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VP2 Exchange and NS3/NS3a Deletion in African Horse Sickness Virus (AHSV) in Development of Disabled Infectious Single Animal Vaccine Candidates for AHSV.

van de Water SG, van Gennip RG, Potgieter CA, Wright IM, van Rijn PA - J. Virol. (2015)

Bottom Line: Single Seg-2 AHSV reassortants showed similar cytopathogenic effects in mammalian cells but displayed different growth kinetics.Reverse genetics for AHSV was also used to study Seg-10 expressing NS3/NS3a proteins.African horse sickness has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates are supposed to be competent vectors.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Central Veterinary Institute of Wageningen UR (CVI), Lelystad, The Netherlands.

No MeSH data available.


Related in: MedlinePlus

Plaque morphology of Seg-10 mutants of AHSV4LP. BSR cell monolayers were infected with Seg-10 mutants of AHSV4LP and grown under overlay medium. At 24, 48, and 72 h postinfection, cells were fixed and immunostained. Separate plaques were semiquantitated as having normal CPE (AHSV4LP), small CPE, and no CPE, corresponding to +, small, and −, respectively, in Table 2.
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Figure 3: Plaque morphology of Seg-10 mutants of AHSV4LP. BSR cell monolayers were infected with Seg-10 mutants of AHSV4LP and grown under overlay medium. At 24, 48, and 72 h postinfection, cells were fixed and immunostained. Separate plaques were semiquantitated as having normal CPE (AHSV4LP), small CPE, and no CPE, corresponding to +, small, and −, respectively, in Table 2.

Mentions: CPE of Seg-10 mutants of AHSV4LP were studied (Table 2). The mutAUG1+2 mutant formed plaques showing normal CPE, whereas mutAUG1+2&STOPS and delLD formed plaques with reduced CPE (small CPE). The delTM1, delTM2, and delLD&delTM1+2 mutants formed plaques without obvious CPE (no CPE). To semiquantitate CPE in more detail, BSR cell monolayers were infected with AHSV4LP, AUG1+2&STOPS, delLD, delTM2, and delLD&delTM1+2 at an appropriate MOI to generate foci of infected cells that were investigated by IPMAs (Fig. 3). The mutAUG1+2&STOPS and delLD mutants induced small CPE. This smaller CPE seems to be associated with a delay of CPE induction of ∼1 day. CPE of AHSV4LP at 24 hpi and 48 hpi were comparable to those of AUG1+2&STOPS and delLD at 48 hpi and 72 hpi, respectively (Fig. 3). The delTM2 and delLD&delTM1+2 mutants induced no CPE up to 72 hpi. Apparently, CPE is associated with NS3 expression and in particular with the expression of one or two transmembrane regions.


VP2 Exchange and NS3/NS3a Deletion in African Horse Sickness Virus (AHSV) in Development of Disabled Infectious Single Animal Vaccine Candidates for AHSV.

van de Water SG, van Gennip RG, Potgieter CA, Wright IM, van Rijn PA - J. Virol. (2015)

Plaque morphology of Seg-10 mutants of AHSV4LP. BSR cell monolayers were infected with Seg-10 mutants of AHSV4LP and grown under overlay medium. At 24, 48, and 72 h postinfection, cells were fixed and immunostained. Separate plaques were semiquantitated as having normal CPE (AHSV4LP), small CPE, and no CPE, corresponding to +, small, and −, respectively, in Table 2.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4524073&req=5

Figure 3: Plaque morphology of Seg-10 mutants of AHSV4LP. BSR cell monolayers were infected with Seg-10 mutants of AHSV4LP and grown under overlay medium. At 24, 48, and 72 h postinfection, cells were fixed and immunostained. Separate plaques were semiquantitated as having normal CPE (AHSV4LP), small CPE, and no CPE, corresponding to +, small, and −, respectively, in Table 2.
Mentions: CPE of Seg-10 mutants of AHSV4LP were studied (Table 2). The mutAUG1+2 mutant formed plaques showing normal CPE, whereas mutAUG1+2&STOPS and delLD formed plaques with reduced CPE (small CPE). The delTM1, delTM2, and delLD&delTM1+2 mutants formed plaques without obvious CPE (no CPE). To semiquantitate CPE in more detail, BSR cell monolayers were infected with AHSV4LP, AUG1+2&STOPS, delLD, delTM2, and delLD&delTM1+2 at an appropriate MOI to generate foci of infected cells that were investigated by IPMAs (Fig. 3). The mutAUG1+2&STOPS and delLD mutants induced small CPE. This smaller CPE seems to be associated with a delay of CPE induction of ∼1 day. CPE of AHSV4LP at 24 hpi and 48 hpi were comparable to those of AUG1+2&STOPS and delLD at 48 hpi and 72 hpi, respectively (Fig. 3). The delTM2 and delLD&delTM1+2 mutants induced no CPE up to 72 hpi. Apparently, CPE is associated with NS3 expression and in particular with the expression of one or two transmembrane regions.

Bottom Line: Single Seg-2 AHSV reassortants showed similar cytopathogenic effects in mammalian cells but displayed different growth kinetics.Reverse genetics for AHSV was also used to study Seg-10 expressing NS3/NS3a proteins.African horse sickness has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates are supposed to be competent vectors.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Central Veterinary Institute of Wageningen UR (CVI), Lelystad, The Netherlands.

No MeSH data available.


Related in: MedlinePlus