Persistent Reptarenavirus and Hartmanivirus Infection in Cultured Boid Cells

Lintala, Annika; Szirovicza, Leonóra; Kipar, Anja; Hetzel, Udo; Hepojoki, Jussi

doi:10.1128/spectrum.01585-22

Cited by 5 publications

(23 citation statements)

References 44 publications

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“…The frequent reptarenavirus coinfections in boa constrictors with BIBD (26,27) suggest that reptarenavirus S and L segments might re-assort rather freely (27). As studied above, the S to L segment ratio in the supernatant appeared to differ from the ratio observed in the infected cells in our earlier study (18). We thus studied whether coinfection would alter the ratio of S and L segments released.…”

Section: Coinfection Does Not Affect the Ratio Of L And S Segments Re...mentioning

confidence: 77%

“…100,000,000 to 100,000 copies/reaction) of the respective control RNAs in duplicate. The primers and probes as well as the qRT-PCR protocol for quantifying the S and L segments and housekeeping gene were as described (18). In brief, the reactions with Taqman Fast Virus 1-step master mix (ThermoFisher scientific) were half of the recommended 20 μl volume, and the reaction components were: 2.5 μl of the master mix, 2.5 μl of template, 0.25 μl of each primer (20 μM stock), 0.125 μl of the probe (20 μM stock), and 4.375 μl of water.…”

Section: Rna Extraction and Quantitative Reverse Transcription-polyme...mentioning

confidence: 99%

“…When transmitted to humans, mammarenaviruses can cause severe, even fatal disease (11) whereas the members of other arenavirus genera have not been associated to human disease. Reptarenaviruses cause boid inclusion body disease (BIBD) (12)(13)(14)(15)(16), a disease described in private and zoological collections of captive boid snakes since the 1970s (17) and recently also in free-ranging Costa Rican boa constrictors (18). The formation of eosinophilic intracytoplasmic inclusion bodies (IBs) in different cell types of the affected snakes is characteristic to BIBD (17).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, experimental infection (12,15), vertical transmission (19) and screening studies performed on snake collections (20)(21)(22)(23)(24)(25) suggest that reptarenaviruses establish a persistent infection at least in boa constrictors. We recently demonstrated that both reptarena-and hartmaniviruses can establish a persistent infection in cultured boid kidney cells (18).…”

Section: Introductionmentioning

confidence: 99%

“…We hypothesize that the frequently occurring reptarenavirus coinfections in captive snakes with BIBD could be a result of superinfection of persistently reptarenavirus-infected snakes with novel S and L segments upon introduction of new snakes into a collection. To mimic such a scenario in cell culture, we tested if the persistently reptarenavirus infected cell cultures described in our previous study (18) could be superinfected with closely or distantly related reptarenaviruses, or with a hartmanivirus. For the study, we employed five reptarenavirus and one hartmanivirus isolates, and the three persistently infected boa constrictor kidney cell lines of the earlier study.…”

Section: Introductionmentioning

confidence: 99%

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Reptarenavirus Co- and Superinfection in Cell Culture Sheds Light on the S and L Segment Accumulation in Captive Snakes

Lintala

Szirovicza

Kipar

et al. 2022

Preprint

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Boid inclusion body disease (BIBD) caused by reptarenaviruses affects collections of captive constrictor snakes worldwide. The disease manifests by formation of cytoplasmic inclusion bodies (IBs) in various tissues. Curiously, a snake with BIBD most often carries more than a single pair of genetically distinct reptarenavirus S and L segments, and the tissues of an infected individual can show variation in the variety of different S and L segment species. The role of reptarenavirus coinfection in development of BIBD remains unknown, and it is unclear how the infection affects the susceptibility to reptarenavirus superinfection or to secondary infection by other agents. Because cell culture studies with mammarenaviruses have demonstrated persistently infected cultures to resist superinfection by genetically similar viruses, we hypothesized that coinfection would only occur if the infecting viruses were of two different species. To study the hypothesis, we employed boa constrictor kidney- and brain-derived cell cultures to perform a set of co- and superinfection experiments with five reptarenavirus and one hartmanivirus isolates. Analysis of viral RNA released from coinfected cells using qRT-PCR did not demonstrate evident competition between reptarenaviruses of the same or different species in the boa constrictor kidney-derived cell line. The experiments on the brain-derived cell line revealed considerable differences in the replication ability of the reptarenaviruses tested, suggesting varying tissue specificity or target cell spectra for reptarenavirus species. Finally, experiments on persistently reptarenavirus infected cell lines showed reduced replication of closely related reptarenaviruses while the replication of reptarenaviruses from different species appeared unaffected.

