Fachbereich Veterinärmedizin



    In vitro model for lytic replication, latency, and transformation of an oncogenic alphaherpesvirus (2015)

    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Schermuly, Julia
    Greco, Annachiara (WE 5)
    Härtle, Sonja
    Osterrieder, Nikolaus (WE 5)
    Kaufer, Benedikt B (WE 5)
    Kaspers, Bernd
    Proceedings of the National Academy of Sciences of the United States of America; 112(23) — S. 7279–7284
    ISSN: 0027-8424
    DOI: 10.1073/pnas.1424420112
    Pubmed: 26039998
    Institut für Virologie

    Robert-von-Ostertag-Str. 7-13
    Gebäude 35
    14163 Berlin
    Tel. +49 30 838 51833 Fax. +49 30 838 451847

    Abstract / Zusammenfassung

    Marek's disease virus (MDV) is an alphaherpesvirus that causes deadly T-cell lymphomas in chickens and serves as a natural small animal model for virus-induced tumor formation. In vivo, MDV lytically replicates in B cells that transfer the virus to T cells in which the virus establishes latency. MDV also malignantly transforms CD4+ T cells with a T(reg) signature, ultimately resulting in deadly lymphomas. No in vitro infection system for primary target cells of MDV has been available due to the short-lived nature of these cells in culture. Recently, we characterized cytokines and monoclonal antibodies that promote survival of cultured chicken B and T cells. We used these survival stimuli to establish a culture system that allows efficient infection of B and T cells with MDV. We were able to productively infect with MDV B cells isolated from spleen, bursa or blood cultured in the presence of soluble CD40L. Virus was readily transferred from infected B to T cells stimulated with an anti-TCRαVβ1 antibody, thus recapitulating the in vivo situation in the culture dish. Infected T cells could then be maintained in culture for at least 90 d in the absence of TCR stimulation, which allowed the establishment of MDV-transformed lymphoblastoid cell lines (LCL). The immortalized cells had a signature comparable to MDV-transformed CD4+ α/β T cells present in tumors. In summary, we have developed a novel in vitro system that precisely reflects the life cycle of an oncogenic herpesivrus in vivo and will allow us to investigate the interaction between virus and target cells in an easily accessible system.