Isolation of Hokkaido virus, genus Hantavirus, using a newly established cell line derived from the kidney of the grey red-backed vole (Myodes rufocanus bedfordiae)

Hantaviruses belong to the family Bunyaviridae and are maintained in wild rodents. Although Vero E6 cells, which originate from African green monkey kidney, are used widely in hantavirus research, isolation of hantaviruses from this cell line is difficult. To develop an efficient method of propagation and isolation of hantaviruses we established a novel cell line, MRK101, derived from the kidney of the grey red-backed vole (Myodes rufocanus bedfordiae), the natural host of Hokkaido virus (HOKV). The MRK101 cells showed a significantly higher susceptibility to Puumala virus (PUUV) hosted by Myodes glareolus than Vero E6 cells. Viral nucleocapsid protein in PUUV-infected MRK101 cells was detected earlier than in Vero E6 cells, and the viral titre in the culture fluid of MRK101 cells was higher than that of Vero E6 cells during the early phase of infection. In contrast, MRK101 cells showed no susceptibility to Hantaan virus. HOKV, which has not been isolated to date, was isolated successfully using MRK101 cells. Moreover, the newly isolated HOKV was successfully propagated in MRK101, but not Vero E6, cells. Phylogenic analyses of the S (small), M (medium) and L (large) segment sequences revealed that HOKV is related most closely to PUUV, but is distinct from other hantaviruses. These data suggest that the MRK101 cell line is a useful tool for the isolation and propagation of hantaviruses. Moreover, this is (to our knowledge) the first report of hantavirus isolation in a cell line that originated from the natural host.

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First isolation and genetic characterization of Puumala orthohantavirus strains from France

10.1101/2020.08.27.270181 ◽

2020 ◽

Cited By ~ 2

Author(s):

Johann Vulin ◽

Séverine Murri ◽

Sarah Madrières ◽

Maxime Galan ◽

Caroline Tatard ◽

...

Keyword(s):

Bank Vole ◽

Endemic Area ◽

High Throughput Sequencing ◽

Hemorrhagic Fever ◽

Amino Acid Sequences ◽

Myodes Glareolus ◽

Wild Rodents ◽

Bank Voles ◽

A Cell ◽

The Impact

AbstractPuumala orthohantavirus (PUUV) causes a mild form of hemorrhagic fever with renal syndrome (HFRS) named nephropathia epidemica (NE), regularly diagnosed in Europe. France represents the Western frontier of NE expansion in Europe with two distinct areas: the endemic area (Northeast) where PUUV circulates in rodent populations and where many cases of NE are detected in humans and non-endemic area (Southwest) where the virus is not detected and only a few human cases have been reported. The country is a pertinent target to study factors that influence the evolution of PUUV distribution. In this study, we describe for the first time the isolation of two PUUV strains from two distinct French geographical areas: Ardennes (endemic area) and Loiret (non-endemic area). To isolate PUUV efficiently, we selected wild rodents (Myodes glareolus, the specific reservoir of PUUV) from these areas that were seronegative for anti-PUUV IgG (ELISA) but associated with viral RNA load in lung (qRT-PCR). With this design, we are able to cultivate and maintain these two strains in VeroE6 cells but also to propagate efficiently and rapidly both strains in a bank vole colony. Complete coding sequences of S and M segments were determined by Sanger sequencing of RNA extracted from positive bank voles (naturally and experimentally infected) and from supernatant of Vero E6. For the M segment, nucleotidic sequences were 100% identical for both strains. For the S segment, the amino acid sequences from each strain revealed one mismatch between sequences obtained from tissue and from supernatant, revealing a “bank vole” and a “cell” signature. High throughput sequencing confirmed Sanger results, and provided a better assessment of the impact of isolation methods on intra-host viral diversity.

