptide encoded by hexon gene lies in L3 region, which is composed of 934 amino acids. Multiple sequence alignment with the other nine known hexon protein sequences suggested that the variable sequences are mainly concentrated on seven regions, namely hypervariable regions (HVRs). The seven HVRs are related to type-specificity and group-specificity. The three-dimensional structure of the Ad7v hexon revealed that the variable regions are located in the I1 and I2 loops of the molecule mostly on the tower of the hexon.

CONCLUSIONThe full-length genome sequencing of Ad7v was accomplished at last. Since the deduced amino acid sequence of Ad7v hexon was quite different from other adenoviral vectors such as Ad5 and Ad2, this virus can be potentially used for the construction of novel gene delivery vectors to counterpart the immunity to the vectors widely used at present.

Adenovirus E3 Proteins ; chemistry ; Adenoviruses, Human ; genetics ; Amino Acid Sequence ; Base Sequence ; Capsid Proteins ; chemistry ; Genetic Vectors ; Sequence Homology, Amino Acid ; Viral Vaccines ; chemistry

The major immunogenic protein capsid (Cap) of porcine circovirus type 2 (PCV2) is critical to induce neutralizing antibodies and protective immune response against PCV2 infection. This study was conducted to investigate the immune response of recombinant adenovirus expressing PCV2b Cap and C-terminal domain of Yersinia pseudotuberculosis invasin (Cap-InvC) fusion protein in pigs. The recombinant adenovirus rAd-Cap-InvC, rAd-Cap and rAd were generated and used to immunize pigs. The phosphate-buffered saline was used as negative control. The specific antibodies levels in rAd-Cap-InvC and ZJ/C-strain vaccine groups were higher than that of rAd-Cap group (p < 0.05), and the neutralization antibody titer in rAd-Cap-InvC group was significantly higher than those of other groups during 21–42 days post-immunization (DPI). Moreover, lymphocyte proliferative level, interferon-γ and interleukin-13 levels in rAd-Cap-InvC group were increased compared to rAd-Cap group (p < 0.05). After virulent challenge, viruses were not detected from the blood samples in rAd-Cap-InvC and ZJ/C-strain vaccine groups after 49 DPI. And the respiratory symptom, rectal temperature, lung lesion and lymph node lesion were minimal and similar in the ZJ/C-strain and rAd-Cap-InVC groups. In conclusion, our results demonstrated that rAd-Cap-InvC was more efficiently to stimulate the production of antibody and protect pigs from PCV2 infection. We inferred that InvC is a good candidate gene for further development and application of PCV2 genetic engineering vaccine.
Adenoviridae ; Adenovirus Vaccines ; Antibodies ; Antibodies, Neutralizing ; Capsid ; Capsid Proteins ; Circovirus ; Genetic Engineering ; Immunization ; Interleukin-13 ; Lung ; Lymph Nodes ; Lymphocytes ; Swine ; Yersinia pseudotuberculosis

OBJECTIVETo construct and characterize recombinant adenoviruses containing glycoprotein (GP) gene from rabies virus CVS-N2C strain.

METHODSTo obtain the recombinant adenovirus by pAdEasy system, identify recombinant virus with cDNA sequencing, Northern blot, Dot blot, Western blot and challenge-protection experiment of mice.

RESULTSRecombinant adenovirus showed typical adenovirus morphological characteristics; the viral genome was stable; GP specific mRNA and presence of glycoprotein were determined in rAdGPcvs and rAdGPcvs' infected cells. The glycoprotein produced by recombinant adenovirus had a molecular mass of 66,000, which was similar to that of natural glycoprotein. In the group of rAdGPcvs immunized mice, 87.5%-100% of mice survived from a 35.8LD50/38.0LD50 lethal rabies intracerebral challenge. Finally 73.3%-83.3% of the mice that had received eAdGPcvs survived, and all the Ad5 immunized mice succumbed to rabies.

CONCLUSIONRecombinant adenovirus rAdGPcvs and rAdGPcvs' hold great potential to be developed as recombinant rabies vaccines, and at the same time, it is actually the first study that on high neuropathogenicity rabies CVS-N2C glycoprotein based adenoviral recombinants.

Adenovirus E3 Proteins ; biosynthesis ; genetics ; immunology ; Adenoviruses, Human ; genetics ; Animals ; Antibodies, Viral ; biosynthesis ; genetics ; immunology ; Antigens, Viral ; Base Sequence ; Genetic Vectors ; Glycoproteins ; biosynthesis ; genetics ; immunology ; Humans ; Mice ; Molecular Sequence Data ; Nucleoproteins ; biosynthesis ; genetics ; immunology ; Rabies ; immunology ; prevention & control ; Rabies Vaccines ; biosynthesis ; genetics ; immunology ; Rabies virus ; genetics ; immunology ; Vaccines, Synthetic ; biosynthesis ; genetics ; immunology ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology ; Virus Replication

We have previously evaluated a Semliki Forest virus (SFV) replicon vectored DNA vaccine (pSFV1CS2-E2) and a recombinant adenovirus (rAdV-E2) expressing the E2 glycoprotein of classical swine fever virus (CSFV) in pigs. The results showed that the immunized pigs were protected from virulent challenge, but few pigs showed short-term fever and occasional pathological changes following virulent challenge. To enhance the immunogenecity of the vaccines, we tried a prime-boost vaccination strategy using a combination of prime with pSFV1CS2-E2 followed by boost with rAdV-E2. The results showed that all the immunized pigs developed high-level CSFV-specific antibodies following prime-boost immunization. When challenged with virulent CSFV, the immunized pigs (n = 5) from the heterologous boost group showed no clinical symptoms, and CSFV RNA was not detected following challenge, whereas one of five pigs from the homologous boost group developed short-term fever and CSFV RNA was detected. This demonstrates that the heterologous prime-boost vaccination regime has the potential to prevent against virulent challenge.
Adenoviridae ; genetics ; metabolism ; Adenovirus E2 Proteins ; genetics ; immunology ; Animals ; Classical Swine Fever ; immunology ; prevention & control ; Classical swine fever virus ; genetics ; immunology ; Genetic Vectors ; Immunization, Secondary ; Replicon ; genetics ; Semliki forest virus ; genetics ; metabolism ; Swine ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; genetics ; metabolism ; Viral Vaccines ; immunology