title: In vitro model for lytic replication, latency, and transformation of an oncogenic alphaherpesvirus.
abstract: 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.
authors.affiliation: Veterinärwissenschaftliches Department, Institut für Tierphysiologie, Ludwig-Maximilians-Universität München, 80539 Munich, Germany;
authors.forename: Julia
authors.initials: J
authors.lastname: Schermuly
authors.affiliation: Institut für Virologie, Freie Universität Berlin, 14163 Berlin, Germany.
authors.forename: Annachiara
authors.initials: A
authors.lastname: Greco
authors.affiliation: Veterinärwissenschaftliches Department, Institut für Tierphysiologie, Ludwig-Maximilians-Universität München, 80539 Munich, Germany;
authors.forename: Sonja
authors.initials: S
authors.lastname: Härtle
authors.affiliation: Institut für Virologie, Freie Universität Berlin, 14163 Berlin, Germany no.34@fu-berlin.de b.kaufer@fu-berlin.de kaspers@lmu.de.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Osterrieder
authors.affiliation: Institut für Virologie, Freie Universität Berlin, 14163 Berlin, Germany no.34@fu-berlin.de b.kaufer@fu-berlin.de kaspers@lmu.de.
authors.forename: Benedikt B
authors.initials: BB
authors.lastname: Kaufer
authors.affiliation: Veterinärwissenschaftliches Department, Institut für Tierphysiologie, Ludwig-Maximilians-Universität München, 80539 Munich, Germany; no.34@fu-berlin.de b.kaufer@fu-berlin.de kaspers@lmu.de.
authors.forename: Bernd
authors.initials: B
authors.lastname: Kaspers
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000821:virology; D002469:Cell Separation; D002471:Cell Transformation, Neoplastic; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002645:Chickens; D005434:Flow Cytometry; D005814:Genes, Viral; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D066298:In Vitro Techniques; D025922:Mardivirus / Q000235:genetics / Q000502:physiology*; D013601:T-Lymphocytes / Q000821:virology; D017735:Virus Latency; D014779:Virus Replication
keywords: B cells; Marek’s disease virus; T cells; genomic integration; lymphomagenesis