Common vole (Microtus arvalis) and bank vole (Myodes glareolus) derived permanent cell lines differ in their susceptibility and replication kinetics of animal and zoonotic viruses

Binder, Florian; Lenk, Matthias; Weber, Siegfried; Stoek, Franziska; Dill, Veronika; Reiche, Sven; Riebe, Roland; Wernike, Kerstin; Hoffmann, Donata; Ziegler, Ute; Adler, Heiko; Eßbauer, Sandra; Ulrich, Rainer G.

doi:10.1016/j.jviromet.2019.113729

Cited by 17 publications

(17 citation statements)

References 64 publications

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“…Further in vitro studies are needed to determine whether heterologous bank voles evolutionary lineages could support PUUV isolation and/or infection. These future investigations may profit from the recent establishment of permanent bank vole cell lines from different evolutionary lineages of the bank vole (29).…”

Section: Discussionmentioning

confidence: 99%

A novel experimental system reveals immunoregulatory responses as mediators of persistent orthohantavirus infections in a rodent reservoir host

Strandin

Smura

Ahola

et al. 2019

Preprint

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26Orthohantaviruses are globally emerging zoonotic pathogens. Human infections are characterized 27 by an overt immune response that is efficient at counteracting virus replication but can also cause 28 severe tissue damage. In contrast, orthohantavirus infections in rodent reservoir hosts are persistent 29 and asymptomatic. The mechanisms facilitating asymptomatic virus persistence in reservoir hosts 30 are not well understood but could help to guide therapeutic strategies for human infections. Here we 31 report on a study using in vivo and in vitro experiments to investigate immune responses associated 32 with persistent Puumala orthohantavirus (PUUV) infections in the bank vole (Myodes glareolus), its 33 reservoir host. We examined adaptive cellular and humoral responses by quantifying changes in T-34 cell related gene expression in the spleen and immunoglobulin (Ig) responses in blood, respectively. 35 Since existing Vero E6-cell adapted hantavirus isolates have been demonstrated to have lost their 36 wild-type infection characteristics, infections were conducted with a novel PUUV strain isolated on 37 a bank vole cell line. Whole virus genome sequencing revealed that only minor sequence changes 38 occurred during the isolation process, and critically, experimental infections of bank voles with the 39 new isolate resembled natural infections. In vitro infection of bank vole splenocytes with the novel 40 isolate demonstrated that PUUV promotes immunoregulatory responses by inducing interleukin-10, 41 a cytokine strongly associated with chronic viral infections. A delayed virus-specific humoral 42 response occurred in experimentally infected bank voles, which is likely to allow for initial virus 43 replication and the establishment of persistent infections. These results suggest that host 44 immunoregulation facilitates persistent orthohantavirus infections in reservoir hosts. 45 46 Importance 47Orthohantaviruses are a group of global pathogens that regularly spillover from rodent reservoirs 48 into humans and can cause severe disease. Conversely, infections in reservoir hosts do not cause 49 obvious adverse effects. The mechanisms responsible for persistent asymptomatic reservoir 50 3 infections are unknown, and progress has been hindered by the absence of an adequate experimental 51 system. Knowledge on these mechanisms could help provide strategies to treat human infections. 52 We developed and validated an experimental system based on an orthohantavirus isolated in cells of 53 its vole reservoir host. Using animal and cell culture experiments in the reservoir host system, we 54 demonstrated that infection suppresses immunity in the vole reservoir via specific mechanisms, 55 likely allowing the virus to take hold and preventing immune responses that can cause self-damage. 56 57 Introduction 58 Understanding how zoonotic pathogens are maintained and transmitted in nature is critical for 59 efforts to curtail human disease (1). Most investigations have focused on the identification of 60 wild...

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Section: Discussionmentioning

confidence: 99%

A novel experimental system reveals immunoregulatory responses as mediators of persistent orthohantavirus infections in a rodent reservoir host

Strandin

Smura

Ahola

et al. 2019

Preprint

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“…Previous investigations indicated that VeroE6 cell adaptation of PUUV Kazan strain results in the inability of the adapted strain to infect the bank vole reservoir [8]. The recent development of bank vole-derived primary or permanent cell lines may allow the isolation of reservoir-adapted PUUV strains [9][10][11][12].…”

Section: Electronic Supplementary Materialsmentioning

confidence: 99%

“…This endemic region was also again heavily affected by the hantavirus outbreak year 2019 [25]. Here, we aimed to isolate a Central European PUUV strain from bank voles in the district of Osnabrück using standard VeroE6 cells and the recently established Carpathian lineage bank volederived kidney cell line (MGN-2-R [10]). Complete genome determination by shot-gun and hybrid-capture-mediated highthroughput sequencing (HTS) was used to follow the potential adaptation of the PUUV isolates in VeroE6 and reservoir cell lines.…”

Section: Electronic Supplementary Materialsmentioning

confidence: 99%

“…For virus isolation and further infection studies, VeroE6 and bank vole kidney (MGN-2-R; [10]) cells were used in parallel. Virus titration was done on VeroE6 cells only.…”

Section: Cell Linesmentioning

confidence: 99%

“…For immunofluorescence assay (IFA), VeroE6 and MGN-2-R cells were inoculated with 500 µl PUUV Osnabrück/V29 or PUUV Osnabrück/M43 supernatant in DMEM + 5% FCS as described previously [10]. Infected cells were fixed 10 days post infection with a 1:1 mixture of acetone and methanol for 20 min at − 20 °C.…”

Section: Virus Infection and Titrationmentioning

confidence: 99%

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Isolation and characterization of new Puumala orthohantavirus strains from Germany

et al. 2020

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Orthohantaviruses are re-emerging rodent-borne pathogens distributed all over the world. Here, we report the isolation of a Puumala orthohantavirus (PUUV) strain from bank voles caught in a highly endemic region around the city Osnabrück, northwest Germany. Coding and non-coding sequences of all three segments (S, M, and L) were determined from original lung tissue, after isolation and after additional passaging in VeroE6 cells and a bank vole-derived kidney cell line. Different single amino acid substitutions were observed in the RNA-dependent RNA polymerase (RdRP) of the two stable PUUV isolates. The PUUV strain from VeroE6 cells showed a lower titer when propagated on bank vole cells compared to VeroE6 cells. Additionally, glycoprotein precursor (GPC)-derived virus-like particles of a German PUUV sequence allowed the generation of monoclonal antibodies that allowed the reliable detection of the isolated PUUV strain in the immunofluorescence assay. In conclusion, this is the first isolation of a PUUV strain from Central Europe and the generation of glycoprotein-specific monoclonal antibodies for this PUUV isolate. The obtained virus isolate and GPC-specific antibodies are instrumental tools for future reservoir host studies.

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Hantaviruses in a Global Perspective

Krautkrämer

Peintner

Eßbauer

2022

Zoonoses: Infections Affecting Humans and Animals

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Common vole (Microtus arvalis) and bank vole (Myodes glareolus) derived permanent cell lines differ in their susceptibility and replication kinetics of animal and zoonotic viruses

Abstract: Common vole (Microtus arvalis) and bank vole (Myodes glareolus) derived permanent cell lines differ in their susceptibility and replication kinetics of animal and zoonotic viruses

Cited by 17 publications

References 64 publications

A novel experimental system reveals immunoregulatory responses as mediators of persistent orthohantavirus infections in a rodent reservoir host

A novel experimental system reveals immunoregulatory responses as mediators of persistent orthohantavirus infections in a rodent reservoir host

Isolation and characterization of new Puumala orthohantavirus strains from Germany

Hantaviruses in a Global Perspective

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