Bovine natural killer-like cell responses against cell lines expressing recombinant bovine herpesvirus type 1 glycoproteins.
The murine retrovirus shuttle vector pZipNeo SV(X)1 was used to construct plasmids encoding the three major surface glycoproteins of bovine herpesvirus-1 (BHV-1). Each plasmid was transfected into D17, a canine osteosarcoma cell line sensitive to lysis by bovine NK-like cells when infected with BHV-1. After selection in G418 sulfate, cell lines expressing the recombinant gene products were sorted by flow microfluorimetry, radioimmunoprecipitated, and analyzed by SDS-PAGE for fidelity as compared to the native viral glycoproteins. Two of the three genetically engineered cell lines (gI and gIV) could successfully serve as targets to detect bovine NK-like cytolysis. These findings support and extend a previous study from our laboratory indicating a role for BHV-1 glycoproteins in the cytolytic response by bovine NK-like cells. Additionally, this study demonstrated that individual proteins are recognized by these effector cells, and that recombinant glycoproteins can direct cytolytic activity in the absence of host cell infection-associated proteins. This is the first known report of Ag directed cytotoxicity by bovine null (non-B, non-T) cells.
Palmer
L D
LD
Department of Veterinary Science, University of Wisconsin-Madison 53706.
Leary
T P
TP
Wilson
D M
DM
Splitter
G A
GA
D000818:Animals; D002417:Cattle; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D007242:Herpesvirus 1, Bovine / Q000276:immunology*; D007694:Killer Cells, Natural / Q000276:immunology*; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology*
D011994:Recombinant Proteins; D014764:Viral Proteins; C046998:bovine herpesvirus type-1 glycoproteins