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Section: Coinfection Does Not Affect the Ratio Of L And S Segments Re...mentioning

confidence: 77%

Section: Rna Extraction and Quantitative Reverse Transcription-polyme...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Reptarenavirus Co- and Superinfection in Cell Culture Sheds Light on the S and L Segment Accumulation in Captive Snakes

Lintala

Szirovicza

Kipar

et al. 2022

Preprint

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Cell culture co- and superinfection experiments suggest that transmission during captivity contributes to the presence of reptarenavirus S and L segment swarms in boid inclusion body disease-positive snakes

Lintala,

Szirovicza,

Sander

et al. 2024

Journal of General Virology

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Boid inclusion body disease (BIBD) caused by reptarenaviruses affects captive constrictor snake collections worldwide. The disease manifests by the formation of cytoplasmic inclusion bodies in various tissues. Curiously, a snake with BIBD nearly always carries a swarm of reptarenavirus small and large segments rather than a single pair, and the composition of the swarm can vary between tissues. The role of reptarenavirus coinfections in BIBD pathogenesis remains unknown, and it is unclear whether reptarenavirus infection affects the susceptibility to superinfection or to secondary infections. For mammarenaviruses, co- and/or superinfection can occur if the infecting viruses are genetically divergent enough, and we hypothesized reptarenaviruses to behave similarly. To study this hypothesis, we employed boa constrictor kidney- and brain-derived cell cultures to perform a set of co- and superinfection experiments with one hartmanivirus and five reptarenavirus isolates. While all tested viruses replicated well in the boid kidney cells, experiments on the brain-derived cells showed differences in the replication efficacy between the viruses, suggesting that reptarenaviruses could differ in their target cell spectra. The quantification of viral RNA released from infected cells as a proxy for virus replication did not reveal overt differences between mono- and coinfections. Passaging of coinfected cell cultures revealed that one of the reptarenavirus isolates requires a coinfecting reptarena- or hartmanivirus to establish a persistent infection. Superinfection experiments on persistently reptarenavirus-infected cell lines suggested some interference between genetically similar viruses. We hypothesized that such interference would be mediated by the viral Z protein (ZP) specifically locking the genetically similar viral polymerase in a catalytically inactive state. Curiously, experiments on ZP-expressing cell lines indicated ZP overexpression not to significantly affect the amount of released viral RNA. Our experiments showed very little co- or superinfection interference between genetically dissimilar reptarenaviruses, reflecting the naturally occurring reptarenavirus coinfections in snakes with BIBD.

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Reptarenavirus S Segment RNA Levels Correlate with the Presence of Inclusion Bodies and the Number of L Segments in Snakes with Reptarenavirus Infection—Lessons Learned from a Large Breeding Colony

et al. 2023

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Boid inclusion body disease (BIBD) is caused by reptarenavirus infection, and while reptarenavirus nucleoprotein is the main component of the inclusion bodies (IBs) characteristic of BIBD, not all reptarenavirus-infected snakes demonstrate IBs in their cells. Identification of infected individuals is critical for controlling the spread of the disease; however, the genetic divergence of reptarenaviruses complicates reverse transcription-PCR (RT-PCR)-based diagnostics.

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Persistent Reptarenavirus and Hartmanivirus Infection in Cultured Boid Cells

Abstract: Mammarenaviruses cause a persistent infection in their natural rodent and bat hosts. Reptarenaviruses cause boid inclusion body disease (BIBD) in constrictor snakes, but it is unclear whether snakes are the natural host of these viruses.

Cited by 5 publications

References 44 publications

Reptarenavirus Co- and Superinfection in Cell Culture Sheds Light on the S and L Segment Accumulation in Captive Snakes

Reptarenavirus Co- and Superinfection in Cell Culture Sheds Light on the S and L Segment Accumulation in Captive Snakes

Cell culture co- and superinfection experiments suggest that transmission during captivity contributes to the presence of reptarenavirus S and L segment swarms in boid inclusion body disease-positive snakes

Reptarenavirus S Segment RNA Levels Correlate with the Presence of Inclusion Bodies and the Number of L Segments in Snakes with Reptarenavirus Infection—Lessons Learned from a Large Breeding Colony

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