Molecular characterization of human rotavirus vaccine strain CDC-9 during sequential passages in vero cells

Esona, Mathew D.; Foytich, Kimberly; Wang, Yuhuan; Shin, Gary; Wei, Guiwu; Gentsch, Jon R.; Glass, Roger I.; Jiang, Baoming

doi:10.4161/hv.6.3.10409

Cited by 29 publications

(26 citation statements)

References 52 publications

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“…Substitution at aa 385 was found in two of four studies (27,28). When the VP4 sequence of strain 89-12 (progenitor of Rotarix) was compared to that of the original strain, five substitutions were detected (aa 51, 167, 331, 385, and 695) (27).…”

Section: Discussionmentioning

confidence: 99%

“…When the VP4 sequence of strain 89-12 (progenitor of Rotarix) was compared to that of the original strain, five substitutions were detected (aa 51, 167, 331, 385, and 695) (27). On the other hand, when strain CDC-9 (attenuated live vaccine candidate) was passaged many times, five substitutions (aa 51, 331, 364, 385, and 388) were detected in the VP4 gene (28). In addition to substitution at aa 385, substitution at aa 51 and 331 was confirmed in both studies.…”

Section: Discussionmentioning

confidence: 99%

“…Sequencing analyses between parental (virulent) and serial cell culture-passaged (attenuated) human RV G1P [8] strains were performed for the VP4 gene (26)(27)(28)(29). Substitution at aa 385 was found in two of four studies (27,28).…”

Section: Discussionmentioning

confidence: 99%

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Virulence-Associated Genome Mutations of Murine Rotavirus Identified by Alternating Serial Passages in Mice and Cell Cultures

Tsugawa

Tatsumi

Tsutsumi

2014

J Virol

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Although significant clinical efficacy and safety of rotavirus vaccines were recently revealed in many countries, the mechanism of their attenuation is not well understood. We passaged serially a cell culture-adapted murine rotavirus EB strain in mouse pups or in cell cultures alternately and repeatedly and fully sequenced all 11 genes of 21 virus samples passaged in mice or in cell cultures. Sequence analysis revealed that mouse-passaged viruses that regained virulence almost consistently acquired four kinds of amino acid (aa) substitutions in VP4 and substitution in aa 37 (Val to Ala) in NSP4. In addition, they gained and invariably conserved the 3= consensus sequence in NSP1. The molecular changes occurred along with the acquisition of virulence during passages in mice and then disappeared following passages in cell cultures. Intraperitoneal injection of recombinant NSP4 proteins confirmed the aa 37 site as important for its diarrheagenic activity in mice. These genome changes are likely to be correlated with rotavirus virulence. IMPORTANCESerial passage of a virulent wild-type virus in vitro often results in loss of virulence of the virus in an original animal host, while serial passage of a cell culture-adapted avirulent virus in vivo often gains virulence in an animal host. Actually, live attenuated virus vaccines were originally produced by serial passage in cell cultures. Although clinical efficacy and safety of rotavirus vaccines were recently revealed, the mechanism of their attenuation is not well understood. We passaged serially a murine rotavirus by alternating switch of host (mice or cell cultures) repeatedly and sequenced the eleven genes of the passaged viruses to identify mutations associated with the emergence or disappearance of virulence. Sequence analysis revealed that changes in three genes (VP4, NSP1, and NSP4) were associated with virulence in mice. Intraperitoneal injection of recombinant NSP4 proteins confirmed its diarrheagenic activity in mice. These genome changes are likely to be correlated with rotavirus virulence. R otaviruses, which form one genus within the family Reoviridae, are divided into at least seven species/groups (A to G/H) (1, 2). Although group A to C rotaviruses have been detected in humans with diarrhea, group A rotavirus is the single most important etiologic agent causing severe diarrhea in infants and young children worldwide, resulting in approximately 453,00 deaths (37% of deaths attributable to diarrhea and 5% of all deaths) among children Ͻ5 years of age in 2008 (3). In the United States alone, rotavirus (RV) infections are estimated to cause approximately 20 to 30 deaths, 50,000 to 67,000 hospitalizations, 390,000 to 410,000 physician visits, and a more than $890 million to $1 billion societal cost annually (4, 5). Thus, the introduction of a RV vaccine capable of alleviating this enormous health burden has been an important global public health goal.The RV genome consisting of 11 segments of double-stranded RNA (dsRNA) encodes six structural protein...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Virulence-Associated Genome Mutations of Murine Rotavirus Identified by Alternating Serial Passages in Mice and Cell Cultures

Tsugawa

Tatsumi

Tsutsumi

2014

J Virol

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“…The strain is a single gene reassortant with the VP3 gene derived naturally from a G2P4 virus and the other 10 genes from a G1P8 virus, the most common genotype throughout the world. 12 The strain was selected by serial passages and plaque commentAry commentAry or very low levels of IgG and neutralizing activity in pre-bled and post-dose 1 sera. Animals that received two doses of IRV developed significantly higher titers of IgG (mean = 3,040) and neutralizing activity against the two human strainshomotypic Wa (mean = 320) and heterotypic MW333 (mean = 176).…”

Section: Does a Monovalent Inactivated Human Rotavirus Vaccine Inducementioning

confidence: 99%

Does a monovalent inactivated human rotavirus vaccine induce heterotypic immunity?

Jiang

Wang

Glass

2013

Human Vaccines & Immunotherapeutics

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“…1,2 This strain possesses desired biophysical, biochemical and virological characteristics for an IRV, as it grows to high titer (up to 10 8 ffu/ml) in Vero cells and maintains predominant triple-layered particles (>95%) during upstream manufacture and downstream purification processes. Inactivated CDC-9 strain alone when administered to skin using a microneedle patch was highly immunogenic even at a fractional dose of an intramuscular dose in mice.…”

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confidence: 99%

Inactivated human rotavirus vaccine induces heterotypic antibody response: Correction and development of IgG avidity assay

Velásquez

Wang²,

Jiang³

2015

Human Vaccines & Immunotherapeutics

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Molecular characterization of human rotavirus vaccine strain CDC-9 during sequential passages in vero cells

Cited by 29 publications

References 52 publications

Virulence-Associated Genome Mutations of Murine Rotavirus Identified by Alternating Serial Passages in Mice and Cell Cultures

Virulence-Associated Genome Mutations of Murine Rotavirus Identified by Alternating Serial Passages in Mice and Cell Cultures

Does a monovalent inactivated human rotavirus vaccine induce heterotypic immunity?

Inactivated human rotavirus vaccine induces heterotypic antibody response: Correction and development of IgG avidity assay

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