| Autor: |
Carter MH; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA., Gribble J; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA., Diller JR; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA., Denison MR; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA.; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA., Mirza SA; Centers for Disease Control and Prevention, Atlanta, GA 30329, USA., Chappell JD; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA., Halasa NB; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA., Ogden KM; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA.; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA. |
| Abstrakt: |
Human rotaviruses exhibit limited tropism and replicate poorly in most cell lines. Attachment protein VP4 is a key rotavirus tropism determinant. Previous studies in which human rotaviruses were adapted to cultured cells identified mutations in VP4. However, most such studies were conducted using only a single human rotavirus genotype. In the current study, we serially passaged 50 human rotavirus clinical specimens representing five of the genotypes most frequently associated with severe human disease, each in triplicate, three to five times in primary monkey kidney cells then ten times in the MA104 monkey kidney cell line. From 13 of the 50 specimens, we obtained 25 rotavirus antigen-positive lineages representing all five genotypes, which tended to replicate more efficiently in MA104 cells at late versus early passage. We used Illumina next-generation sequencing and analysis to identify variants that arose during passage. In VP4, variants encoded 28 mutations that were conserved for all P[8] rotaviruses and 12 mutations that were conserved for all five genotypes. These findings suggest there may be a conserved mechanism of human rotavirus adaptation to MA104 cells. In the future, such a conserved adaptation mechanism could be exploited to study human rotavirus biology or efficiently manufacture vaccines. |
