| Literature DB >> 28211908 |
Kerstin Wernike1, Andrea Aebischer1, Gleyder Roman-Sosa1, Martin Beer1.
Abstract
Schmallenberg virus (SBV) is transmitted by insect vectors, and therefore vaccination is one of the most important tools of disease control. In our study, novel subunit vaccines on the basis of an amino-terminal domain of SBVEntities:
Mesh:
Substances:
Year: 2017 PMID: 28211908 PMCID: PMC5304187 DOI: 10.1038/srep42500
Source DB: PubMed Journal: Sci Rep ISSN: 2045-2322 Impact factor: 4.379
Vaccines and animal groups.
| vaccine | description | dose | animal group |
|---|---|---|---|
| SBV-Gc Amino (DNA) | N terminal 234 residues of SBV Gc in pCI expression vector | 100 μg DNA i.m./mouse | M01 |
| SBV-Gc Amino (protein) | N terminal 234 residues of SBV Gc expressed in HEK-293T cells | 20 μg protein s.c./mouse; 50 μg protein/cattle | M02, C01 |
| SBV-Gc Amino red | reduced form of SBV-Gc Amino (protein) | 20 μg protein s.c./mouse | M03 |
| SBV-Gc Amino E. coli | SBV-Gc Amino expressed in E. coli | 20 μg protein s.c./mouse | M04 |
| SBV-Gn-L-Gc | covalently linked ectodomains of SBV Gn and Gc expressed in HEK-293T cells | 20 μg protein s.c./mouse; 50 μg protein s.c./cattle | M05, C02 |
| AKA-Gc Amino-SBV-Gc Amino | covalently linked SBV-Gc Amino and AKAV-Gc Amino expressed in HEK-293T cells | 20 μg protein s.c./mouse; 50 μg protein s.c./cattle | M06, C03 |
| AKA-Gc Amino | N terminal 234 residues of AKAV Gc expressed in HEK-293T cells | 20 μg protein s.c./mouse | M07 |
| pCI-vector | pCI expression vector | 100 μg pCI MEGA (DNA) i.m./mouse | M08 |
| unvaccinated | — | 100 μl phosphate-buffered saline s.c./mouse | M09, C04 |
Figure 1Body weight after challenge infection of mice with SBV and real-time PCR results of blood samples taken 2 and 7 days after infection.
Figure 2Results of an S-segment based commercial SBV ELISA (depicted by solid lines) and of the micro-neutralization test (shown as bars) of serum samples taken from cattle vaccinated with SBV-Gc Amino, SBV-Gn-L-Gc, AKA-Gc Amino-SBV-Gc Amino, or mock-vaccinated animals.
The time point of challenge infection is indicated in the figure panel showing group C04 (unvaccinated) by a black arrow. The results of individual animals in each group are depicted in different colors; the identical colors are used in the corresponding panels of Figs 3 and 5. The cut-off values of the commercial antibody ELISA are marked by a dotted line.
Figure 3Results of in-house ELISAs of serum samples taken from cattle vaccinated with SBV-Gc Amino, SBV-Gn-L-Gc, AKA-Gc Amino-SBV-Gc Amino, or mock-vaccinated animals.
The time point of challenge infection is indicated in the figure panel showing group C04 (unvaccinated) by a black arrow. The results of individual animals in each group are depicted in different colors; the identical colors are used in the corresponding panels of Figs 2 and 5.
Figure 4Western Blot analysis.
Sera collected at 28 days post challenge from animals of groups C01 and C03 as well as from one control animal from group C04 were analyzed by Western blot using SBV-infected (+) or non-infected cells (−) as antigens. All sera (dilution 1/100) contained Gc-specific antibodies and enabled staining of SBV-Gc (~120 kDa) in infected cell lysates. In contrast, the SBV-N-protein (~25 kDa) was detected only by sera from the infected control animal as well as by one animal from group C01. Staining of Beta Actin was used as a loading control and is shown (cropped) beneath the corresponding serum-staining. Full-length blots are presented in Supplementary Figure 4.
Figure 5Results of the S-segment based real-time PCR of serum samples taken from cattle vaccinated with SBV-Gc Amino, SBV-Gn-L-Gc, AKA-Gc Amino-SBV-Gc Amino, or mock-vaccinated animals.
The results of individual animals in each group are depicted in different colors; the identical colors are used in the corresponding panels of Figs 2 and 3.