Recent Advances in Replication and Infection of Human Parvovirus B19

Ganaie, Safder S.; Qiu, Jianming

doi:10.3389/fcimb.2018.00166

Cited by 78 publications

(81 citation statements)

References 133 publications

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“…Gb4 is essential for NS1 expression. In UT7/Epo cells, the expression of NS1 occurs early after nuclear entry (27) and is required for DNA replication and productive infection (28,29). The presence of NS1 mRNA in WT and KO cells was tested at increasing times pi by an NS1-specific quantitative RT-PCR.…”

Section: Generation Of B3galnt1 Ko Ut7/epo Cell Linementioning

confidence: 99%

“…B19V DNA replication and capsid proteins are not detectable in Gb4 KO cells. Following second-strand synthesis, NS1 is required to initiate and maintain virus replication through a rolling hairpin mechanism, which involves the resolution of the inverted terminal repeats (ITRs) (29). Accordingly, the lack of NS1 in KO cells should prohibit virus replication and subsequent steps of the infection, such as capsid protein expression.…”

Section: Generation Of B3galnt1 Ko Ut7/epo Cell Linementioning

confidence: 99%

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Globoside Is Dispensable for Parvovirus B19 Entry but Essential at a Postentry Step for Productive Infection

Bieri

Ros

2019

J Virol

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Globoside (Gb4) is considered the primary receptor of parvovirus B19 (B19V); however, its expression does not correlate well with the attachment and restricted tropism of the virus. The N terminus of VP1 (VP1u) of B19V interacts with an as-yet-unknown receptor required for virus internalization. In contrast to Gb4, the VP1u cognate receptor is expressed exclusively in cells that B19V can internalize. With the aim of clarifying the role of Gb4 as a B19V receptor, we knocked out the gene B3GalNT1 coding for the enzyme globoside synthase in UT7/Epo cells. Consequently, B3GalNT1 transcripts and Gb4 became undetectable in the knockout (KO) cells without affecting cell viability and proliferation. Unexpectedly, virus attachment, internalization, and nuclear targeting were not disturbed in the KO cells. However, NS1 transcription failed, and consequently, genome replication and capsid protein expression were abrogated. The block could be circumvented by transfection with a B19V infectious clone, indicating that Gb4 is not required after the generation of viral double-stranded DNA with resolved inverted terminal repeats. While in wild-type (WT) cells, occupation of the VP1u cognate receptor with recombinant VP1u disturbed virus binding and blocked the infection, antibodies against Gb4 had no significant effect. In a mixed population of WT and KO cells, B19V selectively infected WT cells. This study demonstrates that Gb4 does not have the expected receptor function, as it is dispensable for virus entry; however, it is essential for productive infection, explaining the resistance of the rare individuals lacking Gb4 to B19V infection.IMPORTANCEGloboside has long been considered the primary receptor of B19V. However, its expression does not correlate well with B19V binding and uptake and cannot explain the pathogenesis or the remarkable narrow tissue tropism of the virus. By using a knockout cell line, we demonstrate that globoside does not have the expected function as a cell surface receptor required for B19V entry, but it has an essential role at a postentry step for productive infection. This finding explains the natural resistance to infection associated with individuals lacking globoside, contributes to a better understanding of B19V restricted tropism, and offers novel strategies for the development of antiviral therapies.

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Section: Generation Of B3galnt1 Ko Ut7/epo Cell Linementioning

confidence: 99%

Section: Generation Of B3galnt1 Ko Ut7/epo Cell Linementioning

confidence: 99%

Globoside Is Dispensable for Parvovirus B19 Entry but Essential at a Postentry Step for Productive Infection

Bieri

Ros

2019

J Virol

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“…The clinical use of Parvovirus Assays, p.4 B19V shows a selective tropism for erythroid progenitor cells in the bone marrow, due to the presence of specific receptors, such as the glycolipid globoside and a specific receptor binding the VP1 unique N-terminal domain, and to functional internalization processes [4,5]. In a permissive cellular environment, a coordinated series of macromolecular syntheses occurs [6][7][8]. From the parental single-stranded template, cellular DNA repair synthesis generates a double stranded DNA template, then a first phase transcription mainly produces mRNAs coding for the NS protein, followed by rolling hairpin replication of the genome and extended second phase transcription, mainly producing mRNAs coding for structural VP proteins.…”

