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Novel serological tools for detection of Thottapalayam virus, a Soricomorpha-borne hantavirus

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Abstract

We developed serological tools for the detection of hantavirus-specific antibodies and hantavirus antigens in shrews. The work was focussed to generate Thottapalayam virus (TPMV)-specific monoclonal antibodies (mAbs) and anti-shrew immunoglobulin G (IgG) antibodies. The mAbs against TPMV nucleocapsid (N) protein were produced after immunization of BALB/c mice with recombinant TPMV N proteins expressed in Escherichia coli, baculovirus and Saccharomyces cerevisiae-mediated expression systems. In total, six TPMV N-protein-specific mAbs were generated that showed a characteristic fluorescent pattern in indirect immunofluorescence assay (IFA) using TPMV-infected Vero cells. Out of the six mAbs tested, five showed no cross-reaction to rodent-associated hantaviruses (Hantaan, Seoul, Puumala, Tula, Dobrava-Belgrade and Sin Nombre viruses) in IFA and enzyme-linked immunosorbent assay (ELISA), although one mAb reacted to Sin Nombre virus in IFA. None of the mAbs cross-reacted with an amino-terminal segment of the shrew-borne Asama virus N protein. Anti-shrew-IgG sera were prepared after immunization of rabbits and BALB/c-mice with protein-G-purified shrew IgG. TPMV-N-protein-specific sera were raised by immunisation of Asian house shrews (Suncus murinus) with purified yeast-expressed TPMV N protein. Using these tools, an indirect ELISA was developed to detect TPMV-N-protein-specific antibodies in the sera of shrews. Using an established serological assay, high TPMV N protein specific antibody titres were measured in the sera of TPMV-N-protein-immunized and experimentally TPMV-infected shrews, whereas no cross-reactivity to other hantavirus N proteins was found. Therefore, the generated mAbs and the established ELISA system represent useful serological tools to detect TPMV, TPMV-related virus antigens or hantavirus-specific antibodies in hantavirus-infected shrews.

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Acknowledgments

The authors kindly acknowledge the support of Kathrin Heidemanns, Peter Giere and Swetlana Siniza and the critical reading of Daniel Balkema. The mAbs 2C6, 7A5, 5E11, 5C5, 7G2, 4H3, and 4C3, 2E12, 5A3, 1C12 were kindly provided by Aurelija Zvirbliene (Vilnius) and Åke Lundkvist (Stockholm). This study was supported in part by the Program of Founding Research Centers for Emerging and Reemerging Infectious Diseases, Ministry of Education, Culture, Sports, Science and Technology, Japan. This work was also supported in part by a grant from the Global COE program (Establishment of International Collaboration Centers for Zoonosis Control) and by Grants-in-Aid for Research on Emerging and Re-emerging Infectious Diseases from the Ministry of Health, Labour and Welfare. In addition, the study was partially funded by EU grant FP7-261504 EDENext and is catalogued by the EDENext Steering Committee as EDENext018 (http://www.edenext.eu). The contents of this publication are the sole responsibility of the authors and do not necessarily reflect the views of the European Commission.

Ethical statement

Shrews, laboratory mice and rabbits were handled according to the Laboratory Animal Control Guidelines of the Hokkaido University Animal Research Committee in Japan and EU Council Directive 86/609/EEC for the protection of animals used for experiments in Germany.

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Authors and Affiliations

  1. Friedrich-Loeffler-Institut, Institute for Novel and Emerging Infectious Diseases, Greifswald-Insel Riems, Germany

    Mathias Schlegel, Marc Mertens, Martin H. Groschup & Rainer G. Ulrich

  2. Department of Laboratory Animal Science, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Japan

    Erdenesaikhan Tegshduuren & Chiaki Ishihara

  3. Department of Microbiology, Graduate School of Medicine, Hokkaido University, Kita-ku, Kita-18, Nishi-7, Sapporo, Hokkaido, 060-8638, Japan

    Kumiko Yoshimatsu, Rika Endo, Kenta Shimizu, Takaaki Koma, Shumpei Yasuda & Jiro Arikawa

  4. Vilnius University, Institute of Biotechnology, Vilnius, Lithuania

    Rasa Petraityte & Kestutis Sasnauskas

  5. Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, Greifswald, Insel Riems, Germany

    Bärbel Hammerschmidt

  6. DSP Process Development, Glycotope GmbH, Berlin, Germany

    Robert Friedrich

  7. National Institute of Infectious Diseases, Tokyo, Japan

    Satoru Arai

  8. Institute of Immunology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Insel Riems, Greifswald, Germany

    Bernd Köllner

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  1. Mathias Schlegel
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  2. Erdenesaikhan Tegshduuren
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  3. Kumiko Yoshimatsu
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Correspondence to Jiro Arikawa or Bernd Köllner.

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M. Schlegel, E. Tegshduuren, J. Arikawa and B. Köllner contributed equally to this article.

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Schlegel, M., Tegshduuren, E., Yoshimatsu, K. et al. Novel serological tools for detection of Thottapalayam virus, a Soricomorpha-borne hantavirus. Arch Virol 157, 2179–2187 (2012). https://doi.org/10.1007/s00705-012-1405-9

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