In vitro infection of primary chicken lymphocytes with Marek's disease virus

Luca D Bertzbach; Marina Kohn; Yu You; Lisa Kossak; Mohammad A Sabsabi; Ahmed Kheimar; Sonja Härtle; Benedikt B Kaufer

doi:10.1016/j.xpro.2023.102343

In vitro infection of primary chicken lymphocytes with Marek's disease virus

STAR Protoc. 2023 Jun 3;4(2):102343. doi: 10.1016/j.xpro.2023.102343. Online ahead of print.

Authors

Luca D Bertzbach¹, Marina Kohn², Yu You³, Lisa Kossak³, Mohammad A Sabsabi³, Ahmed Kheimar⁴, Sonja Härtle⁵, Benedikt B Kaufer⁶

Affiliations

¹ Freie Universität Berlin, Institute of Virology, 14163 Berlin, Germany; Leibniz Institute of Virology (LIV), Department of Viral Transformation, 20251 Hamburg, Germany.
² Ludwig-Maximilians-Universität, Department for Veterinary Sciences, 82152 Planegg/Martinsried, Germany.
³ Freie Universität Berlin, Institute of Virology, 14163 Berlin, Germany.
⁴ Freie Universität Berlin, Institute of Virology, 14163 Berlin, Germany; Sohag University, Faculty of Veterinary Medicine, Department of Poultry Diseases, 82524 Sohag, Egypt.
⁵ Ludwig-Maximilians-Universität, Department for Veterinary Sciences, 82152 Planegg/Martinsried, Germany. Electronic address: sonja.haertle@lmu.de.
⁶ Freie Universität Berlin, Institute of Virology, 14163 Berlin, Germany; Freie Universität Berlin, Veterinary Centre for Resistance Research (TZR), 14163 Berlin, Germany. Electronic address: benedikt.kaufer@fu-berlin.de.

Abstract

Marek's disease virus (MDV) is a highly oncogenic alphaherpesvirus that infects immune cells and causes a deadly lymphoproliferative disease in chickens. Cytokines and monoclonal antibodies promote the survival of chicken lymphocytes in vitro. Here, we describe protocols for the isolation, maintenance, and efficient MDV infection of primary chicken lymphocytes and lymphocyte cell lines. This facilitates the investigation of key aspects of the MDV life cycle in the primary target cells of viral replication, latency, genome integration, and reactivation. For complete details on the use and execution of this protocol, please refer to Schermuly et al.,¹ Bertzbach et al. (2019),² and You et al.³ For a comprehensive background on MDV, please see Osterrieder et al.⁴ and Bertzbach et al. (2020).⁵.

Keywords: Cell Biology; Cell isolation; Cell-based Assays; Microbiology.