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Journal of Virology, June 2001, p. 5159-5173, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.5159-5173.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Marek's Disease Virus (MDV) Encodes an Interleukin-8 Homolog (vIL-8): Characterization of the vIL-8 Protein and a vIL-8 Deletion Mutant MDV†

Mark S. Parcells,1,* Su-Fang Lin,2 Robert L. Dienglewicz,1 Vladimir Majerciak,3 Dan R. Robinson,2 Hua-Chien Chen,2 Zining Wu,4 George R. Dubyak,5 Peter Brunovskis,2 Henry D. Hunt,4 Lucy F. Lee,6 and Hsing-Jien Kung2,Dagger

Center of Excellence for Poultry Science, Department of Poultry Science, University of Arkansas, Fayetteville, Arkansas 727011; Department of Molecular Biology & Microbiology2 and Department of Physiology & Biophysics,5 School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106; Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovak Republic3; Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 037554; and USDA-ARS, Avian Disease and Oncology Laboratory, East Lansing, Michigan 488236

Received 16 November 2000/Accepted 9 March 2001

Chemokines induce chemotaxis, cell migration, and inflammatory responses. We report the identification of an interleukin-8 (IL-8) homolog, termed vIL-8, encoded within the genome of Marek's disease virus (MDV). The 134-amino-acid vIL-8 shares closest homology to mammalian and avian IL-8, molecules representing the prototype CXC chemokine. The gene for vIL-8 consists of three exons which map to the BamHI-L fragment within the repeats flanking the unique long region of the MDV genome. A 0.7-kb transcript encoding vIL-8 was detected in an n-butyrate-treated, MDV-transformed T-lymphoblastoid cell line, MSB-1. This induction is essentially abolished by cycloheximide and herpesvirus DNA polymerase inhibitor phosphonoacetate, indicating that vIL-8 is expressed with true late (gamma 2) kinetics. Baculovirus-expressed vIL-8 was found to be secreted into the medium and shown to be functional as a chemoattractant for chicken peripheral blood mononuclear cells but not for heterophils. To characterize the function of vIL-8 with respect to MDV infection in vivo, a recombinant MDV was constructed with a deletion of all three exons and a soluble-modified green fluorescent protein (smGFP) expression cassette inserted at the site of deletion. In two in vivo experiments, the vIL-8 deletion mutant (RB1BvIL-8Delta smGFP) showed a decreased level of lytic infection in comparison to its parent virus, an equal-passage-level parent virus, and to another recombinant MDV containing the insertion of a GFP expression cassette at the nonessential US2 gene. RB1BvIL-8Delta smGFP retained oncogenicity, albeit at a greatly reduced level. Nonetheless, we have been able to establish a lymphoblastoid cell line from an RB1BvIL-8Delta smGFP-induced ovarian lymphoma (MDCC-UA20) and verify the presence of a latent MDV genome lacking vIL-8. Taken together, these data describe the identification and characterization of a chemokine homolog encoded within the MDV genome that is dispensable for transformation but may affect the level of MDV in vivo lytic infection.

* Corresponding author. Mailing address: O-404, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR 72701. Phone: (501) 575-5494. Fax: (501) 575-7139. E-mail: parcells{at}uark.edu.

dagger Arkansas Agricultural Experiment Station manuscript no. 00108.

Dagger Present address: UC Davis Cancer Center, Sacramento, CA 95817.

Journal of Virology, June 2001, p. 5159-5173, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.5159-5173.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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