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THE CELL CYCLE OF AN ESTABLISHED CELL LINE OF THE MOSQUITO AEDES AEGYPTI

Canadian Journal of Genetics and Cytology ◽

10.1139/g78-042 ◽

1978 ◽

Vol 20 (3) ◽

pp. 373-376 ◽

Cited By ~ 1

Author(s):

Amy S. Baim ◽

Asit B. Mukherjee

Keyword(s):

Cell Cycle ◽

Aedes Aegypti ◽

Cell Line ◽

Mosquito Species ◽

Total Cell ◽

Established Cell Line ◽

Cell Cycle Time ◽

A Cell ◽

High Resolution Autoradiography ◽

Established Cell

The duration of the cell cycle and its four phases was determined for a cell line of the mosquito, Aedes aegypti (L.), using high-resolution autoradiography. The total cell cycle time is 12.5 h, with G1 comprising 1.66 h, S – 4.5 h, M – 3 h, and G2 3.33 h. These results are compared with those of other mosquito species.

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Molecular cloning of the breakpoint for 3q27 translocation in B-cell lymphomas and leukemias

Blood ◽

10.1182/blood.v83.1.217.bloodjournal831217 ◽

1994 ◽

Vol 83 (1) ◽

pp. 217-222 ◽

Cited By ~ 3

Author(s):

T Miki ◽

N Kawamata ◽

A Arai ◽

K Ohashi ◽

Y Nakamura ◽

...

Keyword(s):

B Cell ◽

Cell Line ◽

Established Cell Line ◽

Chain Gene ◽

Light Chain Gene ◽

Normal Tissues ◽

A Cell ◽

Established Cell ◽

Hematopoietic Cell Lines

Reciprocal exchanges between chromosomal region 3q27 and three loci of the Ig genes have been reported in cases of B-cell type non-Hodgkin's lymphoma. We have cloned a region containing a breakpoint junction of 3q27 from a cell line established from a patient with Burkitt's lymphoma carrying t(3;22)(q27;q11). The region cloned was shown to contain an Ig lambda light chain gene fused to a gene on chromosome 3q27. This finding was subsequently confirmed by fluorescence in situ hybridization. Extra nucleotides were present at the joining site. The heptamer-like and nonamer-like sequences separated by an intervening 24 bp were present in the region corresponding to the breakpoint of 3q27, suggesting that a misrecombination in Ig gene rearrangement may be involved in the translocation. Southern blot analysis with a 3q27- specific probe showed rearrangements in three additional patients with B-cell malignancies with the t(3;14)(q27;q32). The breakpoints of all four cases clustered within a limited 3-kb region on chromosome 3q27. The region of 3q27 involved in the translocation was designated as the BCL5 locus. The transcripts from the BCL5 locus were detected in normal tissues and hematopoietic cell lines, and the increased expression of transcript of aberrant size was detected in the established cell line carrying t(3;22). These observations suggest that a gene located at 3q27 is involved in the translocation and that its deregulation plays a role in the malignant transformation of B cells.

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Alpha/Beta Interferon (IFN-α/β)-Independent Induction of IFN-λ1 (Interleukin-29) in Response to Hantaan Virus Infection

Journal of Virology ◽

10.1128/jvi.00717-10 ◽

2010 ◽

Vol 84 (18) ◽

pp. 9140-9148 ◽

Cited By ~ 41

Author(s):

Malin Stoltz ◽

Jonas Klingström

Keyword(s):

