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Evaluation of Different DNA Vaccines against Porcine Reproductive and Respiratory Syndrome (PRRS) in Pigs.

Petrini S, Ramadori G, Villa R, Borghetti P, de Angelis E, Cantoni AM, Corradi A, Amici A, Ferrari M - Vaccines (Basel) (2013)

Bottom Line: Only vaccines A and B were able to reduce the clinical signs of the infection.Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups.In the other groups, the IFN-g were detected after challenge infection.

View Article: PubMed Central - PubMed

Affiliation: Umbria and Marche Experimental Zooprophylaxis Institute, via Gaetano Salvemini 1, Perugia 06126, Italy. s.petrini@izsum.it.

ABSTRACT
In veterinary medicine, there have been different experiences with the plasmid DNA vaccination. In this area and with the hypothesis to demonstrate the effectiveness of different plasmids encoding porcine respiratory and reproductive syndrome (PRRS), five DNA vaccines against PRRS were evaluated for their innocuity and efficacy in pigs. Eighteen animals were divided into five groups which were injected with five (A, B, C, D, E) different DNA vaccines. Albeit, none of the proposed vaccines were able to protect the animals against PRRS virus. Only vaccines A and B were able to reduce the clinical signs of the infection. ELISA IgM were detected 30 days after the first vaccination in the pigs injected by Vaccine A or B. ELISA IgG were detected 90 days after the first vaccination in the pigs injected by Vaccine B or C. Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups. In the pigs inoculated with Vaccine C, IFN-g were detected 90 days after first vaccination, and after challenge exposure they increased. In the other groups, the IFN-g were detected after challenge infection. Pigs injected with each of the vaccines A, B, C, D and E showed a significantly higher level of CD4(-)CD8⁺ lymphocytes (p < 0.001) after infection in comparison with their controls.

No MeSH data available.


Related in: MedlinePlus

Plasmid pVAX1-48CpG-NeuL-ORF5.
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vaccines-01-00463-f003: Plasmid pVAX1-48CpG-NeuL-ORF5.

Mentions: All plasmids derived from pVAX1 (Invitrogen, San Diego, CA, USA). Plasmids were constructed by cloning PRRS genes encoding GP4 and GP5 into different plasmids: pVAX1-48CpG-NeuL-ORF4 (Figure 2); pVAX1-48CpG-NeuL-ORF5 (Figure 3); pVAX1-48CpG-UbilacI-ORF4 (Figure 4); pVAX1-48CpG-UbilacI-ORF5 (Figure 5). NeuL sequence was cloned into EcoRI end Hind III restriction sites and encoded for strong proteic secretion signal. Therefore, NeuL should allow for the protein processing by the endoplasmatic reticulum that is a step required for a new synthesized protein to be trans-located to cellular membranes and/or be secreted. These events allow for the viral antigen produced by the plasmid to be presented to the immune system in order to stimulate an antibody response. The sequence neuL includes Her-2/neu 5'UTR. The secretion signal DNA fragment was obtained by enzymatic amplification of DNA using the pCMV-ECD-TM vector [9,10] as a template, T7 primer (5'-TAATACGACTCACTATATAGGG-3') as a sense oligonucleotide and an oligonucleotide antisense having a terminal EcoR I site, “neuL” antisense EcoR I (5'-CATGGAATTCCGCGATTCCGGGGGGCAGGA-3'). The sequence UbiLacI encodes for a signal that leads to the proteasomal degradation [11]. This sequence was generated by polymerase chain reaction (PCR) from reference sequence [12] and cloned into Hind III and EcoR I restriction sites in line with viral sequence using the following primers:


Evaluation of Different DNA Vaccines against Porcine Reproductive and Respiratory Syndrome (PRRS) in Pigs.

Petrini S, Ramadori G, Villa R, Borghetti P, de Angelis E, Cantoni AM, Corradi A, Amici A, Ferrari M - Vaccines (Basel) (2013)

Plasmid pVAX1-48CpG-NeuL-ORF5.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494207&req=5

vaccines-01-00463-f003: Plasmid pVAX1-48CpG-NeuL-ORF5.
Mentions: All plasmids derived from pVAX1 (Invitrogen, San Diego, CA, USA). Plasmids were constructed by cloning PRRS genes encoding GP4 and GP5 into different plasmids: pVAX1-48CpG-NeuL-ORF4 (Figure 2); pVAX1-48CpG-NeuL-ORF5 (Figure 3); pVAX1-48CpG-UbilacI-ORF4 (Figure 4); pVAX1-48CpG-UbilacI-ORF5 (Figure 5). NeuL sequence was cloned into EcoRI end Hind III restriction sites and encoded for strong proteic secretion signal. Therefore, NeuL should allow for the protein processing by the endoplasmatic reticulum that is a step required for a new synthesized protein to be trans-located to cellular membranes and/or be secreted. These events allow for the viral antigen produced by the plasmid to be presented to the immune system in order to stimulate an antibody response. The sequence neuL includes Her-2/neu 5'UTR. The secretion signal DNA fragment was obtained by enzymatic amplification of DNA using the pCMV-ECD-TM vector [9,10] as a template, T7 primer (5'-TAATACGACTCACTATATAGGG-3') as a sense oligonucleotide and an oligonucleotide antisense having a terminal EcoR I site, “neuL” antisense EcoR I (5'-CATGGAATTCCGCGATTCCGGGGGGCAGGA-3'). The sequence UbiLacI encodes for a signal that leads to the proteasomal degradation [11]. This sequence was generated by polymerase chain reaction (PCR) from reference sequence [12] and cloned into Hind III and EcoR I restriction sites in line with viral sequence using the following primers:

Bottom Line: Only vaccines A and B were able to reduce the clinical signs of the infection.Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups.In the other groups, the IFN-g were detected after challenge infection.

View Article: PubMed Central - PubMed

Affiliation: Umbria and Marche Experimental Zooprophylaxis Institute, via Gaetano Salvemini 1, Perugia 06126, Italy. s.petrini@izsum.it.

ABSTRACT
In veterinary medicine, there have been different experiences with the plasmid DNA vaccination. In this area and with the hypothesis to demonstrate the effectiveness of different plasmids encoding porcine respiratory and reproductive syndrome (PRRS), five DNA vaccines against PRRS were evaluated for their innocuity and efficacy in pigs. Eighteen animals were divided into five groups which were injected with five (A, B, C, D, E) different DNA vaccines. Albeit, none of the proposed vaccines were able to protect the animals against PRRS virus. Only vaccines A and B were able to reduce the clinical signs of the infection. ELISA IgM were detected 30 days after the first vaccination in the pigs injected by Vaccine A or B. ELISA IgG were detected 90 days after the first vaccination in the pigs injected by Vaccine B or C. Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups. In the pigs inoculated with Vaccine C, IFN-g were detected 90 days after first vaccination, and after challenge exposure they increased. In the other groups, the IFN-g were detected after challenge infection. Pigs injected with each of the vaccines A, B, C, D and E showed a significantly higher level of CD4(-)CD8⁺ lymphocytes (p < 0.001) after infection in comparison with their controls.

No MeSH data available.


Related in: MedlinePlus