Homologous interference mediated by defective interfering influenza virus derived from a temperature-sensitive mutant of influenza virus

Nayak, Debi P.; Tobita, Kiyotake; Janda, Jozef; Davis, Alan R.; De, Barun K.

doi:10.1128/jvi.28.1.375-386.1978

Cited by 93 publications

(75 citation statements)

References 31 publications

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“…The presence of defective influenza genomes has been well characterized in cell cultures (2–7) and animal models (25). Influenza DVGs have also been observed in clinical human H1N1 samples (8).…”

Section: Discussionmentioning

confidence: 99%

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The Dynamics of Influenza A H3N2 Defective Viral Genomes from a Human Challenge Study

Martin

Kaul

et al. 2019

Preprint

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248 words 17 Importance: 135 words 18 Main text: 4,747 words 19 Importance 39The evolution of influenza virus, in terms of single nucleotide variants and the reassortment of 40 gene segments, has been studied in detail. However, influenza is known to generate defective 41 viral genomes (DVGs) during replication, and little is known about how these genomes evolve 42 both within hosts and at the population level. Studies in animal models have indicated that 43 prophylactically or therapeutically administered DVGs can impact patterns of disease 44 progression. However, the formation of naturally-occurring DVGs, their evolutionary dynamics, 45and their contribution to disease severity in human hosts is not well understood. Here, we 46 identify the formation of de novo DVGs in samples from human challenge studies throughout the 47 course of infection. We analyze their evolutionary trajectories, revealing the important role of 48 genetic drift in shaping DVG populations during acute infections with well-adapted viral strains. 49 of the samples analyzed and were most common in the PB2, PB1, and PA gene segments. A 73 limitation of this study, however, is that it offered only a cross-sectional view of IAV DVG 74 populations. While it has been shown that Sendai virus DVG populations expand during the first 75 12 hours of infection in a mouse model (25), the evolution of IAV DVG populations within a 76 human host over the course of an infection has not been well characterized. 77Here, we report an analysis of IAV DVG populations identified from deep sequencing 78 data taken over the course of infection during two longitudinal human challenge studies with 79 different treatment cohorts. We observe the generation of de novo DVGs in nearly all subjects, 80 primarily in the polymerase gene segments (PB2, PB1, and PA). DVG populations were highly 81 variable over time in DVG species composition as well as in DVG species relative abundance. 82Over the course of infection, individual DVG species were observed to arise, fluctuate in 83 abundance, as well as disappear from the DVG population. Overall, we found no trend towards 84 decreasing diversity of DVG populations or towards shorter DVG species during the five days 85 post challenge, likely due to the dominance of stochastic effects. Furthermore, we were unable to 86 detect an association between DVG levels and peak viral titers, potentially due to the negative 87 feedback between DVG and wild-type virus. Similarly, higher DVG levels were not associated 88 with more severe symptoms. This study helps to illustrate the stochastic dynamics of DVG 89 populations within a host during acute infection with a well-adapted viral strain, a scenario under 90 which fitness variation in the wild-type virus population is expected to be relatively small. 91 Materials and Methods 92Ethics statement. The procedures followed in the human challenge studies were in accordance 93 with the Declaration of Helsinki. The studies were approved by the institutional review boards 94 (IRBs) of Duke Univer...

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Section: Discussionmentioning

confidence: 99%

“…Influenza defective viral genomes (DVGs) were first reported by von Magnus (1) and have since been characterized in vivo during high multiplicity of infection (MOI) passage studies (2–7) as well as from clinical human samples (8, 9). DVGs are classified as viral genomes harboring mutations which render them incapable of self-replication.…”

Section: Introductionmentioning

confidence: 99%

The Dynamics of Influenza A H3N2 Defective Viral Genomes from a Human Challenge Study

Martin

Kaul

et al. 2019

Preprint

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“…With influenza, von Magnus (26) originally described the formation of noninfectious virus by successive high-multiplicity undiluted passages of virus in embryonated eggs. Recently we and others have reported the presence of small deleted RNA segments (DI RNA) in DI influenza viral preparations (5,17,19,20,22). Furthermore, based on several lines of evidence (albeit indirect), we have postulated that, as in the case of other viruses, these DI RNA molecules are responsible for DI virus-mediated interference.…”

