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

• Published in: STAR protocols Vol. 4; no. 2; p. 102343
• Main Authors: Bertzbach, Luca D., Kohn, Marina, You, Yu, Kossak, Lisa, Sabsabi, Mohammad A., Kheimar, Ahmed, Härtle, Sonja, Kaufer, Benedikt B.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.06.2023; Elsevier
• Subjects: Cell Biology; Cell isolation; Cell-based Assays; Microbiology; Protocol; Microbiology; Cell isolation; Cell-based Assays; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102343

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 [Display omitted] •In vitro infection system for primary target cells of MDV•MDV infection of primary lymphocytes and lymphocyte cell lines•Facilitates evaluation of MDV lytic replication, latency, and transformation•Reduces need for animal experiments to investigate this oncogenic herpesvirus Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.