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Archives of Virology

, Volume 163, Issue 6, pp 1519–1530 | Cite as

Development of a luminescence syncytium induction assay (LuSIA) for easily detecting and quantitatively measuring bovine leukemia virus infection

  • Hirotaka Sato
  • Sonoko Watanuki
  • Hironobu Murakami
  • Reiichiro Sato
  • Hiroshi Ishizaki
  • Yoko Aida
Original Article

Abstract

Bovine leukemia virus (BLV) causes enzootic bovine leukosis and is closely related to the human T cell leukemia virus. Since BLV infection mostly occurs via cell-to-cell transmission, BLV infectivity is generally measured by culturing BLV-infected cells with reporter cells that form syncytia upon BLV infection. However, this method is time-consuming and requires skill. To visualize the infectivity of BLV, we developed a new assay called the luminescence syncytium induction assay (LuSIA) that is based on a new reporter cell line designated CC81-BLU3G. CC81-BLU3G is stably transfected with pBLU3-EGFP, which contains the BLV long terminal repeat U3 region linked to the enhanced-green fluorescence protein (EGFP) gene. CC81-BLU3G expresses the EGFP in response to BLV Tax expression specifically, and forms fluorescing syncytia when transfected with an infectious BLV plasmid or when cultured with BLV-infected cells. Compared to the conventional assay, LuSIA was more specific and detected cattle samples with low proviral loads. The fluorescing syncytia was easily detected by eye and automated scanning and LuSIA counts correlated strongly with the proviral load of infected cattle (R2 = 0.8942).

Notes

Acknowledgements

The authors thank Ms. Yoshiko Sakuma for providing technical support and Dr. Meripet Polat for constructing the pME18neo/BLV Tax-FLAG plasmid. We also thank all members of the virus infectious disease unit of RIKEN, Grazing Animal Unit, Institute of Livestock and Grassland Science and Nasu Operation Unit, Technical Support Center, NARO for providing advice and samples. We are grateful to the Support Unit, Bio-material Analysis, RIKEN BSI Research Resources Center for helping with the sequence analysis.

Funding information

This study was supported by Grants-in-Aid for Scientific Research (A) and Young Scientists (B) from the Japan Society for the Promotion of Science (JSPS) [Grant Nos. JP16H02590 and JP17K18356], and by grants from the Project of the NARO Bio-oriented Technology Research Advancement Institution (the special scheme regarding regional development strategies) [Grant No. 16817983].

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interests to declare.

Ethics statement

All animal experiments were conducted in accordance with the Guidelines for Laboratory Animal Welfare and Animal Experiment Control that were set out by the Institute of Livestock and Grassland Science, NARO (permit numbers: 1711B082) and by the School of Veterinary Medicine of Azabu University (permit numbers: 161121-2).

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Hirotaka Sato
    • 1
    • 2
  • Sonoko Watanuki
    • 1
    • 2
    • 3
  • Hironobu Murakami
    • 4
  • Reiichiro Sato
    • 5
  • Hiroshi Ishizaki
    • 6
  • Yoko Aida
    • 1
    • 2
  1. 1.Virus Infectious Diseases UnitRIKENWakoJapan
  2. 2.Nano Medical Engineering LaboratoryRIKENWakoJapan
  3. 3.Laboratory of Global Animal Resource Science, Graduate School of Agricultural and Life ScienceThe University of TokyoTokyoJapan
  4. 4.Laboratory of Animal Health II, School of Veterinary MedicineAzabu UniversitySagamiharaJapan
  5. 5.Laboratory of Farm Animal Internal Medicine, School of Veterinary MedicineAzabu UniversitySagamiharaJapan
  6. 6.Grazing Animal Unit, Division of Grassland FarmingInstitute of Livestock and Grassland Sciences, NARONasushiobaraJapan

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