Whole-Genome Sequences of Zika Virus FLR Strains after Passage in Vero or C6/36 Cells

Moser, Lindsey A.; Oldfield, Lauren M.; Fedorova, Nadia; Puri, Vinita; Shrivastava, Susmita; Amedeo, Paolo; Isom, Richard; Hu, Lihui; Durbin, Alan; Rocchi, Iara; Williams, Torrey; Tan, Gene S.; Shabman, Reed S.; Bernard, Kristen A.; Pickett, Brett E.

doi:10.1128/genomea.01528-17

Cited by 3 publications

(5 citation statements)

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“…We examined the genomes of all lineages of the plaque purified pass 3 biological clones by next-generation sequencing as previously described [ 38 ]. The results of this analysis are shown in Table 3 .…”

Section: Resultsmentioning

confidence: 99%

“…RNA samples were sequenced as described in our previous report [ 38 ] using the RNA sequence-independent single-primer amplification (SISPA) method [ 39 , 40 ]. All samples were sequenced using 300 bp paired-end reads on an Illumina MiSeq instrument with a subset of samples sequenced on an Ion Torrent instrument.…”

Section: Methodsmentioning

confidence: 99%

“…All samples were sequenced using 300 bp paired-end reads on an Illumina MiSeq instrument with a subset of samples sequenced on an Ion Torrent instrument. Read assembly was performed as previously described [ 38 ]. Briefly, after reads were deconvoluted and trimmed, the contigs were mapped to the most appropriate ZIKV genome.…”

Section: Methodsmentioning

confidence: 99%

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Growth and adaptation of Zika virus in mammalian and mosquito cells

et al. 2018

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The recent emergence of Zika virus (ZIKV) in the Americas coincident with increased caseloads of microcephalic infants and Guillain-Barre syndrome has prompted a flurry of research on ZIKV. Much of the research is difficult to compare or repeat because individual laboratories use different virus isolates, growth conditions, and quantitative assays. Here we obtained three readily available contemporary ZIKV isolates and the prototype Ugandan isolate. We generated stocks of each on Vero mammalian cells (ZIKVmam) and C6/36 mosquito cells (ZIKVmos), determined titers by different assays side-by-side, compared growth characteristics using one-step and multi-step growth curves on Vero and C6/36 cells, and examined plaque phenotype. ZIKV titers consistently peaked earlier on Vero cells than on C6/36 cells. Contemporary ZIKV isolates reached peak titer most quickly in a multi-step growth curve when the amplifying cell line was the same as the titering cell line (e.g., ZIKVmam titered on Vero cells). Growth of ZIKVmam on mosquito cells was particularly delayed. These data suggest that the ability to infect and/or replicate in insect cells is limited after growth in mammalian cells. In addition, ZIKVmos typically had smaller, more homogenous plaques than ZIKVmam in a standard plaque assay. We hypothesized that the plaque size difference represented early adaptation to growth in mammalian cells. We plaque purified representative-sized plaques from ZIKVmos and ZIKVmam. ZIKVmos isolates maintained the initial phenotype while plaques from ZIKVmam isolates became larger with passaging. Our results underscore the importance of the cells used to produce viral stocks and the potential for adaptation with minimal cell passages. In addition, these studies provide a foundation to compare current and emerging ZIKV isolates in vitro and in vivo.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Growth and adaptation of Zika virus in mammalian and mosquito cells

et al. 2018

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“…All passage two ZIKV isolates were also deep sequenced, and resulted in no amino acid differences from passage one isolates. In studies done by others, ZIKV-FLR was passaged up to eight times in Vero or C6/36 cells, and resulted in 99.999% nucleotide sequence identity across 26 different FLR-derived isolates [ 38 ]. Furthermore, all three of the ZIKV isolates used in this study replicate similarly in C6/36 mosquito cells, commonly used for flavivirus propagation ( S1 Fig ).…”

Section: Methodsmentioning

confidence: 99%

IP-10 and CXCR3 signaling inhibit Zika virus replication in human prostate cells

et al. 2020

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Our previous studies have shown that Zika virus (ZIKV) replicates in human prostate cells, suggesting that the prostate may serve as a long-term reservoir for virus transmission. Here, we demonstrated that the innate immune responses generated to three distinct ZIKV strains (all isolated from human serum) were significantly different and dependent on their passage history (in mosquito, monkey, or human cells). In addition, some of these phenotypic differences were reduced by a single additional cell culture passage, suggesting that viruses that have been passaged more than 3 times from the patient sample will no longer reflect natural phenotypes. Two of the ZIKV strains analyzed induced high levels of the IP-10 chemokine and IFNγ in human prostate epithelial and stromal mesenchymal stem cells. To further understand the importance of these innate responses on ZIKV replication, we measured the effects of IP-10 and its downstream receptor, CXCR3, on RNA and virus production in prostate cells. Treatment with IP-10, CXCR3 agonist, or CXCR3 antagonist significantly altered ZIKV viral gene expression, depending on their passage in cells of relevant hosts (mosquito or human). We detected differences in gene expression of two primary CXCR3 isoforms (CXCR3-A and CXCR3-B) on the two cell types, possibly explaining differences in viral output. Lastly, we examined the effects of IP-10, agonist, or antagonist on cell death and proliferation under physiologically relevant infection rates, and detected no significant differences. Although we did not measure protein expression directly, our results indicate that CXCR3 signaling may be a target for therapeutics, to ultimately stop sexual transmission of this virus.

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“…O vírus Zika (VZIK) é um arbovirus, sendo este grupo de vírus transmitido por artrópodes (Casseb et al,2013). Este vírus vem tornando-se um dos arbovírus de maior atenção da comunidade cientifica devido as manifestações causadas por sua infecção, principalmente envolvendo neonatos, onde foram descritas crianças com microcefalia, proveniente de uma afinidade do vírus a infectar células neurais progenitoras prejudicando o crescimento do cérebro, além de poder causar a síndrome Diante disso, este trabalho busca demonstrar através da técnica de transcriptoma o perfil de expressão dos mRNAs dos testículos durante a infecção pelo Vírus Zika, na tentativa de elucidar a influencia da infecção sobre os processos gênicos do órgão, visto que foram descritos caso de lesões, atrofia e até infertilidade (MOSER, et al 2018).…”

Section: Introductionunclassified