High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells.
This work investigated the replication kinetics of the four dengue virus serotypes (DEN-1 to DEN-4), including dengue virus type 4 (DEN-4) recovered from an infectious cDNA clone, in Vero cells and in MRC-5 cells grown on Cytodex 1 microcarriers. DEN-1 strain Hawaii, DEN-2 strain NGC, DEN-3 strain H...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2008-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2265545?pdf=render |
| _version_ | 1811268612834459648 |
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| author | Chia-Chyi Liu Shiang-Chi Lee Michael Butler Suh-Chin Wu |
| author_facet | Chia-Chyi Liu Shiang-Chi Lee Michael Butler Suh-Chin Wu |
| author_sort | Chia-Chyi Liu |
| collection | DOAJ |
| description | This work investigated the replication kinetics of the four dengue virus serotypes (DEN-1 to DEN-4), including dengue virus type 4 (DEN-4) recovered from an infectious cDNA clone, in Vero cells and in MRC-5 cells grown on Cytodex 1 microcarriers. DEN-1 strain Hawaii, DEN-2 strain NGC, DEN-3 strain H-87, and DEN-4 strain H-241 , and DEN-4 strain 814669 derived from cloned DNA, were used to infect Vero cells and MRC-5 cells grown in serum-free or serum-containing microcarrier cultures. Serum-free and serum-containing cultures were found to yield comparable titers of these viruses. The cloned DNA-derived DEN-4 started genetically more homogeneous was used to investigate the genetic stability of the virus propagated in Vero cells and MRC-5 cells. Sequence analysis revealed that the DEN-4 propagated in MRC-5 cells maintained a high genetic stability, compared to the virus propagated in Vero cells. Amino acid substitutions of Gly(104)Cys and Phe(108)Ile were detected at 70%, 60%, respectively, in the envelope (E) protein of DEN-4 propagated in Vero cells, whereas a single mutation of Glu(345)Lys was detected at 50% in E of the virus propagated in MRC-5 cells. Sequencing of multiple clones of three separate DNA fragments spanning 40% of the genome also indicated that DEN-4 propagated in Vero cells contained a higher number of mutations than the virus growing in MRC-5 cells. Although Vero cells yielded a peak virus titer approximately 1 to 17 folds higher than MRC-5 cells, cloned DEN-4 from MRC-5 cells maintained a greater stability than the virus from Vero cells. Serum-free microcarrier cultures of MRC-5 cells offer a potentially valuable system for the large-scale production of live-attenuated DEN vaccines. |
| first_indexed | 2024-04-12T21:26:05Z |
| format | Article |
| id | doaj.art-ae06946eb62547abaf4fa4730624459c |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T21:26:05Z |
| publishDate | 2008-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ae06946eb62547abaf4fa4730624459c2022-12-22T03:16:11ZengPublic Library of Science (PLoS)PLoS ONE1932-62032008-01-0133e181010.1371/journal.pone.0001810High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells.Chia-Chyi LiuShiang-Chi LeeMichael ButlerSuh-Chin WuThis work investigated the replication kinetics of the four dengue virus serotypes (DEN-1 to DEN-4), including dengue virus type 4 (DEN-4) recovered from an infectious cDNA clone, in Vero cells and in MRC-5 cells grown on Cytodex 1 microcarriers. DEN-1 strain Hawaii, DEN-2 strain NGC, DEN-3 strain H-87, and DEN-4 strain H-241 , and DEN-4 strain 814669 derived from cloned DNA, were used to infect Vero cells and MRC-5 cells grown in serum-free or serum-containing microcarrier cultures. Serum-free and serum-containing cultures were found to yield comparable titers of these viruses. The cloned DNA-derived DEN-4 started genetically more homogeneous was used to investigate the genetic stability of the virus propagated in Vero cells and MRC-5 cells. Sequence analysis revealed that the DEN-4 propagated in MRC-5 cells maintained a high genetic stability, compared to the virus propagated in Vero cells. Amino acid substitutions of Gly(104)Cys and Phe(108)Ile were detected at 70%, 60%, respectively, in the envelope (E) protein of DEN-4 propagated in Vero cells, whereas a single mutation of Glu(345)Lys was detected at 50% in E of the virus propagated in MRC-5 cells. Sequencing of multiple clones of three separate DNA fragments spanning 40% of the genome also indicated that DEN-4 propagated in Vero cells contained a higher number of mutations than the virus growing in MRC-5 cells. Although Vero cells yielded a peak virus titer approximately 1 to 17 folds higher than MRC-5 cells, cloned DEN-4 from MRC-5 cells maintained a greater stability than the virus from Vero cells. Serum-free microcarrier cultures of MRC-5 cells offer a potentially valuable system for the large-scale production of live-attenuated DEN vaccines.http://europepmc.org/articles/PMC2265545?pdf=render |
| spellingShingle | Chia-Chyi Liu Shiang-Chi Lee Michael Butler Suh-Chin Wu High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells. PLoS ONE |
| title | High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells. |
| title_full | High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells. |
| title_fullStr | High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells. |
| title_full_unstemmed | High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells. |
| title_short | High genetic stability of dengue virus propagated in MRC-5 cells as compared to the virus propagated in vero cells. |
| title_sort | high genetic stability of dengue virus propagated in mrc 5 cells as compared to the virus propagated in vero cells |
| url | http://europepmc.org/articles/PMC2265545?pdf=render |
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