Virus Infection ◽

Signaling Pathways ◽

Cell Line ◽

A549 Cells ◽

Hantaan Virus ◽

Poly I:C ◽

Type I ◽

Type I Ifn ◽

Type I Ifns ◽

A Cell

ABSTRACT Type III interferons ([IFNs]IFN-λ and interleukin-28 and -29 [IL-28/29]) are recently recognized cytokines with innate antiviral effects similar to those of type I IFNs (IFN-α/β). Like IFN-α/β, IFN-λ-expression can be induced by viruses, and it is believed that type I and III IFNs are regulated in the same manner. Hantaviruses are weak IFN-α/β inducers and have surprisingly been shown to activate IFN-α/β-independent IFN-stimulated gene (ISG) expression. Here, we show that in Hantaan virus (HTNV)-infected human epithelial A549 cells, induction of IFN-λ1 preceded induction of MxA and IFN-β by 12 and 24 h, respectively, and IFN-α was not induced at all. Furthermore, induction of IFN-λ1 and MxA was observed in HTNV-infected African green monkey epithelial Vero E6 cells, a cell line that cannot produce type I IFNs, clearly showing that HTNV can induce IFN-λ1 and ISGs in the complete absence of IFN-α/β. In HTNV-infected human fibroblast MRC-5 cells, which lack the IFN-λ receptor, induction of MxA coincided in time with IFN-β-induction. UV-inactivated HTNV did not induce any IFNs or MxA in any cell line, showing that activation of IFN-λ1 is dependent on replicating virus. Induction of both IFN-β and IFN-λ1 in A549 cells after poly(I:C)-stimulation was strongly inhibited in HTNV-infected cells, suggesting that HTNV can inhibit signaling pathways used to simultaneously activate types I and III IFNs. In conclusion, we show that HTNV can cause type I IFN-independent IFN-λ1 induction and IFN-λ1-specific ISG induction. Importantly, the results suggest the existence of specific signaling pathways that induce IFN-λ1 without simultaneous type I IFN induction during virus infection.

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Molecular cloning of the breakpoint for 3q27 translocation in B-cell lymphomas and leukemias

Blood ◽

10.1182/blood.v83.1.217.217 ◽

1994 ◽

Vol 83 (1) ◽

pp. 217-222 ◽

Cited By ~ 60

Author(s):

T Miki ◽

N Kawamata ◽

A Arai ◽

K Ohashi ◽

Y Nakamura ◽

...

Keyword(s):

B Cell ◽

Cell Line ◽

Established Cell Line ◽

Chain Gene ◽

Light Chain Gene ◽

Normal Tissues ◽

A Cell ◽

Established Cell ◽

Hematopoietic Cell Lines

Abstract Reciprocal exchanges between chromosomal region 3q27 and three loci of the Ig genes have been reported in cases of B-cell type non-Hodgkin's lymphoma. We have cloned a region containing a breakpoint junction of 3q27 from a cell line established from a patient with Burkitt's lymphoma carrying t(3;22)(q27;q11). The region cloned was shown to contain an Ig lambda light chain gene fused to a gene on chromosome 3q27. This finding was subsequently confirmed by fluorescence in situ hybridization. Extra nucleotides were present at the joining site. The heptamer-like and nonamer-like sequences separated by an intervening 24 bp were present in the region corresponding to the breakpoint of 3q27, suggesting that a misrecombination in Ig gene rearrangement may be involved in the translocation. Southern blot analysis with a 3q27- specific probe showed rearrangements in three additional patients with B-cell malignancies with the t(3;14)(q27;q32). The breakpoints of all four cases clustered within a limited 3-kb region on chromosome 3q27. The region of 3q27 involved in the translocation was designated as the BCL5 locus. The transcripts from the BCL5 locus were detected in normal tissues and hematopoietic cell lines, and the increased expression of transcript of aberrant size was detected in the established cell line carrying t(3;22). These observations suggest that a gene located at 3q27 is involved in the translocation and that its deregulation plays a role in the malignant transformation of B cells.

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Chemical Synthesis, Characterisation, and Biocompatibility of Nanometre Scale Porous Anodic Aluminium Oxide Membranes for Use as a Cell Culture Substrate for the Vero Cell Line: A Preliminary Study

BioMed Research International ◽

10.1155/2014/238762 ◽

2014 ◽

Vol 2014 ◽

pp. 1-10 ◽

Cited By ~ 6

Author(s):

Gérrard Eddy Jai Poinern ◽

Xuan Thi Le ◽

Mark O'Dea ◽

Thomas Becker ◽

Derek Fawcett

Keyword(s):

