Mammals Preferred: Reassortment of Batai and Bunyamwera orthobunyavirus Occurs in Mammalian but Not Insect Cells

Reassortment is a viral genome-segment recomposition known for many viruses, including the orthobunyaviruses. The co-infection of a host cell with two viruses of the same serogroup, such as the Bunyamwera orthobunyavirus and the Batai orthobunyavirus, can give rise to novel viruses. One example is the Ngari virus, which has caused major outbreaks of human infections in Central Africa. This study aimed to investigate the potential for reassortment of Bunyamwera orthobunyavirus and the Batai orthobunyavirus during co-infection studies and the replication properties of the reassortants in different mammalian and insect cell lines. In the co-infection studies, a Ngari-like virus reassortant and a novel reassortant virus, the Batunya virus, arose in BHK-21 cells (Mesocricetus auratus). In contrast, no reassortment was observed in the examined insect cells from Aedes aegypti (Aag2) and Aedes albopictus (U4.4 and C6/36). The growth kinetic experiments show that both reassortants are replicated to higher titers in some mammalian cell lines than the parental viruses but show impaired growth in insect cell lines.

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Recovery of infective juveniles of the entomopathogenic nematode Steinernema carpocapsae via factors produced by insect cells and symbiotic bacteria

Nematology ◽

10.1163/138855409x12465362560719 ◽

2009 ◽

Vol 11 (4) ◽

pp. 611-618 ◽

Cited By ~ 2

Author(s):

Shingo Kikuta ◽

Takashi Kiuchi ◽

Fugaku Aoki ◽

Masao Nagata

Keyword(s):

Cell Lines ◽

Insect Cell ◽

Mammalian Cells ◽

Insect Cells ◽

Entomopathogenic Nematode ◽

Insect Cell Line ◽

Fruit Fly ◽

Steinernema Carpocapsae ◽

Symbiotic Bacteria ◽

Insect Cell Lines

Abstract Entomopathogenic nematodes, Steinernema carpocapsae, show 'recovery' from the dauer form as infective juveniles (IJ) up to fourth-stage juveniles when host invasion occurs. This recovery also occurs within an insect cell line culturing system. Here we addressed the factor(s) that induce recovery. When IJ were exposed to cell medium obtained from the cultivation of Sf9 cell lines derived from armyworms (Spodoptera frugiperda), approximately 50% of IJ recovered after 4 h. By 16 h, 90% of the IJ had undergone recovery. Other insect cell lines such as silkworm (Bombyx mori)-derived BmN cells and fruit fly (Drosophila melanogaster)-derived S2 cells also secreted the recovery inducing factor(s). By contrast, mammalian cells (NIH/3T3 and HeLa) had no effect on nematode recovery. Our data also suggest that symbiotic bacteria are involved in IJ recovery; axenic IJ did not recover in the cell-cultured medium. When symbiotic bacteria isolated from IJ were propagated within the cell-cultured medium, the supernatant gained recovery-inducing activity against axenic IJ. From these results, we conclude that IJ recovery in S. carpocapsae is induced by multiple factor(s) secreted from insect cells and symbiont bacteria.

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The thyrotrophin hormone receptor of Graves' disease: overexpression of the extracellular domain in insect cells using recombinant baculovirus, immunoaffinity purification and analysis of autoantibody binding

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0100127 ◽

1993 ◽

Vol 10 (2) ◽

pp. 127-142 ◽

Cited By ~ 47

Author(s):

G C Huang ◽

M J Page ◽

L B Nicholson ◽

K S Collison ◽

A M McGregor ◽

...

Keyword(s):

Monoclonal Antibody ◽

Cell Lines ◽

Insect Cell ◽

Insect Cells ◽

Recombinant Baculovirus ◽

Receptor Expression ◽

Graves Disease ◽

Truncated Receptor ◽

Immunoaffinity Purification ◽

Insect Cell Lines

ABSTRACT Since the cloning of the TSH receptor (TSH-R), the target autoantigen of Graves' disease, the receptor has been expressed in a variety of eukaryotic cells to obtain a functional molecule. Despite this success, the levels of receptor expression have been marginally higher than the extremely low levels found in thyroid cells, preventing any progress on the purification of the molecule. In this study, the large extracellular region of the TSH-R, without the membrane spanning segments, has been expressed in insect cells using recombinant baculovirus to generate substantial quantities of the receptor protein. A monoclonal antibody previously generated to a bacterial TSH-R fusion protein was used to characterize and monitor the expression of the truncated receptor in insect cells. Two polypeptides of 63 and 49 kDa were recognized as the components of the truncated recombinant receptor. The 63 kDa protein was shown to be the glycosylated form of the smaller, 49 kDa, component. Expression in different insect cell lines showed that an increase in expression of approximately tenfold was apparent in High Five cells when compared with Sf21 cells. Very small quantities of the truncated receptor were secreted by the three insect cell lines examined, with the majority of the molecule being retained within the cells. Immunoaffinity purification of milligram quantities of the truncated receptor was achieved using the monoclonal antibody. The availability of the purified TSH-R has allowed the establishment of an enzymelinked immunosorbent assay to measure autoantibodies in the sera of patients with Graves' disease. Although the truncated receptor interacts with autoantibodies, our results show that it does not bind TSH and differs in this respect from other glycoprotein hormone receptors.

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