Section: The Virus and Its Biologymentioning

confidence: 99%

“…Accumulation of VP proteins eventually leads to the assembly of capsids, encapsidation of progeny single-stranded genomes and release of virions from infected cells. The permissive environment is restricted to cells in the erythroid lineage at differentiation stages ranging from CFU-E to erythroblasts [7], and is critically dependent on Erythropoietin stimulation and hypoxic conditions [6,8], through a signaling cascade eventually leading to formation of a functional replicative complex involving the viral NS and cellular proteins [9]. In productively infected cells, the virus exerts a complex series of effects, including arrest of the cell cycle and induction of apoptosis [10], thus causing a temporary block in erythropoiesis that can manifest as a transient or persistent erythroid aplasia.…”

Section: The Virus and Its Biologymentioning

confidence: 99%

The clinical use of parvovirus B19 assays: recent advances

Gallinella

2018

Expert Review of Molecular Diagnostics

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Parvovirus B19 (B19V), a single-stranded DNA virus in the family Parvoviridae, is a human pathogenic virus, characterized by a selective but not exclusive tropism for erythroid progenitor cells. Widely diffuse, it is responsible for an ample range of clinical manifestations, whose characteristics and outcomes depend on the interplay between the viral properties and the physiological and immune status of the infected individuals. The complexity of virus-host relationship and the diversity of the clinical course of infection pose a diagnostic challenge that may require non-trivial solutions. Areas covered: The review includes an updated description of the course of B19V infection in its complexity and diversity of pathogenetic mechanisms, discusses the consequent requirements for different and appropriated diagnostic approaches, presents the main diagnostic techniques, more recent technical advancements, and their application to the diverse clinical situations. Expert commentary: The complex scenario of the infectious process and the diversity in possible pathogenetic mechanisms make necessary a multi-parametric approach for an accurate and informative laboratory diagnosis of B19V infection, combining as much as possible the molecular detection of viral components, mainly viral DNA, to commonly followed immunological detection of virus-specific antibodies and a critical assessment of laboratory findings.

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“…The minimal replication origin (Ori) of B19V DNA has been identified to be 67 nucleotides (nt), which includes a STAT5-binding element (STAT5BE), a trs or nicking site, two copies of the NS1-binding element (NS1BE), and the putative cellular factorbinding site (CFBE) ( Fig. 2A) (25,27,41,42). Specific cleavage of the ssDNA Ori has been demonstrated in an in vitro nicking assay using the purified B19V NS1 N terminus (aa 1 to 176) and a 5=-end 32 P-labeled oligonucleotide that contain the trs (28).…”

mentioning

confidence: 99%

Endonuclease Activity Inhibition of the NS1 Protein of Parvovirus B19 as a Novel Target for Antiviral Drug Development

Peng

Ganaie

Wang

et al. 2019

Antimicrob Agents Chemother

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Human parvovirus B19 (B19V), a member of the genus Erythroparvovirus of the family Parvoviridae, is a small nonenveloped virus that has a single-stranded DNA (ssDNA) genome of 5.6 kb with two inverted terminal repeats (ITRs). B19V infection often results in severe hematological disorders and fetal death in humans. B19V replication follows a model of rolling hairpin-dependent DNA replication, in which the large nonstructural protein NS1 introduces a site-specific single-strand nick in the viral DNA replication origins, which locate at the ITRs. NS1 executes endonuclease activity through the N-terminal origin-binding domain. Nicking of the viral replication origin is a pivotal step in rolling hairpin-dependent viral DNA replication. Here, we developed a fluorophore-based in vitro nicking assay of the replication origin using the origin-binding domain of NS1 and compared it with the radioactive in vitro nicking assay. We used both assays to screen a set of small-molecule compounds (n ϭ 96) that have potential antinuclease activity. We found that the fluorophore-based in vitro nicking assay demonstrates sensitivity and specificity values as high as those of the radioactive assay. Among the 96 compounds, we identified 8 which have an inhibition of Ͼ80% at 10 M in both the fluorophore-based and radioactive in vitro nicking assays. We further tested 3 compounds that have a flavonoid-like structure and an in vitro 50% inhibitory concentration that fell in the range of 1 to 3 M. Importantly, they also exhibited inhibition of B19V DNA replication in UT7/Epo-S1 cells and ex vivo-expanded human erythroid progenitor cells.

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Recent Advances in Replication and Infection of Human Parvovirus B19

Cited by 78 publications

References 133 publications

Globoside Is Dispensable for Parvovirus B19 Entry but Essential at a Postentry Step for Productive Infection

Globoside Is Dispensable for Parvovirus B19 Entry but Essential at a Postentry Step for Productive Infection

The clinical use of parvovirus B19 assays: recent advances

Endonuclease Activity Inhibition of the NS1 Protein of Parvovirus B19 as a Novel Target for Antiviral Drug Development

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