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confidence: 94%

Defective Influenza Viral Ribonucleoproteins Cause Interference

Janda

Nayak

1979

J Virol

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Ribonucleoproteins (RNPs) isolated from infectious and defective interfering (DI) influenza virus (WSN) contained three major RNP peaks when analyzed in a glycerol gradient. Peak I RNP was predominant in infectious virus but was greatly reduced in DI virus preparations. Conversely, peak III RNP was elevated in DI virus, suggesting a large increase in DI RNA in this fraction. Labeled [32P]RNA was isolated from each RNP region and analyzed by electrophoresis on polyacrylamide gels. Peak I RNP contained primarily the polymerase and some HA genes, peak II contained some HA gene but mostly the NP and NA genes, and peak III contained the M and NS genes. In addition, peak III RNP from DI virus also contained the characteristic DI RNA segments. Interference activity of RNP fractions isolated from infectious and DI virus was tested using infectious center reduction assay. RNP peaks (I, II, and III) from infectious virus did not show any interference activity, whereas the peak III DI RNP caused a reduction in the number of infectious centers as compared to controls. Similar interference was not demonstrable with peak I RNP of DI virus nor with any RNP fractions from infectious virus alone. The interference activity of RNP fractions was RNase sensitive, suggesting that the DI RNA contained in DI RNPs was the interfering agent, and dilution experiments supported the conclusion that a single DI RNP could cause interference. The interfering RNPs were heterogeneous, and the majority migrated slower than viral RNPs containing M and NS genes. These results suggest that DI RNP (or DI RNA) is also responsible for interference in segmented, negative-stranded viruses.

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“…They interfere with the replication of homologous infectious viruses at the level of synthesis of virus-specified RNA (7). Nayak et al (1978) reported that DI particles of a temperature-sensitive mutant of WSN influenza virus, produced by successive undiluted passage in MDBK cells, did not inhibit the synthesis of cRNA of standard virus but inhibited specifically vRNA synthesis. In the present study, it seems most likely that noninfectious virus particles contained in the von Magnus virus preparation inhibited the synthesis of vRNA by coexisting infectious virus.…”

Section: Discussionmentioning

confidence: 99%

“…The other is a slowly sedimenting peak (fractions 11-15) whose radioactivity was as much as that in fractions 11-15 of standard virus-infected cells. The von Magnus incomplete virus is known to have small RNA molecules that are not present in the standard influenza virus (7,13,14,16). Small-sized RNPs containing this kind of RNA molecules may exist in these fractions although further experiments are needed to confirm this possiblity.…”

Section: Intracellular Distribution If Viral Rnp and Soluble Npmentioning

confidence: 99%

Analysis of Nuclear Accumulation of Influenza NP Antigen in von Magnus Virus‐Infected Cells

Maeno

Aoki

Hamaguchi

et al. 1981

Microbiology and Immunology

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When 1-50-4 cells were infected with von Magnus virus derived from influenza AJRIJ5+ virus by four successive undiluted passages in chick embryos, virus-specific proteins were synthesized but production of infectious virus was inhibited. In these cells the synthesis of viral RNA was suppressed and the nucleoprotein (NP) antigen was found predominantly in the nucleus in contrast to standard virus-infected cells in which the antigen was distributed throughout the whole cell. The intracellular location and migration of NP were determined by isotope labeling and sucrose gradient centrifugation of subcellular fractions. In standard virus-infected cells NP polypeptide was present predominantly in the cytoplasm in the form of viral ribonucleoprotein (RNP) and intranuclear RNP was detected in reduced amounts. In contrast, in von Magnus virus-infected cells NP polypeptide was present predominantly in the nucleus in a non assembled, soluble form and the amount of cytoplasmic RNP was considerably reduced. After short-pulse labeling NP was detected exclusively in the cytoplasm in a soluble form and after a chase a large proportion of such soluble NP was seen in the nucleus. It is suggested that a large proportion of the NP synthesized in von Magnus virus-infected cells is not assembled into cytoplasmic RNP because of the lack of available RNA and the NP migrated into the nucleus and remained there.Canavanine, an arginine analog, is a potent inhibitor of the growth of influenza virus (11). It inhibits the synthesis of viral RNA and the formation of viral RNP.Under these conditions, most of the NP is found in the nucleus in a nonassembled, soluble form, in contrast to untreated cells in which a large amount of NP is distributed in the cytoplasm in the form of RNP. Influenza RNP consists of multiple molecules ofNP and one segment of virion RNA (vRNA) or cRNA complementary to vRNA (2, 17). We have suggested the possibility that the NP synthesized in the presence of canavanine is not assembled into cytoplasmic RNP because of the lack of available RNA and that the NP readily migrates into the nucleus and ac-1 Present address: Gamagori-Fukashi Hospital, Gamagori, Aichi 433. 283

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Homologous interference mediated by defective interfering influenza virus derived from a temperature-sensitive mutant of influenza virus

Cited by 93 publications

References 31 publications

The Dynamics of Influenza A H3N2 Defective Viral Genomes from a Human Challenge Study

The Dynamics of Influenza A H3N2 Defective Viral Genomes from a Human Challenge Study

Defective Influenza Viral Ribonucleoproteins Cause Interference

Analysis of Nuclear Accumulation of Influenza NP Antigen in von Magnus Virus‐Infected Cells

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