A Cell Culture System to Investigate Marek’s Disease Virus Integration into Host Chromosomes

You, Yu; Vychodil, Tereza; Aimola, Giulia; Previdelli, Renato L.; Göbel, Thomas W.; Bertzbach, Luca D.; Kaufer, Benedikt B.

doi:10.3390/microorganisms9122489

Open AccessCommunication

A Cell Culture System to Investigate Marek’s Disease Virus Integration into Host Chromosomes

by

Yu You

^1,†,

Tereza Vychodil

^1,†,

Giulia Aimola

¹,

Renato L. Previdelli

^1,2,

Thomas W. Göbel

³,

Luca D. Bertzbach

^1,4,*

and

Benedikt B. Kaufer

^1,*

¹

Institute of Virology, Freie Universität Berlin, 14163 Berlin, Germany

²

Department of Comparative Biomedical Sciences, Royal Veterinary College, London NW1 0TU, UK

³

Institute for Animal Physiology, Department of Veterinary Sciences, Ludwig Maximilian University Munich, 80539 Munich, Germany

⁴

Department of Viral Transformation, Leibniz Institute for Experimental Virology (HPI), 20251 Hamburg, Germany

^*

Authors to whom correspondence should be addressed.

^†

Y.Y. and T.V. contributed equally to this work.

Microorganisms 2021, 9(12), 2489; https://doi.org/10.3390/microorganisms9122489

Submission received: 18 October 2021 / Revised: 25 November 2021 / Accepted: 29 November 2021 / Published: 1 December 2021

(This article belongs to the Special Issue Marek’s Disease Virus)

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Abstract

Marek’s disease virus (MDV) is a highly oncogenic alphaherpesvirus that causes a devastating neoplastic disease in chickens. MDV has been shown to integrate its genome into the telomeres of latently infected and tumor cells, which is crucial for efficient tumor formation. Telomeric repeat arrays present at the ends of the MDV genome facilitate this integration into host telomeres; however, the integration mechanism remains poorly understood. Until now, MDV integration could only be investigated qualitatively upon infection of chickens. To shed further light on the integration mechanism, we established a quantitative integration assay using chicken T cell lines, the target cells for MDV latency and transformation. We optimized the infection conditions and assessed the establishment of latency in these T cells. The MDV genome was efficiently maintained over time, and integration was confirmed in these cells by fluorescence in situ hybridization (FISH). To assess the role of the two distinct viral telomeric repeat arrays in the integration process, we tested various knockout mutants in our in vitro integration assay. Efficient genome maintenance and integration was thereby dependent on the presence of the telomeric repeat arrays in the virus genome. Taken together, we developed and validated a novel in vitro integration assay that will shed light on the integration mechanism of this highly oncogenic virus into host telomeres.

Keywords: MDV; herpesvirus; telomere integration; in vitro assay; telomeric repeats; fluorescence in situ hybridization; genome maintenance; latency; viral transformation

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MDPI and ACS Style

You, Y.; Vychodil, T.; Aimola, G.; Previdelli, R.L.; Göbel, T.W.; Bertzbach, L.D.; Kaufer, B.B. A Cell Culture System to Investigate Marek’s Disease Virus Integration into Host Chromosomes. Microorganisms 2021, 9, 2489. https://doi.org/10.3390/microorganisms9122489

AMA Style

You Y, Vychodil T, Aimola G, Previdelli RL, Göbel TW, Bertzbach LD, Kaufer BB. A Cell Culture System to Investigate Marek’s Disease Virus Integration into Host Chromosomes. Microorganisms. 2021; 9(12):2489. https://doi.org/10.3390/microorganisms9122489

Chicago/Turabian Style

You, Yu, Tereza Vychodil, Giulia Aimola, Renato L. Previdelli, Thomas W. Göbel, Luca D. Bertzbach, and Benedikt B. Kaufer. 2021. "A Cell Culture System to Investigate Marek’s Disease Virus Integration into Host Chromosomes" Microorganisms 9, no. 12: 2489. https://doi.org/10.3390/microorganisms9122489

APA Style

You, Y., Vychodil, T., Aimola, G., Previdelli, R. L., Göbel, T. W., Bertzbach, L. D., & Kaufer, B. B. (2021). A Cell Culture System to Investigate Marek’s Disease Virus Integration into Host Chromosomes. Microorganisms, 9(12), 2489. https://doi.org/10.3390/microorganisms9122489

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

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A Cell Culture System to Investigate Marek’s Disease Virus Integration into Host Chromosomes

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