title: Production of herpes B virus recombinant glycoproteins and evaluation of their diagnostic potential.
abstract: B virus (cercopithecine herpesvirus 1) is the only deadly alphaherpesvirus that is zoonotically transmissible from macaques to humans. The detection of humoral immune responses is the method of choice for the rapid identification of B virus-infected animals. We evaluated the diagnostic potential of recombinant B virus glycoproteins for the detection of immunoglobulin G (IgG) antibodies in monkey and human sera. Glycoproteins B, C, and E and secreted (sgG) and membrane-associated (mgG) segments of glycoprotein G (gG) were expressed in the baculovirus expression system, while gD was expressed in CHO cells. We developed recombinant protein-based IgG enzyme-linked immunosorbent assays (ELISAs) and compared their diagnostic efficacies by using B virus antibody-negative (n = 40) and -positive (n = 75) macaque sera identified by a whole antigen-based ELISA and Western blotting. The diagnostic sensitivities of the gB-, gC-, gD-, and mgG-ELISAs were 100, 97.3, 88.0, and 80.0%, respectively. The specificities of the gB-, gC-, and gD-ELISAs and of the mgG-ELISA were 100 and 97.5%, respectively. In contrast, the sensitivities and specificities of sgG- and gE-ELISAs were low, suggesting that sgG and gE are less effective diagnostic antigens. Sera from nonmacaque monkeys cross-reacted with gB, gC, and gD, and only baboon sera reacted weakly with mgG. Human herpes simplex virus type 1 (HSV-1)- and HSV-2-positive sera pools reacted with gB and gD, whereas sera from B virus-infected individuals reacted with all four antigens. These data indicate that gB, gC, gD, and mgG have a high diagnostic potential for B virus serodiagnosis in macaques, whereas mgG may be a valuable antigen for discrimination between antibodies induced by B virus and those induced by other, closely related alphaherpesviruses, including HSV-1 and -2.
authors.affiliation: Viral Immunology Center, Georgia State University, P.O. Box 4118, Atlanta, GA 30302-4118, USA. biolmp@panther.gsu.edu
authors.forename: Ludmila
authors.initials: L
authors.lastname: Perelygina
authors.forename: Irina
authors.initials: I
authors.lastname: Patrusheva
authors.forename: Soumya
authors.initials: S
authors.lastname: Hombaiah
authors.forename: Holley
authors.initials: H
authors.lastname: Zurkuhlen
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Wildes
authors.forename: Nikolai
authors.initials: N
authors.lastname: Patrushev
authors.forename: Julia
authors.initials: J
authors.lastname: Hilliard
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000821:virology; D006568:Herpesvirus 1, Cercopithecine / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D008992:Monkey Diseases / Q000175:diagnosis / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
chemical_list: D000914:Antibodies, Viral; D006023:Glycoproteins; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins