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Title: SEROLOGICAL CHARACTERISTICS OF A MEMBRANE GLYCOPROTEIN GP82 OF MAREK'S DISEASE VIRUS

Author
item WU, PING - MICHIGAN STATE UNIVERSITY
item Lee, Lucy
item REED, WILLIE - MICHIGAN STATE UNIVERSITY

Submitted to: Avian Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/30/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary: Marek's disease virus (MDV) is a herpes virus and causes Marek's disease in chickens resulting in tumor induction. The research goal was to identify ways to prevent disease and tumor induction. One of the approaches used was to find MDV genes produce products which can be used as a component of vaccine to protect against disease. This paper describes the development of a monoclonal antibody to study the characteristics of a newly-identified MDV protein. This monoclonal antibody will be a useful reagent to identify this protein in MDV-infected cells as well as in evaluating recombinant viruses containing this protein for vaccine development.

Technical Abstract: Marek's disease virus (MDV) is a cell-associated herpesvirus of gallinaceous fowl. Glycoproteins of herpesvirus have been reported to be important in stimulating humoral and cell-mediated immune responses against virus infection. At least nine glycoproteins of MDV have been identified and glycoprotein B (gB) has been reported to produce protective immunity. In addition, other proteins expressed in cell membrane may also be important for engendering immunity. We developed three monoclonal antibodies and rabbit polyclonal antibodies specific for MDV membrane protein gp82 in an attempt to determine whether the protein is involved in immune responses. Three Mabs (Mab2.1, 3.7, and 5.7) were produced by using TrpEgp82 fusion protein as the immunogen which was expressed by recombinant pATH expression vector pATHgp82 in E. coli. Indirect immunofluorescence assay (IFA), immunoprecipitation, and Western blot analysis are used to determine the characteristics of gp82. All of the Mabs reacted with gp82 protein irrespective of its conformation. The gp82 protein was predominantly anchored on the surface membrane of chicken embryo fibroblast (CEF) infected with MDV-1. There was no detectable immunofluorescence staining in the cytoplasm or nuclear areas in the MDV-1 infected CEFs. These results indicated gp82 is a membrane protein. There are strain differences in expression morphology of gp82. The distribution pattern of immunofluorescence staining varied between CEFs infected with low and high passage MDV11 strains. In HVT (MDV-3) and SB-1 (MDV-3) infected CEFs, there was no gp82 product detected by immunoprecipation, Western blot analysis, or IFA using both monoclonal and polyclonal antibodies.