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Cloning and identification of PK15 cells for enhanced replication of classical swine fever virus

Mei Yin
Mei Yin
, 
Dongfang Hu
Dongfang Hu
, 
Peng Li
Peng Li
, 
Lingyun Kong
Lingyun Kong
, 
Hongmei Ning
Hongmei Ning
, 
Feng Yue
Feng Yue
, 
Jinqing Jiang
Jinqing Jiang
 oraz 
Xuannian Wang
Xuannian Wang
   | 24 mar 2020
Journal of Veterinary Research's Cover Image
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Data publikacji: 24 mar 2020
Zakres stron: 9 - 14
Otrzymano: 17 lip 2019
Przyjęty: 27 lut 2020
DOI: https://doi.org/10.2478/jvetres-2020-0020
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© 2020 M. Yin et al. published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

The percentages of CSFV-infected cells in the parent cell line and cloned PK15 cells. A – mock-infected PK15 cells (negative control); B – high-permissive (clone 1A6); C – low-permissive (clone 3B1); D – infected parent PK15 cells; E – percentages of positively stained cells. The cells were infected with CSFV and stained with CSFV-specific monoclonal antibodies (400×). Five visual fields were randomly selected to be photographed in each group. The percentages of positively stained cells in the photos were counted and shown as mean ± SD. * P < 0.05, ** P < 0.01
The percentages of CSFV-infected cells in the parent cell line and cloned PK15 cells. A – mock-infected PK15 cells (negative control); B – high-permissive (clone 1A6); C – low-permissive (clone 3B1); D – infected parent PK15 cells; E – percentages of positively stained cells. The cells were infected with CSFV and stained with CSFV-specific monoclonal antibodies (400×). Five visual fields were randomly selected to be photographed in each group. The percentages of positively stained cells in the photos were counted and shown as mean ± SD. * P < 0.05, ** P < 0.01

Fig. 2

Growth curves of parent PK15, clone PK15-1A6, and PK15-3B1 cells. The cells were trypsinised and counted at 8, 12, 24, 36, and 48 h post seeding. The data represent the mean ± SD of three independent experiments performed. The differences between the number of parent cells and that of PK15-1A6 cells are shown above the squares. The differences between the number of parent cells and that of PK15-3B1 cells are shown below the triangles. * P < 0.05, ** P < 0.01
Growth curves of parent PK15, clone PK15-1A6, and PK15-3B1 cells. The cells were trypsinised and counted at 8, 12, 24, 36, and 48 h post seeding. The data represent the mean ± SD of three independent experiments performed. The differences between the number of parent cells and that of PK15-1A6 cells are shown above the squares. The differences between the number of parent cells and that of PK15-3B1 cells are shown below the triangles. * P < 0.05, ** P < 0.01

Fig. 3

Mean CSFV titres produced by the parent PK15, PK15-1A6, and PK15-3B1 cells. The replication abilities of CSFV in each type of PK15 cells (TCID50) were detected by the IPMA method. ** P < 0.01
Mean CSFV titres produced by the parent PK15, PK15-1A6, and PK15-3B1 cells. The replication abilities of CSFV in each type of PK15 cells (TCID50) were detected by the IPMA method. ** P < 0.01

Fig. 4

IPMA results demonstrating the replication abilities of CSFV in high- and low-permissive PK15 cells at different time points. Different cell lines were fixed at 6, 12, 18, 24, 36, and 48 h post CSFV infection and stained with CSFV-specific antibodies followed by HRP-conjugated secondary antibodies (400×)
IPMA results demonstrating the replication abilities of CSFV in high- and low-permissive PK15 cells at different time points. Different cell lines were fixed at 6, 12, 18, 24, 36, and 48 h post CSFV infection and stained with CSFV-specific antibodies followed by HRP-conjugated secondary antibodies (400×)

Fig. 5

The synthesis of CSFV genome in parent PK15 cells and PK15-1A6 (HP) and PK15-3B1 (LP) cell lines. The genomic RNA extractions from infected cells were quantitated by RT-PCR (amplification of partial E2 gene). The copy numbers of CSFV were determined by referring to a standard curve. ** P < 0.01
The synthesis of CSFV genome in parent PK15 cells and PK15-1A6 (HP) and PK15-3B1 (LP) cell lines. The genomic RNA extractions from infected cells were quantitated by RT-PCR (amplification of partial E2 gene). The copy numbers of CSFV were determined by referring to a standard curve. ** P < 0.01
eISSN:
2450-8608
Język:
Angielski
Częstotliwość wydawania:
4 razy w roku
Dziedziny czasopisma:
Life Sciences, Molecular Biology, Microbiology and Virology, other, Medicine, Veterinary Medicine
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