Cell Culture ◽

Vero Cell ◽

Cell Line ◽

Aluminium Oxide ◽

Cellular Interaction ◽

African Green Monkey Kidney ◽

Anodic Aluminium Oxide ◽

Vero Cell Line ◽

A Cell ◽

Preliminary Study

In this preliminary study we investigate for the first time the biomedical potential of using porous anodic aluminium oxide (AAO) membranes as a cell substrate for culturing theCercopithecus aethiops(African green monkey) Kidney (Vero) epithelial cell line. One advantage of using the inorganic AAO membrane is the presence of nanometre scale pore channels that allow the exchange of molecules and nutrients across the membrane. The size of the pore channels can be preselected by adjusting the controlling parameters of a temperature controlled two-step anodization process. The cellular interaction and response of the Vero cell line with an in-house synthesised AAO membrane, a commercially available membrane, and a glass control were assessed by investigating cell adhesion, morphology, and proliferation over a 72 h period. The number of viable cells proliferating over the respective membrane surfaces revealed that the locally produced in-house AAO membrane had cells numbers similar to the glass control. The study revealed evidence of focal adhesion sites over the surface of the nanoporous membranes and the penetration of cellular extensions into the pore structure as well. The outcome of the study has revealed that nanometre scale porous AAO membranes have the potential to become practical cell culture scaffold substrates with the capability to enhance adhesion and proliferation of Vero cells.

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Generation and Characterization of a Mammalian Cell Line Continuously Expressing Japanese Encephalitis Virus Subviral Particles

Journal of Virology ◽

10.1128/jvi.75.5.2204-2212.2001 ◽

2001 ◽

Vol 75 (5) ◽

pp. 2204-2212 ◽

Cited By ~ 76

Author(s):

Eiji Konishi ◽

Atsuko Fujii ◽

Peter W. Mason

Keyword(s):

Japanese Encephalitis Virus ◽

Cell Line ◽

Cell Lines ◽

Japanese Encephalitis ◽

Current Source ◽

Culture Fluid ◽

Biochemical Properties ◽

Encephalitis Virus ◽

F Cells ◽

A Cell

ABSTRACT We have generated a cell line (F cells) producing a secreted form of Japanese encephalitis virus (JEV) subviral particle (extracellular particles [EPs]) that contains the JEV envelope glycoprotein (E) and a precursor (prM) of the virion membrane protein (M). The F cells were engineered to synthesize these JEV products from a cDNA encoding a mutated (furin proteinase resistant) form of prM, since stable cell lines expressing E and the authentic form of prM could not be obtained, due (in part) to the cell-fusing ability of EPs containing E and M. Our biochemical alteration of the prM protein was critical for the successful production of EP-producing cell lines. EPs produced by F cells share the biochemical properties of empty viral particles produced by JEV-infected cells, except that the F-cell EPs lack hemagglutinating activity and M. F-cell EPs were recognized by a panel of monoclonal antibodies to E, and EPs were shown to be useful as vaccine candidates in mice and as diagnostic reagents in evaluating human immune responses to JE vaccination. The amounts of E antigen released into the culture fluid of F cells were similar to those found in virion fractions of JEV-infected cell culture fluids or JEV-infected weanling mouse brains (the current source of antigen used to produce human vaccines for JE). Thus, the F-cell line would appear to be a useful source of antigen for JE vaccines and diagnostics.

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Aberrant Type C Virus Production in a Cell Line Derived from Balb/3T3

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094693 ◽

1972 ◽

Vol 30 ◽

pp. 286-287

Author(s):

N. Savage ◽

A. Hackett

Keyword(s):

Electron Microscopy ◽

Cell Line ◽

Leukemia Virus ◽

Morphological Characteristics ◽

Virus Production ◽

Oncogenic Viruses ◽

Endogenous Virus ◽

Virus Particles ◽

Moloney Leukemia Virus ◽

A Cell

A cell line, UC1-B, which was derived from Balb/3T3 cells, maintains the same morphological characteristics of the non-transformed parental culture, and shows no evidence of spontaneous virus production. Survey by electron microscopy shows that the cell line consists of spindle-shaped cells with no unusual features and no endogenous virus particles.UC1-B cells respond to Moloney leukemia virus (MLV) infection by a change in morphology and growth pattern which is typical of cells transformed by sarcoma virus. Electron microscopy shows that the cells are now variable in shape (rounded, rhomboid, and spindle), and each cell type has some microvilli. Virtually all (90%) of the cells show virus particles developing at the cell surface and within the cytoplasm. Maturing viruses, typical of the oncogenic viruses, are found along with atypical tubular forms in the same cell.

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